2015—2020

Life Science Training from Industry Experts

B I O T E C H P R I M E R ’ S

W E E K LY
C O M P E N D I U M

Life Science Training From Industry Experts

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 Life Science Training from Industry Experts

Dear Reader,
The biopharma industry is moving at lighting speed and it can be a challenge to keep
pace. Here at Biotech Primer we spend hours each week researching, writing, and editing
original content for the Biotech Primer WEEKLY with one goal in mind: to help everyone
better understand the latest science and technology driving today’s healthcare industry.
This interactive eBook is a collection of Biotech Primer WEEKLYs from the past five years.
The WEEKLY is a one-page (sometimes two), easy-to-read newsletter delivered each
Thursday directly to your inbox. If you are not registered to receive the Biotech Primer
WEEKLY go to WEEKLY.BiotechPrimer.com and subscribe.
Thank you for downloading the Biotech Primer WEEKLY Compendium 2015-2020.

Stacey Hawkins
Stacey Hawkins
CEO, Biotech Primer
[email protected]

Biotech Primer develops and delivers customized training in six areas:

• Biotechnology for the Non-Scientist • Manufacturing
• Drug Development • The Business of Biotech
• Medical Devices and Diagnostics • Biosafety

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understanding today’s advanced therapeutics
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CONTENTS Life Science Training from Industry Experts

1 HOPE ON THE HORIZON FOR HUNTINGTON’S 70 Rare Disease Focus: PKU

4 THE INFINITESIMAL ENGINES OF BIOLOGICS 72 Red Blood Cells: Ready For Double-Duty?
6 EX-CELL-ENT RESEARCH PARTNERS 74 Gluten: Protein of Doom
8 SEARCH AND DESTROY: CHIMERIC 76 Breaking Bad With SCLC & NSCLC
ANTIGEN RECEPTOR T-CELLS 78 Phage Therapy: New Hope For
10 BLUEPRINT OF THE CENTURY: DNA Antibiotic-Resistant Bacteria
13 SPOTLIGHT ON MACULAR DEGENERATION 80 The Immune System: Friend & Sometimes Foe
15 GETTING THE JUMP ON ALZHEIMER’S DISEASE 82 Amazing Antibodies Part One: Bispecifics
17 EXOSOMES: TINY VESICLES WITH BIG POTENTIAL & Antibody-Drug Conjugates

19 NEW MUSCLE IN THE FIGHT AGAINST DMD 84 Amazing Antibodies Part Two: Enlightened & Nano

21 CONTROLLING A KILLER, ONE STEP AT A TIME 86 Pills, Peptides, & Proteins

23 WORLD AIDS DAY: UNDERSTANDING HIV 88 Epigenetics: Writing, Reading, & Erasing

25 A BREATH OF FRESH AIR FOR 91 Breath Biopsies

CYSTIC FIBROSIS PATIENTS 93 Fountains of Healthy Old Age?
27 VAMPIRES, ZOMBIES, PUMPKINS, & BIOTECH 95 Hacking The Fountain Of Youth
29 NEW PEANUT ALLERGY TREATMENT 97 Attack Of The Tregs!!
ON THE HORIZON 99 Autophagy: The Incredible, Edible Cell?
31 BREATHE EASY: HYPOXIA-INDUCIBLE FACTOR (HIF) 101 Immune Checkpoint Inhibitors
33 NEOANTIGEN CANCER VACCINES 103 Influenza: So S’not Funny
35 ALZHEIMER’S DISEASE: THE WAY FORWARD 105 Universal Flu Vaccine: Coming Soon
37 GENE THERAPY TARGETS AIDS To A Pharmacy Near You?
39 FROM DRUG DISCOVERY TO APPROVAL: PHASE IV 107 CRISPR Babies
41 FROM DRUG DISCOVERY TO APPROVAL: PHASE III 109 Biomanufacturing: How Biologics Are Made
43 FROM DRUG DISCOVERY TO APPROVAL: PHASE I/II 111 Wading Deeper Into Biomanufacturing
45 THE MANY FACES OF BOTOX 113 Stump The Clumps: New Drugs
47 KEEPING CANCER IN THE CROSSHAIRS To Treat Amyloidosis

49 NEW GENE THERAPY DRUG EXPLAINED 115 The Race Against NASH
51 VIRUSES TO THE RESCUE? 117 An Egg-Cellent Idea
53 COMING SOON? A NEW BIOLOGIC 119 An Epigenetic Liquid Biopsy
55 $8.7 Billion SMA Drug Explained 121 PARP1 Inhibitor Takes On Pancreatic Cancer
57 Papa’s Got a Brand New Lab Coat 123 The Molecular Causes of Obesity
59 Ticked Off: The Science Behind Lyme Disease 125 Cracking the Circadian Code
61 Immunotherapy Goes Viral 127 Breaking Down Drug Metabolism
63 Taking a Swing at Peanut Allergies 129 Swallowing A Biologic Drug?
65 CRISPR to the People 131 Taking A Swing At Allergies
67 CRISPR’s Genome Detectives 133 New Hope For Spinal Muscular Atrophy
68 Groundbreaking Migraine Drug Explained 135 GMO Apples Hit Marketplace

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137 Stem Cell Snapshot 218 From Drug Development To Approval: Phase IV
139 Therapeutic Antibody Primer 220 Market Access
141 The Mechanics Of Melanoma 222 From Drug Development to Approval: A Recap
143 The Intrigue Of HIF 224 Who’s Your Daddy? The Science of 23andMe
145 The PARP Race Is On 226 Meatless Meat: Biotech Burger Ain’t No Bean Patty
147 Pills, Proteins & Peptides 228 Alzheimer’s Disease: A Tough Nut To Crack
149 Putting The NA in DNA 230 Exploring Different Strategies to Fight Alzheimer’s
151 RNA Therapeutics March Onward 232 Cancer Diagnostic In A Drop
153 Decoding Your Genes 234 Turning On Cellular Garbage Disposals
156 Phage — More Than Just A Phase 236 Harnessing Your Immune System For Good
158 Viruses Blasting Cancer 238 Decoding The Gut-Brain Axis
160 Nanobodies: These Are Not Your Mother’s mAbs 240 Unwiring The Biology Of Fibrosis
162 Epigenome: Writing, Reading & Erasing 242 The Science Behind The Opioid Epidemic
164 A Skin Cell With Stem Cell Diversity? 244 The Molecular Cause Of Obesity
166 Cell Signaling Explained 246 Cancer Vaccines & Game Changers
169 Breast Cancer Subtypes 248 The Race To Beat SMA
171 Stopping A Big Problem: Blood Clots 250 Killing Cancer At Its Stem Cell Origin
173 Unpacking Digital Medicine 252 Post-translational Modifications In Biotech
175 Chaperoning The Rare Disease Dance 254 The First Three-Parent Baby
177 Putting The CAR-T Before The Horse 256 A Bone To Pick With Osteoporosis
179 The Science Of CRISPR/Cas9 258 DMD Makes The Cut
181 The Microbiome Magnified 260 The Tight Junction’s Function
183 Eye Of The Cytokine Storm 262 Zika On The Brain
185 The Multiple Myeloma Landscape 264 The Long And Short Of AMD
187 From Fantasy To Reality: Xenotransplantation 266 Vaccines: Schooling The Herd
189 Picturing Disease 268 The Next Generation Fight Against CF
191 Circadian Rhythm & Disease 270 The Antibiotic Economy
193 DNA Vaccines Explained 272 A Drop Of Gene Therapy
195 Natural Born Cancer Killers 274 Breaking Bad With NSCLC & SCLC
197 Off-Color: The Science Behind 276 The Plight & Promise of p53
Color Vision Deficiency 278 Biotech In Space!
199 The Science Behind Opioid Addiction 280 Without A Treg To Stand On
201 Bye-Bye Opioids? Introducing Electroceuticals 282 On A Tumor’s Turf
203 Zinc Finger Nucleases 284 Next-Generation CAR-T
204 Plants That Heal 286 Hacking The Immune Response
206 Drug Discovery 101 288 Pushing The Self-Destruct Button
208 Drug Discovery 201 289 Attack Of The Migraine
210 Gene Therapy Cures 291 Disrupting The Cycle
212 Drug Discovery 301 293 An Inborn Error Of Metabolism
214 From Drug Development to Approval: Phase I/II 295 Two Monoclonal Antibodies Walk Into The Market
216 From Drug Development To Approval: Phase III 297 Unmasking Multiple Sclerosis

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299 On The Hunt 373 Targeting Breast Cancer
301 Attacking All Angles Of Alzheimer’s 375 The Science Behind The Blockbuster
302 The Emerging Proteomics Market Drugs Of Tomorrow

304 Biotech Battles Zika 377 Getting Rid Of Allergies

305 Stomach Stapling Paves A Pathway 379 Three Parent Babies Approved
307 Finding Footprints 380 The Power Of Oncolytic Viruses
309 Drugging The Undruggable 382 Interchangability Denied To First
US Approved Biosimilar
311 Interrupting HIV
384 It’s A Great Time To Be In Biotech
313 Detecting A Silent Killer
386 New Brain Cells From A Pill
314 The Medicine Machine
388 GMO A Day Keeps Doc Away?
316 A Foodie’s First-In-Class FDA Approval
Thanks To RNAi Apples
318 Catching The First Drop Of Cancer
390 Curing Hepatitis C
320 The Uber Of The Human Body?
392 The State Of Cystic Fibrosis And Precision Medicine
321 The ABCs Of Hepatitis
394 Longevity In Biotech
323 Cancer Immunotherapy Goes Viral
396 Designer Genes: An Introduction
325 A Big Pain To Genome-Editing
327 Et Tu, Immune System? 398 Biosimilars: Ready or Not, Here They Come
329 Is Genetic Variety The Spice Of Life? 400 The Bispecific Antibody: A Lethal Hybrid
331 The Science Behind The Nobel Prize 401 Hitting The Mark With Alzheimer’s
333 Presenting The New Class Of Drug Development
Transcriptional Therapeutics
403 Capturing The Running Backs Of Cancer
334 Breaking Down Lung Cancer 405 12 Biotech Concepts Everyone Should Know
336 Feeding Cancer 406 Pioneers In The Land Of Small Proteins
338 The Powered Exoskeleton 408 Driving Down The Epigenetic Highway
340 Deciphering DMD 410 Breaking From The Pack With Cancer Diagnostics
342 RNAi Crashes The PCSK9 Party 411 Biotech’s Battlefront: Monoclonal Antibodies
344 Dodging Another Ebola Outbreak 413 Pursuing The Promise Of Unlimited Platelets
346 Stopping The Hedgehog 415 Sculpting A Better Life For Sickle Cell Patients
348 A Drop Of Biotech 417 Mutations And Disease: The Spice Of Life
350 The Realm Of Next Generation Sequencing 419 Squeezing The Juice Out Of Drug Metabolism
352 Targeting The Cause Of Cystic Fibrosis 421 Tailoring Stem Cells To Fashion
354 What The Heck Is qRT-PCR Diagnostics? Replacement Organs
356 Potential Of PCSK9 Inhibitors 423 Can Cell Death Keep Cancer Patients Alive?
358 The Danger Of Antibiotic Resistance 425 Matters Of The Heart
360 Immune System Checkpoint Therapies On The Case 427 Breaking The Ice With ALS
362 A Health Crisis In The Making 429 Cerdelga: 25 years In The Making
364 Antisense, RNAi And MicroRNA Explained 431 A Chip Off The New Block
366 Inching Across The Blood-Brain Barrier 433 Cultivating Biologics From Plants?
368 Packing A Punch With The Immune System 435 Tiny Virus, Big Problem
370 Ready. Set. CRISPR. 437 Playing Detective With Checkpoint Therapies
372 The Rise Of Nanomedicine 438 Sci Fi Science: Cool In Movies, Cooler In Clinic

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440 Dissolution Of Cancer Club Wanted 452 MRSA vs. Dalvance: Let’s Get Ready To Rumble
441 Inhaling Insulin Is A Breath Of Fresh Air 454 Don’t Be DOA. Learn The MOA.
443 Biotech Cake At Your Next Gluten- 456 Mitochondrial Medicine Going Mainstream?
Free Birthday Party? 458 Genome Editing: Curing HIV?
445 Blunt Approach: Fecal Transplants 459 RNAi Advantage Goes To Dicerna
447 Science Geeks Rock! 460 No Free Lunch For Obesity Drugs
449 Scabbing Now A “Walk In The Park” 461 200,000 Is The Magic Number
For Hemophilia A Sufferers
451 Saved By Screening

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HOPE ON THE HORIZON FOR HUNTINGTON’S Life Science Training from Industry Experts

HOPE ON THE HORIZON The CNS—the brain and spinal cord—works with the
FOR HUNTINGTON’S peripheral nervous system. This vast network feeds into
every tissue and funnels data to central command. The
No question about it, Huntington’s disease (HD)
signals received from nerve cells enable both voluntary
is cruel. In the beginning, sufferers of this fatal
and involuntary movement. They also allow the brain to
neurodegenerative disorder experience involuntary
process and interpret sensory information.
movement and mood disturbance, most often in the
form of depression. As the disease progresses, patients Specialized cells called neurons convert chemical
lose the ability to walk, speak, and eventually even messages into electrical signals that convey information
swallow. Life expectancy after symptoms first appear is throughout the nervous system. Dendrites, branch-like
about twenty years. Ninety percent of HD cases affect structures, take in chemical messages down through
adults between the ages of 30 and 50. The remaining ten the cell body (soma). The long, tail-like extension on the
percent of patients are even younger, and the course of other end of the neuron is an axon. They are insulated
their disease faster. by a fatty membrane known as a myelin sheath. This
protective coating also helps speed electrical signals
As if to recognize Huntington’s Disease Awareness
through the axon terminals.
Month (each May), there is some good news for
Huntington’s sufferers and their loved ones. Last week,
Roche (Basel, Switzerland) and Ionis (Carlsbad, CA)
scientists published a paper describing their promising
Phase I/IIa study of a new Huntington’s drug temporarily
labeled RG6042. The study demonstrated that the
new drug is safe. Researchers also observed some
preliminary signs of efficacy in the small number of
participants tested.
Larger, more extensive tests are required before
anyone can definitively say that the drug works. But
even this small study provides hope in the face of this
devastating disease. Neurotransmitters are data delivery guys. They are
This week, we review the basics of nervous system chemical messengers that work in the spaces in between
function and explain Huntington’s disease in more detail. neurons, the synapses. When the neuron’s dendritic
Then we’ll describe the Roche treatment and others branches encounter a neurotransmitter, ion channels in
in development. the neuronal cell membrane open. These minute gaps
allow positively charged sodium ions to enter a cell. The
THE BODY ELECTRIC positive charge initiates an electrical signal through the
neuronal body and down the axon tail, releasing other
Our nervous system is incredibly complex. It stretches
neurotransmitters into a neighboring synapse. This
into the body’s every nook and cranny, controlling and
process creates a cascade of neurotransmitters down
receiving all of the outside world’s signals from a central
the neuron chain, with different neurons sending and
command station. Like any other part of us, the central
receiving different neurotransmitters.
nervous system (CNS) falls prey to diseases we don’t
always understand.

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 RG6042 MECHANISM OF ACTION
RG6042 is an antisense drug, This approach aims to
reduce how much protein a specific gene produces—in
this case, the mutated huntingtin gene. RG6042 operates
by destroying the gene’s RNA.
Recall that the information in genes is first converted to
RNA, which is then translated to a protein. This approach
works on the premise that impeding the production
of mutated huntingtin will result in fewer of HD’s
heartbreaking symptoms.
Antisense therapeutics utilize existing cellular pathways
The billions of neurons within the CNS—communicating
that target and destroy double-stranded RNA (dsRNA).
via hundreds of different neurotransmitters—regulate
To activate the pathway, researchers introduce a piece
just about everything: our movement, hunger,
of modified RNA with a sequence that complements the
temperature, emotion, and arousal. Everything!
mutated huntingtin RNA. With RG6042, this synthetic
SLOW KILLER IN ACTION RNA is the drug. Because its sequence is complementary
to the mutated huntingtin RNA, this new drug binds it.
Huntington’s Disease is an attack on this electric power
The cellular enzyme RNAse H then cuts up the RG6042/
grid. It’s monogenic, caused by a mutation in one gene—
mutated huntingtin RNA hybrid. No mutated huntingtin
the huntingtin gene. The disease is also autosomal
RNA, no mutated protein.
dominant: if one parent has HD, their children have a
50 percent chance of getting it. Because the disease has
no cure, some at-risk individuals struggle over whether
to test for the gene. Unless symptoms of juvenile-
onset HD are present, testing for HD before age 18 is
prohibited. The intent of the restriction is to help ensure
those tested more fully understand the implications for
their future.

TERM OF WEEK:
TRINUCLEOTIDE REPEAT
The mutation that causes HD is a trinucleotide repeat.
Three nucleotides (CAG) are repeated in the middle of In the Phase I/IIa trial, patients exhibited reduced
huntingtin gene (HTT). Even normal versions of HTT have amounts of mutated huntingtin protein in their
the CAG trinucleotide repeat. However, fewer than 36 spinal fluid.
repeats means the disease is not present. Higher than 40
means the disease manifests; copy numbers between 36 OTHER DRUGS IN DEVELOPMENT
and 40 means the individual may or may not be affected.
Until recently, the function of HTT and how its mutations
Generally speaking, the higher the number of repeats
cause neuronal cell death were largely unknown.
beyond 40, the earlier disease onset occurs and the
Recent research from the University of California San
more quickly HD progresses.
Diego suggests that HTT plays a role in activating gene
expression in neurons through its interaction with the
protein PPARδ. The protein’s function is significantly
diminished in HD neurons, possibly because of
interaction with the mutated HTT protein.
Researchers identified an experimental drug, KD3010,
as boosting PPARδ activity. This product was tested in
a mouse model of HD and found to significantly reduce

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neurodegeneration. Even better news—the drug has conducting preclinical research on zinc-finger mediated
already passed human safety trials for diabetes. That will genome editing in HD.
likely speed KD3010 trials for HD. Huntington’s disease is a formidable opponent for both
Because HD is a single gene disorder, it makes a good its sufferers and the health care professionals struggling
candidate for gene therapy and genome-editing to help them. These latest technologies might just be
approaches. Sangamo Therapeutics (Richmond, CA) is powerful enough make a difference.

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THE INFINITESIMAL ENGINES OF BIOLOGICS Life Science Training from Industry Experts

Last week, we examined cell lines—what they are, where critical post-translational modifications—chemical and
they come from, and how companies use them in the physical changes made to a protein by cellular enzymes
discovery phase of drug development. This week, we after the protein is produced by the cell. Bacterial cells
turn to the critical role of cell lines in making those drugs. lack the enzymes required to make those changes.
Although all drugs restore or improve our health, Enter the humble Chinese hamster ovary (CHO)
the methods used to manufacture them vary greatly. cell. These cells were already commonly used to
Small molecule drugs are chemically synthesized; produce human proteins for drug discovery research.
the production of biologics revolves around cells. Consequently, they made an obvious choice for
Pharmaceutical scientists “program” cells to make biomanufacturing complicated proteins such as
proteins by transferring the genes that code for them antibodies and enzymes. The first biologic drug
into the cell line used for production. produced in CHO cells was Activase (Genentech), which
came on the market in 1987. Activase is an enzyme
ONCE UPON A BACTERIA… that dissolves blood clots. Today, CHO cells provide the
Way back in the days of disco and polyester, two platform for about 70% of biologic drugs.
California scientists discovered two phenomena that Why use CHO cells and not a human cell line?
ushered in the age of biotech. Stanley Cohen of Stanford Pharmaceutical companies actual prefer to employ
University (Palo Alto, CA) and Herbert Boyer of non-human mammalian cell. That’s because viruses
University of California, San Francisco found out that: that infect hamsters are unlikely to infect human
1) It’s possible to transfer genes for human proteins into cells. Yet as mammalian cells, CHO cells are capable of
bacterial cells and 2) the recipient bacteria can “read” the producing complex human proteins. These cells can also
gene and translate it into the protein. grow in suspension. Because they don’t need to attach
Cohen and Boyer realized that, essentially, human themselves to a solid surface such as the bottom of a
genes can serve as recipes for bacteria to make human tissue culture flask, it’s possible to grow thousands of
proteins. Venture Capitalist Robert Swanson convinced liters of CHO cells in enormous vats.
Boyer that the technology was commercially viable.
Together, they founded Genentech (South San Francisco, ON BEYOND HAMSTER CELLS
CA) in 1976. The company’s first product, human insulin, Like in any area of biopharma, innovation still occurs in
was manufactured in E. coli bacteria. The Food and Drug cell line development, despite the workhorse CHO cell.
Administration approved this first ever biologic drug in Here’s just one of the more recent cell lines that may
1982. Other human protein therapeutics produced in prove more efficient than CHO:
E. coli include human growth hormone, interferon, and Dyadic International ( Jupiter, FL) has bred the C1 cell
parathyroid hormone. line from the Myceliophthora thermophila fungus. These
cells have been used to produce industrial enzymes since
HAMSTERS AND OVARIES the 1990s. In fact, Dyadic started out producing enzymes
AND BACTERIA—OH MY! for the stonewashed jeans that have gone in and out of
Biologics production in E. coli proved extremely fashion for decades.
successful. However, to grow, the biotech industry
In recent years, the company turned its attention to
needed other cellular vessels. That’s because E. coli
perfecting the cell line for biopharmaceuticals. In the
successfully produce only those human proteins with a
words of CEO Mark Emalfarb, the company has gone
relatively simple structure.
“from jeans to genes.”
Drug scientists tried to make larger and more complex
C1 cells are now able to correctly produce a wide range
proteins with E. coli. However, they found that the
of human therapeutic proteins, including monoclonal
resulting proteins lacked the correct structure and

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antibodies, which have long been the exclusive domain project, to facilitate fast and effective responses to
of CHO cells. new infectious diseases. The idea is that using C1 cells
This cell line potentially offers significant savings in could expedite the production of proteins for drugs
time and money. The cells grow much more speedily or vaccines. Just last week, Dyadic announced that the
than CHO, doubling in two hours versus twenty hours. Israel Institute for Biological Research selected it to
Moreover, their growth media costs far less than that help develop treatments and vaccines for the current
used for CHO cells. Emalfarb estimates a savings of five coronavirus, COVID-19.
to 10 times that of what it takes to use CHO cells. Further, Whether bacterial or hamster or fungal, the humble
the amount of protein produced per liter of cells is cell provides the foundation for manufacturing today’s
much higher for C1 cells than for CHO cells, according to groundbreaking new therapies. Over the next several
Dyadic’s website. years, expect to see new and improved cell lines making
The company is working with the Zoonotic Anticipation biologic production even faster and more cost-effective.
and Preparedness Initiative (ZAPI), a European Union

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EX-CELL-ENT RESEARCH PARTNERS Life Science Training from Industry Experts

Cells are the basic structural and functional unit of all solid surface such as the bottom of the flask. In contrast,
organisms. Without them, drug development would be suspension cultures can grow in the entire volume of
stuck in the bad old days before antibiotics. Researchers medium. Once established, a cell line can be produced in
need squillions of these membrane-bound bundles of quantity or frozen for later use.
molecules to develop new medicines and to make sure
they’re safe. If you’ve worked in or around biopharma, THE BIRTH OF HUMAN CELL LINES
you’ve likely heard the term “cell line” before. Every Researchers have been establishing cell lines for about
phase of drug discovery and development hinges on cell a century. American Type Culture Collection (ATCC;
lines. This week and next, we’ll take a look at them: their Manassas, VA), a nonprofit organization which collects
origins and uses. and distributes cell lines, was established in 1925. Some
species, such as rodents, give rise to cell lines relatively
TERMS OF THE WEEK: CELL easily. With others, the process is much more trial-and-
CULTURE AND CELL LINE error. Dr. George Gey of Johns Hopkins University
Cell culture is the process by which cells are grown under (Baltimore, MD) produced the first human cell lines
controlled conditions outside their usual “habitat,”—be in 1951. An oncologist, Gey tried to accomplish this by
it rat, insect, or human body. To create a cell culture, isolating cells from his patients’ tumor biopsies. Most of
scientists start with a bit of tissue and break apart its these attempts failed—until the cervical cancer biopsy
cells with enzymes. The cells go into a flat plastic flask of Henrietta Lacks. Her cells proved to be tremendously
of growth medium—a concoction of nutrients such amenable to growing in the lab, and established one
as amino acids, carbohydrates, salts, vitamins, and of the most widely used cell lines within biomedical
minerals. Depending on the type of cell, the growth research today: HeLa cells.
medium may also contain growth factors and hormones.
The flask then goes into an incubator set close to body IMMORTAL CELLS
temperature—37 degrees Celsius. Because HeLa cells came from a cancer biopsy, they’re
“immortal,”—capable of living endlessly in the lab. Cell
lines from healthy tissues can divide only a limited
number of times in culture before they die.
To persist indefinitely, cell lines must be “transformed”
into immortal cells. Scientists infect the cells with
certain viruses, which disrupt genes that regulate
their growth. Cells from tumors and those enhanced
with viruses possess characteristics that differentiate
them from healthy cells. Researchers who want to
study normal cells use a different method to prolong
their viability. They add a gene that codes for “human
telomerase reverse transcriptase.” This gene provides
Scientists call the cells that develop from the initial
cells with an enzyme that lengthens telomeres. These
isolation and growth phase primary culture cells. After
short sequences of DNA appear at the end of all
they’ve multiplied to cover the entire bottom of the flask,
chromosomes. Extending their length extends the life of
they’re ready for transfer to a bigger container. At this
the cell.
point, they’re referred to as a cell line. Most cell lines are
“adherent,” which means they attach themselves to a

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THE MINUTE CONFINES OF DRUG SUPER CELL LINES: IPSCS
DISCOVERY RESEARCH Induced pluripotent stem cells (iPSCs) are the rock
The biotech applications of cell lines are seemingly stars of cell culture. “Pluripotent” means that these
infinite. For example, oncologists examine cancer cell cells possess the amazing potential to become any
lines to understand the mechanisms of a particular adult human cell type. They begin as humble and
cancer or screen potential drug candidates. Non- highly accessible adult human skin cells. Activating
cancerous cell lines may be used in drug safety testing. certain genes in the lab turns these humdrum cells into
To study a virus in the lab, scientists must find an biopharmaceutical magic. iPSCs have become invaluable
“infectible” cell line to see how it interacts with its host. to some research efforts. In Alzheimer’s disease, for
For example, scientists studying a respiratory virus example, it’s impossible to get biopsies of patients’
may use A549 cells, a common lung cell line. Finally, brains. Now, researchers can convert skin cells from an
researchers use cells to make proteins for study, using Alzheimer’s patient into an iPSC and then coax them
recombinant DNA technology to transfer the gene into into becoming brain cells. Because they have the same
the cell. Chinese hamster ovary (CHO) cells and human genetics as Alzheimer’s patients, scientists can use them
embryonic kidney cells (HEK293) provide particularly rich to model the disease in the lab.
grounds for producing human proteins. Next week, we’ll turn our focus to cell lines used in
biomanufacturing. We’ll explore the tried and true CHO
cells and take a look at a new cell line that may offer
some new advantages.

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SEARCH AND DESTROY: CHIMERIC
ANTIGEN RECEPTOR T-CELLS Life Science Training from Industry Experts

The first (and so far, only) chimeric antigen receptor it to evade detection. This trickery is known as
T-cell (CAR-T) therapies—Kymriah (Novartis; Basel, the tumor microenvironment and once again the
Switzerland) and Yescarta—(Gilead; Foster City, CA) dangerous cancer cells can pass by undetected.
were approved in 2017, and remain one of the hottest So what’s a scientist to do?! Figure out a way to train
immunotherapies on the market today. They work by killer T-cells to ALWAYS recognize and destroy cancer
boosting the body’s ability to recognize and attack cells…enter CAR-T.
cancer cells. This WEEKLY reviews the basics of CAR-T
technology. Next week, we’ll take a peek at CAR-T HOW TO TRAIN AN IMMUNE SYSTEM
innovations moving through the pipeline.
CAR-T therapies boost the body’s ability to recognize and
attack cancer cells. These “super” killer T-cells have been
TERM OF THE WEEK: KILLER T-CELLS
physically enhanced to go after cancer. Like the mythical
CAR-T therapy is based on a type of white blood cell chimera, this drug is composed of different parts.
called a killer T-cell. The job of these cells is exactly what Genetic engineers fuse an antibody with a killer T-cell
the name implies—to kill dangerous cells. They target receptor to create a chimeric molecule—the “C” in CAR-T.
diseased cells in the body via cell-surface receptors:
The transformation begins with technicians removing
each has a uniquely shaped receptor, and recognizes
killer T-cells from a patient’s body and isolating them in
its intended target because the shape of its receptor
the lab. Next, scientists use a viral vector—a virus that
“matches” or fits into a uniquely-shaped surface
has been modified to contain a therapeutic gene—to
protein found only on diseased cells. Once the killer
deliver a gene that encodes the chimeric receptor to the
T-cell “docks” onto its target, it injects an enzyme which
T-cells.
triggers death. The result: no more bad cells.
The enhanced receptor includes two parts: a targeting
domain and an activation domain. The first is the portion
that remains on the surface of the T-cell. It’s an antibody
that detects and locks onto a specific surface protein on
the patient’s malignant cells. The activation domain part
of the receptor is triggered once the targeting domain
attaches itself to the desired cancer protein.

WHY CAR-T?
In theory, our immune system should recognize the
unique proteins presented on cancerous cells. However,
there are two main reasons this doesn’t always happen:
1. Early on in the tumor development, the cell
composition is similar enough to healthy tissue that
it can be overlooked by the immune system. The engineered T-cells are then reinfused into
the patient.
2. Later, as a tumor progresses, it releases chemical
signals that suppress the immune response, helping Once back inside the patient, the targeting domain
finds the proper surface protein on the tumor cell and

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attaches to it. Then, the activation domain signals the transmits signals to the T-cell when it docks onto
killer T-cell to: its target.
• Stay alive. • T-Cell: A white blood cell that kills harmful cells.
• Make copies of itself (replicate).
• Release cytokines—chemical signals that activate TRICKY TERMINOLOGY
other white blood cells to assault the tumor. The medical community classifies CAR-Ts as a “cell-based
• Kill the target cell. gene therapy.” They’re immune cells that have been
engineered using gene therapy techniques.

IN THE PIPELINE
Two CAR-T therapies have currently been approved,
several more are in clinical development. The table
below shows a selection of CAR-T therapies moving
through the clinic:

WHAT’S INA NAME?

Chimeric antigen receptor therapy broken down:
• Chimeric: Composed of components from two
distinct parts, such as an antibody and a killer
T-cell receptor.
• Antigen: A protein that is recognized by an antibody,
such as a protein on the surface of a tumor cell. Biopharma never rests. Industry scientists are busy
creating the next generation of CAR-T therapies, which
• Receptor: A protein that is embedded in a cell
should prove to be both safer and more effective than
membrane and transmits signals to itself in response
the first generation. Tune in next week as we discuss
to being activated, for example a T-cell receptor
these new therapies.

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BLUEPRINT OF THE CENTURY: DNA Life Science Training from Industry Experts

Please enjoy this excerpt from our recently revised which he suspected were responsible for passing traits
book, The Biotech Primer: An Insider’s Guide to the Science from generation to generation.
Driving the Biopharma Industry. This book explains the Almost a century later, in 1944, scientists identified DNA
science behind the industry in an accessible manner, as the “particles of inheritance.” The race was then on to
providing the foundation needed to understand today’s determine its structure. Eventually, the research team
advanced therapies. of James Watson and Francis Crick solved this scientific
puzzle in 1953. The structure that Crick and Watson
DECODING THE MASTER PLAN conceived was a double helix. An X-ray image captured
At the dawn of the 21st century, the Human Genome by English chemist Rosalind Franklin revealed that
Project had just been completed. This revolutionary DNA was helical. Sadly, Franklin died before receiving
undertaking—determining the exact size, sequence, full credit for her critical contribution. Scientists had
and location of genes within the human genome, or proposed many other possible configurations, including
the full complement of DNA in each of our cells—is a triple helix. However, only the double helix model fit
one of biology’s greatest achievements. For the first the evidence of base pairing provided by Erwin Chargoff.
time, researchers possessed the complete human
blueprint. Ideally, they could now pinpoint the
source of countless diseases. This ushered in what
many, including the European Commission and the
National Academy of Sciences, have referred to as the
Century of Biology. Since then, the ability to quickly
and economically sequence the human genome has
increased exponentially.
DNA is the blueprint of this Century of Biology. Found
in all living cells, DNA is the genetic material that stores
and transfers information. In this chapter, we look deep
inside the cell and examine the molecule upon which the
entire biotechnology industry rests, starting with key
discoveries and the scientists behind them.

THE STORY OF DNA

In the 1850s, an Austrian monk named Gregor Mendel
performed breeding experiments with pea plants.
He observed that certain genetic characteristics
were passed down from one generation to the next
in specific ratios. Because he was the first to analyze The last sentence of Watson and Crick’s Nature paper
the inheritance of traits systematically, many consider that described the double helix structure, made a critical
Mendel the father of genetics. Mendel’s contributions point: “It has not escaped our notice that the specific
are even more impressive because though he didn’t pairing we have postulated immediately suggests a
know about DNA, he predicted its existence. He called possible copying mechanism for the genetic material.”
what we know now as DNA “particles of inheritance” This suggestion would have profound implications for
the as-yet-unrealized field of biotechnology.

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LIFE’S BUILDING BLOCKS
The full name for DNA is deoxyribonucleic acid. It’s a
long molecule made up of repeating subunits called
nucleotides. Each nucleotide consists of three parts:
1. A sugar molecule called deoxyribose the “D” part of
DNA. This molecule has five carbon atoms that form
a ring structure.
2. A phosphate molecule attached to the fifth carbon
atom in the ring. This gives DNA weak acidic
properties and is the “A” in DNA.
3. A molecule called a base attached to the first carbon
atom in the ring. There are four bases: adenine (A),
cytosine (C), guanine (G), and thymine (T).

The prefix “deoxy” means “without oxygen.” It refers to

the lack of an oxygen molecule at a particular spot in the
sugar molecule. “Ribo” stands for ribose, which describes
the particular sugar. All organisms use the same
bases—A, C, G and T, and the same sugar and phosphate
molecules. In other words, the nucleotide building blocks
for the genetic material of all forms of life are identical.

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PUTTING IT ALL TOGETHER
Nucleotides are linked to form a long chain. During DNA
synthesis, a cellular enzyme called DNA polymerase
links the phosphate group of one nucleotide to the sugar
group of another. This forms what is often referred to as
the sugar-phosphate backbone.

These four tiny bases—A, C, G, and T—come together to

form the blueprint of life. Understanding this blueprint
has enabled all of the incredible advances we’re now
experiencing within biopharma, including full genome
sequencing, gene therapy, and genome editing.

COCKTAIL FODDER
But how do nucleotide chains form a double helix Where do cells get the nucleotide building blocks to
structure? An early experiment by Chargoff revealed make DNA? The food we eat, such as chicken, fish, fruits,
some striking patterns. He noticed that within a DNA and vegetables, are made of cells. Animal and plant cells
molecule, the number of Cs is always the same as the contain DNA made of the same constituent parts as
number of Gs. Similarly, the number of Ts is always ours. The DNA you consume in your salad or salmon is
exactly the same as the number of As. This led to the broken down into the nucleotides we need. Our cells can
conclusion that DNA is composed of paired strands: Cs also synthesize the four nucleotides de novo, meaning
on one strand are matched to Gs on the other, and As on “from the beginning,” using atoms from proteins, sugars,
one strand are matched to Ts on the other. The pairing and fats that we consume.
of Cs to Gs and As to Ts between strands is accomplished
by chemical bonds. Their geometry, determined by the
particular shapes of the nucleotides themselves, gives
DNA its iconic shape.

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SPOTLIGHT ON MACULAR DEGENERATION Life Science Training from Industry Experts

Getting old is for the birds. Time has its way with nearly Ophthalmologists and other scientists don’t know
every part of us: skin, hair, muscles. Few changes are exactly what causes AMD. It is however, associated
more alarming, however, than those to our eyes. There’s with a buildup of proteins and lipids just beneath the
more to old eyes than bifocals and cataracts though. retina. These deposits, drusen, are a normal part of
One of the most common eye diseases is age-related aging. However, the presence of larger or more drusen
macular degeneration (AMD). It affects more than 13 raises the risk of AMD. As the disease progresses, vision
million people over age 50 in the United States. It’s decreases. There are two types of AMD: neovascular
the leading cause of vision loss in this age group. This (wet) or atrophic (dry).
WEEKLY takes a closer look at AMD and the ways in
which the biopharma industry is working on treatments. WET AMD
Eyes are spectacularly complicated organs: they contain In wet AMD, the infiltration of excess blood vessels is the
over two million moving parts. You know some of the main culprit. These abnormal vessels often leak fluid and
basics: iris, pupil, lens, retina. In some ways, this last, the blood, injuring the retina. Wet AMD progresses quickly,
retina, is more accurately considered part of the brain. leading to loss of central vision without treatment.
This form of the disease accounts for about 10 percent
TERM OF THE WEEK: RETINA of cases.
The retina is composed of light-sensitive nervous tissue FDA-approved treatments for wet AMD include
which forms a thin membrane that lines the rear two- Lucentis (Genentech; South San Francisco), Beovu
thirds of the eyeball. It takes in light from the world (Novartis; Basel, Switzerland), and Eylea (Regeneron
around and converts into neural signals that travel along Pharmaceuticals, Tarrytown, NY).
the optic nerve to the brain, telling us—“Hey, there’s an These work by mopping up excess vascular endothelial
apple, or a laptop, or whatever.” growth factor (VEGF), which causes the excess blood
vessel growth. Lucentis and Beovu are monoclonal
antibodies specific for VEGF. Eylea consists of the VEGF-
receptor fused to the constant region of an antibody
for stability. Similar to a mAb, this combination is called
a fusion protein. It’s highly specific for VEGF, binding it
before it reaches its intended receptor on the surface
of blood vessels. These VEGF-blocking treatments
effectively stop the progression of AMD, but don’t cure it.
Kodiak Sciences (Palo Alto, CA) is working on an
antibody-biopolymer conjugate (ABC) called KSI-301,
which also targets VEGF. An ABC is an antibody with a
biopolymer—a chain of repeating subunits produced by
The macula is the small central area of the retina that
a living organism—attached. In KS-301, the repeating
enables central, high-resolution, color vision.
subunits are lipids. They make the antibody more stable.
That means patients can go up to five months between
A GRADUAL LOSS OF VISION
injections, compared to one month (Lucentis), two
Macular degeneration progressively devastates eyesight, months (Eylea), or two to three months (Beovu). The
causing blurred vision and blocks the center of a treatment is now in Phase II/III development.
person’s visual field. What begins as a minor annoyance
These therapies are injected in the patient’s eye,
ends up making everyday tasks such as reading and
which makes them understandably unappealing. As
driving impossible.
an alternative, PanOptica (Mount Arlington, NJ) is

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developing PAN-90806. Patients could administer this binds to complement protein C3 and prevents
small molecule inhibitor of VEGF at home by eyedrop. its interaction with other complement proteins.
The drug has completed Phase I clinical studies. Preventing this interaction impedes activating their
RegenxBio (Rockville, MD) aims to do away with destructive power.
repeated treatments altogether with gene therapy. RGX- • IVERIC Bio’s (Princeton, NJ) DNA aptamer Zimura is
314 delivers a gene encoding an anti-VEGF antibody. In in Phase II development. DNA aptamers are short
Phase I/II studies, participants produced the therapeutic strands of DNA that bind to a specific protein. Zimura
protein, which controlled VEGF levels. binds to and inhibits complement protein C5.
• Ionis’ (Carlsbad, CA) and Roche’s IONIS-FB-LRx is
DRY AMD an antisense drug that blocks the production of
Dry AMD involves a gradual breakdown in the macula’s complement factor B, and is currently in Phase II
light-sensitive cells. The dry variety progresses much clinical testing.
more slowly than wet and accounts for about 90 percent
Last month, the National Eye Institute announced
of AMD cases. Advanced dry AMD occurs when cells
preparations to begin clinical testing of stem cells for
in regions of the retina have wasted away and died.
dry AMD. In the lab, researchers have coaxed stem cells
Sometimes these regions of atrophy (death) look like a
to grow into retinal cells. In rodent and pig models,
map to the physician who is examining the retina, giving
these cells restored vision, setting the stage for testing
rise to the term “geographic atrophy” for late-stage
in humans.
dry AMD.
Currently, no treatments exist for dry AMD. However, JUVENILE MACULAR DEGENERATION
some studies suggest that high doses of antioxidants Most macular degeneration is connected to aging.
including C and E vitamins, copper, zinc, and beta- However, Stargadt disease and other juvenile forms are
carotene may slow its progression. associated with several different genes. Also referred to
Inflammation, specifically the activation of complement as macular dystrophy, all juvenile varieties are inherited.
proteins, is associated with drusen buildup and dry AMD They have no cure or treatment. The therapies under
progression. When they’re activated, these immune development for dry AMD may also prove useful in
system proteins interact, destroying targeted cells. Stargadt disease.
Inappropriate activation of complement proteins can Biopharma is hard at work developing more
result in the destruction of healthy cells and tissue. patient-friendly treatments for wet AMD and the
A number of drugs in clinical development work by first ever treatments for dry AMD. Look for future
inactivating complement proteins: developments here.
• Apellis Pharmaceuticals’ (Crestwood, KY) peptide
drug APL-2 is in Phase III development. APL2

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GETTING THE JUMP ON ALZHEIMER’S DISEASE Life Science Training from Industry Experts

GETTING THE JUMP ON provide useful information. Unfortunately, the only way
ALZHEIMER’S DISEASE to truly diagnose AD is by autopsy.

Your dad has started losing his car keys, forgetting The need to develop accurate, minimally-invasive
important dates, and even more alarming —is getting diagnostics is clear. Better tests will, in turn, support
lost on the way to the grocery store. You’re thinking, improved clinical testing of potential drugs, as identifying
“Is it Alzheimer’s?” Maybe it’s time for a trip to the patients becomes easier. Recognizing the need for
doctor’s office. better AD diagnostics, the Alzheimer’s Drug Discovery
Foundation(New York, NY) has launched a Diagnostics
Unfortunately, that trip may prove frustrating
Accelerator. With backing from the Gates Foundation
because diagnosing Alzheimer’s disease (AD) isn’t
(Seattle, WA) and other philanthropic donors, they so far
straightforward. First, cognitive decline in general, and
have committed $30 million to funding research at the
dementia more specifically, have many possible causes.
cutting edge of AD diagnostics development. Here’s an
Second, there’s no one test for AD.
overview of some of the research they are funding.
That’s increasingly a problem. About five million people
in the US now suffer from AD. By 2025, the number BLOOD AND PROTEIN
of people over age 65 with AD may reach more than AD has long been associated with the abnormal buildup
seven million (Alzheimer’s Association). This condition of two neurological proteins: amyloid-beta plaques
poses a profound public health threat and a formidable (Aβ) and tau. Damaged neurons release a third, called
challenge to biopharma companies. neurofilament. It may also be an early indicator of AD
Previous issues of the WEEKLY have looked at AD’s if it appears with the others. A number of research
underlying pathologies and explored the latest groups are developing blood tests for these proteins.
pharmaceutical interventions. This week, we turn to They’re validating the tests by comparing samples from
the other half of the battle against this heartbreaking cognitively normal and AD patients of the same age and
condition: creating accurate, minimally-invasive by correlating them with brain scans.
diagnostics. Who knows, maybe one day a simple blood • The University of Gothenburg (Sweden) is a
test or retinal scan will reveal the presence of AD soon hotbed of research into next-generation protein-
enough for early intervention. based AD diagnostics. Drs. Kaj Blennow and Henrik
Zetterberg’s work led to the use of spinal taps to
HOW DIAGNOSIS WORKS detect AD-associated proteins. Now, teams in their
TODAY (OR DOESN’T) labs are developing and validating similar tests from
Back to the doctor’s office. If the doctor suspects AD, blood samples. Dr. Zetterberg’s lab is partnering with
she may assess your dad’s cognitive function or check Roche Diagnostics (Rotkreuz, Switzerland) on a test
his reflexes. If he performs poorly, the doctor needs to to detect Aβ40 and Aβ42 protein fragments. When
rule out other possible causes, such as a vitamin B-12 present in high amounts, these proteins cluster to
deficiency, depression, or stroke. She may order brain form amyloid plaques. The Gothenburg researchers
imaging to check for abnormalities such as high levels of are also looking into spotting these proteins in saliva.
amyloid-beta protein, a hallmark of AD. However, brain
• Eisai (Tokyo, Japan) is also developing blood tests
imaging is pricey. Further, most insurance plans won’t
for Aβ40 and Aβ42. In particular, its researchers are
reimburse them because current AD treatments don’t
looking at the ratio of the two proteins. They suspect
differ from those for other types of dementia. Patients
that reduced levels of Aβ42 may indicate early AD.
sometimes qualify for a spinal tap to detect tau or
amyloid-beta proteins in their blood, which can indicate • Randy Bateman at Washington University (St. Louis,
AD. But no one wants a spinal tap. These tests may MO) is also leading studies that examine ratios
between these two proteins as an early marker

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 of AD. In one study, some patients were initially (Monmouth Junction, NJ). These detect differences in
classified as false positives based on a comparison specific microRNA levels in people with AD.
of blood test and brain imaging results. The blood
test suggested they had AD; the brain image said RETINAL SCANS
they didn’t. Repeat brain imaging studies four years The retina connects directly to the brain via the optic
later showed signs of Aβ plaques, suggesting the nerve. Thus, it potentially serves as a sort of window
blood test may detect AD in the very early stages— into the brain. At Australia’s Center for Eye Research,
potentially a big boon to treating the disease. (CERA; Melbourne) researchers are zooming in on an
• University of California, San Diego neuroscientist eye scan to diagnose AD early. Developed in Dr. Peter
Doug Galasko is collaborating with ADX van Wijngaarden’s lab, the test detects amyloid-beta
Neurosciences (Ghent, Belgium) on a blood test deposits in the retina of healthy adults. It’s being
to identify low concentrations of the tau and developed for people with a family history of AD. One
neurofilament proteins. day it may replace costly brain imaging studies. Dr. Tom
MacGillivray at the University of Edinburgh and medical
BLOOD AND RNA imaging company RetiSpec (Toronto, CA) are carrying
Washington University researcher Dr. Laura Ibanez has out similar research.
identified 25 different types of protein-coding mRNA The range of basic science researchers and biopharma
associated with an increased risk of AD. She’s now companies working on minimally-invasive, less expensive
working to see if a blood test that analyzes these RNA tests for AD provides hope that one day soon, people at
molecules can predict the disease’s onset. risk for or in the early stages of AD can more easily be
Another kind of RNA—microRNA—forms the basis identified. This in turn should help with more efficient
of two blood tests now in development by Amoneta drug discovery for AD and ultimately a way to treat
Diagnostics (Huningue, France) and DiamiR Biosciences patients before the disease progresses.

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EXOSOMES: TINY VESICLES WITH BIG POTENTIAL Life Science Training from Industry Experts

EXOSOMES: BIO-TRASH electroporation—jolting the exosome with pulses of

TO BIO-TREASURE electricity—creates a temporary permeability that allows
the drug to enter the exosome.
We’ve all heard the phrase “One man’s trash is another
man’s treasure.” It turns out that what often holds true Exosomes loaded up with medicine would then be
for the antique business sometimes holds true for injected into the patient. The exosomes find their target
biotech as well. This WEEKLY shows how. tissue based on proteins present on that tissue’s surface.
The exosome’s lipid membrane is so similar to the cell’s
TERM OF THE WEEK: EXOSOME own membrane that it’s able to slip inside when they
come into contact, delivering its content to the cell.
There are these teeny tiny things each of our cells
secrete called exosomes. (Stay with us.) The term Exosomes display affinities for tissue, based on the
exosome derives from the Greek words for “external” location in the body from which they come. Researchers
and “body.” They’re only about one one-hundredth the control targeting by harvesting exosomes from cells
size of a typical human cell. They’re microscopic parcels known to produce exosomes that deliver their goods
secreted from cells and filled with cellular leftovers— to a specific tissue type. For example, exosomes from
RNA, proteins, and other remnants of metabolic pancreas cellscome equipped with “radar” that sends
processes. Microbiologists first discovered them in the them to lung cells.
1980s. Initially, the scientists thought exosomes were Codiak Biosciences (Cambridge, MA) is developing
a kind of cellular trash bag—providing a way to get rid another approach to getting drugs where they need
of the molecules a cell no longer needed. In the past to go via exosome. Their proprietary platform enables
decade, it’s become increasingly clear that exosomes them to engineer cells that produce both exosomes that
and what they carry play a role in intercellular signaling have specific contents and specific surface molecules
by delivering their contents to recipient cells. These that guide them.
tiny vesicles shuttle proteins and genetic information
Exosomal delivery makes an especially attractive option
between cells, acting as molecular mail carriers.
for drug makers with products that face significant
Exosomes play a role in activating the immune system,
delivery obstacles. These include RNA-based drugs that
and are likely involved in other aspects of health and
have struggled to achieve efficient delivery, and biologic
disease. Biopharma has turned its attention to figuring
drugs too large to enter cells. Aruna Bio (Athens, GA)
out how to use these miniscule containers to provide
has developed exosomes that ferry drugs across the
the means to deliver lifesaving medicines and aid in the
blood-brain barrier—another common stumbling block
diagnosis of disease. They may eventually even act as
to drug delivery.
stand-alone therapeutics.
FROM DELIVERY TO DRUG ITSELF
CATCHING A RIDE
The therapeutic potential of exosomes goes beyond
The most obvious application of exosomes builds on
drug delivery. Regenerative medicine specialists at
their role as cellular couriers to deliver drugs to target
Temple University in Philadelphia demonstrated
tissues that are not easily reached by conventional
that injecting exosomes from embryonic stem cells
methods, such as the brain, or to deliver drugs that do
can repair damaged cardiac muscle in a mouse model
not normally enter cells, such as monoclonal antibodies
of heart attack. Research at the University of New
or other protein therapeutics.
Mexico suggests that healthy cells near tumors produce
But how does a drug get inside these tiny vesicles to exosomes that kill cancer cells but leave normal
begin with? First, exosomes come from cells grown cells unharmed.
in the lab. Once harvested, technicians load them
with medicine in a number of ways. For instance,

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Biotech companies are following up on this basic science IN LIQUID BIOPSY
by developing exosome-based therapeutics. Codiak
Exosomes may also enhance the diagnostic power of
Biosciences plans to begin Phase I clinical oncology
of liquid biopsies. The idea is to capture exosomes
studies of its treatments this year. Evox Therapeutics
based on tumor-specific surface markers or to collect
(Oxford, U.K.) is preparing to enter Phase I studies
exosomes from blood or urine samples and identify
with an exosome-based therapy for lysosomal storage
them as cancer-associated by examining the genetic
disorders. Research at Aruna Bio focuses on exosomes
material inside. Aptly named Exosome Diagnostics
from neuronal stem cells that appears to reduce
(Waltham, MA) has an exosome-based urine test for
inflammation in the brains of mice and pigs and enhance
prostate cancer on the market. Biopharmaceutical
their function. The company plans to develop these
companies are still in the early stages of understanding
exosomes as therapies for stroke and amyotrophic
the power of the onetime cellular trash bag, but all the
lateral sclerosis (ALS). Finally, Celltex(Houston, TX)
signs point to a nanoscale revolution led by exosomes.
is also exploring the anti-inflammatory potential of
neuronal stem cell-derived exosomes as a potential
treatment for Alzheimer’s disease.

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NEW MUSCLE IN THE FIGHT AGAINST DMD Life Science Training from Industry Experts

NEW MUSCLE IN THE WHEN IT’S BEST TO BE A GIRL

FIGHT AGAINST DMD DMD occurs almost exclusively in boys. This is because
Last week’s biotech headlines lit up with the FDA’s the gene for dystrophin sits on the X-chromosome.
unexpected approval of Sarepta Therapeutics’ Females are born with two X chromosomes; males with
(Cambridge, MA) newest Duchenne muscular dystrophy one X and one Y. Therefore, baby girls end up with two
(DMD) drug, Vyondys 53. The FDA changed course copies of the dystrophin gene. As long as one produces
after rejecting the product in August because of safety working dystrophin, a child will have healthy muscles.
concerns, which the company has now addressed. Down the road, though, a girl with one faulty dystrophin
DMD is a genetic disorder that causes progressive gene may “gift” a son with a copy of it. This form of
muscle degeneration and weakness. It arises from a inheritance is called X-linked.
mutation in the gene that codes for a critical muscular About two-thirds of DMD cases result from inherited
protein. This ultimately leads to serious medical mutations; the rest arise from spontaneous
problems, including heart and lung problems. Typical genetic changes.
life expectancy for a child with DMD is 25 years. It
afflicts one in approximately 3,500 newborn baby TERM OF THE WEEK: EXON
boys worldwide. Vyondys 53 is an exon-skipping therapy. Exons are one
Today’s WEEKLY examines the science behind DMD and part of a gene’s structure. Recall that genes provide
explains the mechanism of Sarepta’s drug. the recipe to a cell for making a protein. When cellular
enzymes first read a gene, they produce pre-messenger
MUSCLE GLUE RNA (pre mRNA). This RNA contains segments that
DMD patients produce no functional dystrophin. aren’tinvolved in making the final protein. These
This protein forms part of a complex of proteins that segments are introns. The remaining bits of RNA are the
strengthen muscle fibers and protect them from injury exons. Once spliced together, exons form the final mRNA
during contraction and relaxation. Dystrophin connects that the cell converts to protein.
the proteins that make up muscle cells’ structural
framework with the network of proteins that surrounds
each cell. Physiologists and others sometimes refer to
dystrophin as the glue that holds muscle cells together.
Dystrophin also likely plays a role in transmitting
chemical signals within muscle cells.
Faulty dystrophin leads to damaged and progressively
weaker muscles. Almost from the start, kids with DMD
exhibit developmental delays. Over time, standing and
walking become impossible. Patients in their late teens
and twenties often develop cardiomyopathy or fatal
respiratory problems because the muscles that support
breathing deteriorate so profoundly.

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VYONDYS 53: HOW IT WORKS EASILY CONFUSED: SURROGATE
Vyondys 53 is an antisense therapy. This approach ENDPOINTS VERSUS
consists of administering short, synthetic RNA analogues CLINICAL ENDPOINTS
with sequences that complement the target RNA. In In clinical trials, an endpoint consists of an event or
this case, the target is exon 53 of the dystrophin pre- outcome that researchers can measure objectively to
mRNA. When Vyondys 53 enters cells, it binds to exon determine whether an intervention helps. There are two
53. This combination causes the enzymes that convert types: clinical endpoints and surrogate endpoints.
pre-mRNA into mRNA to skip over exon 53. The resulting A clinical endpoint indicates that a drug provides a
mRNA then produces a truncated version of the normal direct clinical benefit to the patient. In DMD, an example
dystrophin protein. In 2016, Sarepta won approval of a clinical endpoint would be a confirmed increase in
for Exondys 51, another exon-skipping treatment that muscle activity as a result of the treatment.
targets Exon 51 to treat DMD.
In some cases, companies can use a surrogate
endpoint to assess a treatment’s value instead of a
clinical endpoint. For Exondys 51 and Vyondys 53,
Sarepta relied on the surrogate endpoint of increased
dystrophin production. Surrogate endpoints allow the
FDA to approve new drugs for serious or life-threatening
diseases faster. If a drug is approved via a surrogate
endpoint, the FDA requires the manufacturer to conduct
follow-up studies to confirm that the product results in
a measurable clinical benefit. Sarepta is now running
these studies for both Exondys 51 and Vyondys 53.
The Biotech Primer WEEKLY will be taking a break for the
next two weeks in honor of the season. Happy Holidays
to all of our readers!

Both drugs target specific mutations so they won’t help

everyone with DMD. Roughly eight percent of DMD
patients have a mutation in exon 53 and should respond
to Vyondys 53. About 13 percent are likely to respond to
Exondys 51.

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CONTROLLING A KILLER, ONE STEP AT A TIME Life Science Training from Industry Experts

CONTROLLING A KILLER, • Another way to prevent infection is to block the

ONE STEP AT A TIME HIV surface protein, GP120, from binding the
CCR5 receptor. GlaxoSmithKline (London, UK) is
Last week, we reviewed the lifecycle of the human
conducting Phase III clinical testing of Fostemsavir, a
immunodeficiency virus (HIV). Learning how the virus
small molecule which targets GP120, the portion of
grows and develops has been crucial to developing
the viral surface protein that enables HIV to bind to
medicines that control HIV infection. This knowledge
and enter T-cells. Its developers believe that the bit
has enabled drug discovery researchers to devise ways
of GP120 that Fostemsavir targets mutates slowly,
to interfere with the virus at critical points in its life,
perhaps reducing the chances that resistant strains
preventing it from infecting more of the patient’s white
of the virus will emerge. Peptide therapeutics also
blood cells.
work on GP120. Roche’s Fuzeon interferes with
The resulting drugs—often referred to as highly active the ability of a portion of GP120 to fuse with the
antiretroviral therapies (HAART)—manage AIDS well. T-cell membrane.
Unfortunately, they don’t completely eliminate the
• Blocking viral replication: As described last
virus that causes it. In other words, they’re not a cure.
week, two steps are critical for viral replication:
In addition, HIV’s high rate of mutation means it rapidly
conversion of the viral RNA genome into DNA by
evolves resistance to drugs that once kept symptoms
the viral enzyme reverse transcriptase(RT) and
at bay. For this reason, HAART typically combines
insertion of this DNA into the host cell genome by
different drugs which is the “cocktail” approach. This
the viral enzyme integrase. The first antiviral drug
strategy takes advantage of the fact that viruses are
approved by the FDA for the treatment of HIV in
unlikely to develop resistance to multiple drugs at once.
1987, AZT, inhibited RT. Since then, many others
Researchers continue to work on new drugs that can
have been approved, including Viread and Emtriva,
successfully fight infection when resistance emerges.
both marketed by Gilead Sciences (Foster City,
This WEEKLY looks at how some current HIV medications CA). Click here for a complete list of RT inhibitors.
interrupt key stages of the viral lifecycle. Integrase inhibitors include Isentress (Merck & Co.),
Vitekta (Gilead Sciences), Tivicay (ViiV Healthcare),
ONE VIRUS, MANY DRUGS and Biktarvy (Gilead Sciences). All RT and integrase
Existing HIV drugs work to prevent T-cell infection, viral inhibitors are small molecule drugs.
replication, or the assembly and release of new viral • Disrupting viral assembly: After replication of its
particles. Examples of these strategies include: genome and production of new proteins, the final
• Preventing infection: HIV wipes out the immune step of the HIV lifecycle is the assembly of new
system because it infects a critical type of white viruses. This process depends on a specific kind
blood cell, helper T-cells. To gain entry to the T-cells, of viral protein called a protease. This enzyme
HIV binds a protein called CCR5 on the surface of processes newly-made viral proteins so that they can
those cells. Drugs that interfere with that interaction form new viral particles. Inhibiting the HIV protease
can prevent infection and are called CCR5 inhibitors. enzyme means no new viral particles. Small molecule
Currently, there is only one CCR5 inhibitor on the protease inhibitors on the market include Prezista
market—Maraviroc, a small molecule drug. CytoDyn ( Janssen Therapeutics), Tipranavir (Boehringer
(Vancouver, WA) has a monoclonal antibodyinhibitor Ingelheim), and Lexiva (GlaxoSmithKline).
of CCR5 in Phase III clinical development. The diagram below shows where in the HIV lifecycle each
of these types of drugs act.

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 These drugs have profoundly improved the lives of
people with AIDS around the world. However, the
as-yet-undiscovered cure for AIDS may arise instead
out of biotech. Check out these previous WEEKLY
articles for an overview of gene therapy and genome
editingapproaches to tackling AIDS. Biotech never rests,
and the quest for a cure pushes on.

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WORLD AIDS DAY: UNDERSTANDING HIV Life Science Training from Industry Experts

WORLD AIDS DAY: UNDERSTANDING HIV On the virus’ surface are the proteins glycoprotein
120 (GP120) and glycoprotein 41(GP41). A glycoprotein
December 1 marked World AIDS Day, an international
is simply a protein with carbohydrates attached to
day dedicated to raising awareness of the AIDS pandemic
it. This type of protein is often found on the surface
caused by the spread of human immunodeficiency virus
of viruses as well as human cells. They play a role in
(HIV) infection. According to UNAIDS, 37.9 million people
cell-cell interactions. The HIV glycoproteins enable the
globally were living with HIV at the end of 2018. Although
virus to attach to and fuse with target T-cells in order
the development of life-saving antiretroviral therapies
to initiate the infectious cycle. Other key viral proteins
has changed the disease from the death sentence it
include capsid proteins, which protect the viral RNA,
once was to a chronic disease, about one-third of those
and matrix proteins, which are thought to play a role in
infected—24.5 million—do not have access to these
viral assembly.
drugs. And for those that do have access, the treatments
come with major side effects, including fatigue, nausea, HIV is a retrovirus, meaning its genome is single-
muscle pain, and even kidney or liver damage. stranded RNA (ssRNA). For retroviruses to multiply,
they need to convert their ssRNA genome into double-
These statistics illustrate the need to improve access to
stranded DNA (dsDNA). This is accomplished by an
treatments as well as to develop effective vaccines and,
enzyme contained within the virus capsid called reverse
ultimately, a cure for the disease. This week, we cover the
transcriptase (RT). Once ssDNA has been formed from
basics of the HIV lifecycle, describing how it infects cells
RNA, cellular DNA polymerase enzymes convert it to
and makes people sick. Next week, we’ll cover how drug
double-stranded DNA (dsDNA).
discovery researchers have used their understanding
of the viral lifecycle to develop treatments that are THE VIRAL LIFECYCLE
currently on the market, and take a peek at new
Scientists have learned how HIV enters the cell,
developments in the drug discovery pipeline.
replicates and releases new viral particles. Let’s take a
HIV PRIMER look at the steps:

HIV causes acquired immunodeficiency syndrome (AIDS) • Receptor binding: Viruses can only replicate inside
because it infects and disables helper T-cells, a critical cells and so must gain entry to host cells. GP120
type of white blood cell. This incredibly destructive binds to the receptor proteins CD4 and CCR5 on the
power is carried out by just a handful of proteins surface of helper T-cells. Receptor binding selectivity
and a lipid envelope encapsulating the RNA-based accounts for the limited host range of most viruses.
genetic material. • Host cell entry (Fusion): After receptor binding, the
virus’ membrane fuses with the host cell membrane,
allowing the capsid to enter the cell.
• Release of viral RNA and reverse transcription:
The capsid shell is broken down by the cell and
the viral RNA serves a template for the viral RT to
make DNA.
• Integration of viral genome into host cell genome:
The viral dsDNA is then integrated into the host
cell DNA.
• Production of new viral proteins: The integrated
DNA produces viral RNA and viral proteins using the

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 host cell’s machinery. This is why viral infection is COCKTAIL FODDER
sometimes referred to as “hijacking” a cell.
The HIV genome is known to mutate frequently. This
• Viral assembly: New virus particles assemble with is why successful antiviral regimens involve a cocktail
the viral RNA and viral components created. Before of different drugs, and why drug developers must
leaving the cell, the viral proteins that belong on the continuously discover new drugs—as the virus mutates,
viral envelope are added at the plasma membrane resistance to different drugs emerges. The source of
and the newly-formed virus leaves the cell to infect that high mutation rate? The viral RT enzyme is “sloppy”
other host cells. meaning it makes lots of mistakes when copying the
viral genome.
By deciphering key steps in viral infection and
replication, researchers have been able to come up with
drugs to stop the virus. Join us next week to find out
how these drugs work, as well as an update on what’s in
the pipeline.

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A BREATH OF FRESH AIR FOR
CYSTIC FIBROSIS PATIENTS Life Science Training from Industry Experts

A BREATH OF FRESH AIR FOR thick mucus that characterizes the lungs of CF patients.
CYSTIC FIBROSIS PATIENTS This mucus makes them vulnerable to potentially
fatal infections.
It’s not often that a radical treatment for a chronic, often
fatal, disease comes around. The WEEKLY takes notice CF is an autosomal recessive disorder. If someone
when one does. Vertex Pharmaceuticals’ (Boston, MA) has one functioning copy of the CFTR gene, they are
innovative new drug, Trikafta, promises better health “carriers” who won’t get the disease. People with a
to a much broader range of people with cystic fibrosis copy of the malfunctioning gene from each parent, in
(CF) than previous treatments. This hereditary disease contrast, inevitably develop the illness. And while CF is
of the lungs and other mucus glands afflicts about always caused by a genetic flaw, many possible mutation
70,000 people around the globe. Passed on through combinations are associated with the disease.
one of several possible mutations in a single gene, CF The mutations come in two classes. The first group
involves the overproduction of mucus that clogs its consists of those that lessen the quantity of CFTR proteins
sufferers’ lungs and causes premature death. Without reaching the cell surface. The second group reduces the
treatment, most patients die in their 30s. CF is an orphan functioning of the proteins reaching the cell surface.
disease that is prevalent mainly in America, Europe, Drugs that work by assisting CFTR to fold correctly and
and Australia. reach the cell surface are CFTR correctors. Drugs that
Until recently, strategies for treating CF focused on enable CFTR to function correctly once it reaches the cell
the management of symptoms and involved reducing surface are CFTR potentiators.
the risk of lung infections through mucus-thinning
medications and antibiotics. Since 2012, Vertex has won
TRIKAFTA: TRIPLE CF THREAT
the approval of four different drugs—Kalydeco, Orkambi, The new CF drug, Trikafta combines three small
Symdeko, and now Trikafta—that effectively treat molecule drugs:
the underlying cause of cystic fibrosis. However, only • Kalydeco, first approved in 2012, was the first
Trikafta promises to treat a significant majority—90%— CF remedy to treat the underlying cause of the
of patients. This is because CF is caused by a number disease. Kalydeco binds to the misfolded CFTR
of different mutations, and different drugs target protein and increases its ability to remain open and
different mutations. function on cellular surfaces—a CFTR potentiator.
This WEEKLY looks at how Vertex’s game-changing new Although highly effective, the drug only works in
drug targets multiple mechanisms behind the disease. approximately 10% of CF patients. Kalydeco alone
doesn’t help patients whose mutation prevents CFTR
A PROTEIN GONE WRONG from reaching the cell surface.
CF hinges on one of several possible mutations in the • Tezacaftor and elexacafter, the other components of
gene that encodes the “cystic fibrosis transmembrane Trikafta, target the most common CF mutation. This
conductance regulator” (CFTR) protein. This protein is a single amino acid deletion in the CFTR protein
controls the production of sweat, digestive fluids and responsible for two-thirds of CF cases. This mutation
mucus. CFTR is classified as a channel protein—which results in a protein so misfolded, it never makes it to
creates a passage across the cell membrane. These the cell surface. Tezacaftor and elexacafter are CFTR
paths allow molecules which otherwise couldn’t to pass correctors. They bind to and stabilize the misfolded
through cells. In the case of CFTR, negatively-charged proteins so they can travel to the cell surface. Once
chloride ions rely on CFTR to exit cells. If CFTR doesn’t there, Kalydeco kicks in and improves the function
function correctly, the chloride ions build up inside. The of CFTR.
pileup affects the fluid balance of tissue, resulting in the

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TERM OF THE WEEK: A pharmacological chaperone is a small molecule
CHAPERONE PROTEIN drug that works on specific misfolded proteins and
encourages them to fold correctly. Tezacaftor and
Chaperone proteins assist in the correct folding and
elexacafter are pharmacological chaperones.
assembly of other proteins. Many of the proteins we
produce require chaperone proteins to ensure their The leap from simply managing CF to attacking its roots
correct molecular structure. shows the amazing potential of drug development,
highlighting biotech innovation at its best.

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VAMPIRES, ZOMBIES, PUMPKINS, & BIOTECH Life Science Training from Industry Experts

BOO-O-TECHNOLOGY! Humans also manufacture CGRP. Unfortunately,

the human peptide doesn’t work well on damaged
Vampire bats? Zombie cells? Pumpkins that heal? Yes,
endothelial cells. These make up the lining of blood
Halloween has finally encroached on the WEEKLY.
vessels. In many people with high blood pressure, this
lining is damaged.
VAMPIRE BATS
The peptide from vampire bats (vCGRP) may turn out
Earlier this year, researchers from the University of
to be superior than our own. vCGRP appears to relax
Queensland (Brisbane, Australia) published a paper
even blood vessels with compromised linings. The
on the possibility that a peptide found in the blood of
Queensland researchers say their newly-discovered
vampire bats can lower blood pressure. It’s well-known
peptide could eventually form the basis of treatments for
that the animals primarily feed on blood. In contrast to
a range of diseases, including high blood pressure, heart
previous thought, some of these bats have been known
failure, kidney disease, and burns.
to occasionally partake of human blood. That’s a subject
for another blog though.
TERM OF THE WEEK: HEMATOPHAGY
Back to peptides. What exactly are they? Peptides are
Hematophagy is the practice by certain animals of
groups of two or more amino acids, the building blocks
feeding on blood. Also called sanguinivory. Bedbugs,
of proteins. There are several peptide drugs currently
mosquitos, and leeches oh my!
on the market—Byetta (Astra Zeneca; Cambridge, U.K.),
Victoza (Novo Nordisk; Bagsvaerd, Denmark), and ZOMBIE CELLS
Trulicity (Eli Lilly; Indianapolis, IN) that help to regulate
As we age, our organs accumulate what some scientists
blood glucose levels in type 2 diabetics by activating
refer to as “zombie cells”—cells with DNA damage. They
the glucagon-like peptide-1T(GLP-1) receptor, which
no longer divide or function, but also resist dying. The
triggers the pancreas to release insulin. Other peptide
technical term for this is “senescence.” Senescent cells
drugs are in development for various types of cancer.
are found throughout our bodies. They tend to collect
The U.S. Food and Drug Administration defines a peptide
in areas that wear out, such as eyes and joints. These
therapeutic as a chain of amino acids that contain 40 or
cellular seniors secrete signaling molecules that increase
fewer components. It regulates them as small molecules
inflammation. Inflammation in turn drives several aging-
primarily because they can be synthesized in the lab.
related diseases including cancer, cardiovascular disease,
Peptides also possess some of the characteristics of
osteoarthritis, and Alzheimer’s.
large molecule drugs. They’re sensitive to digestive
enzymes and highly specific to their target diseases. Studies in mice demonstrated that clearing senescent
They’re also both delivered by injection. cells lengthened their lives by an average of 20 percent.
It also delayed organ deterioration in healthy mice.
Our body naturally produces peptides that influence
many different processes, including immunity, pain This success has inspired biotech companies to develop
regulation, and control of blood glucose. The peptide senolytics, a new class of drugs that target and destroy
isolated from vampire bats is a mutated version of senescent cells. Unity Biotechnology’s (Brisbane, CA)
human calcitonin gene-related peptide (CGRP). CGRP is a lead candidate, UBX0101, is in Phase 1 clinical testing for
vasodilator, meaning that it relaxes blood vessels. With osteoarthritis. Several other companies, including Cleara
more room to move, blood flows more easily and blood Biotech (Utrecht, Netherlands), Senolytx (Barcelona,
pressure drops. It’s all about physics. It takes more force Spain), Oisin Biotechnologies(Seattle, WA), and CellAge
to move fluid through a narrow tube (or artery) than (Glasgow, Scotland) are in the early stages of developing
through a wide one. senolytic treatments for cancer and other diseases of
old age.

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HAIRY, HEALING PUMPKIN? may speed the process of burn healing—at least in rats—
by increasing the production of collagen, a protein that
This time of year offers a panoply of pumpkin-themed
is vital for healthy tissue formation. So go ahead, enjoy
groceries: pumpkin soup, pumpkin ravioli, pumpkin spice
those pumpkin-infused treats—they just might be good
granola…the list goes on. But did you know pumpkin
for your hair and skin.
seed oil may have medicinal qualities?
One study shows that the oil may help remedy male- COCKTAIL FODDER
pattern baldness. It’s thought to block the action of Samhainophobia is the medical term for a pathological
an enzyme called 5-alpha reductase, which converts fear of Halloween. Here’s wishing all of our readers a
testosterone to DHT—the hormone that shrinks hair frightfully delightful Halloween!
follicles. Another study suggests that pumpkin seed oil

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NEW PEANUT ALLERGY TREATMENT
ON THE HORIZON Life Science Training from Industry Experts

NEW PEANUT ALLERGY TERM OF THE WEEK: HISTAMINES

TREATMENT ON THE HORIZON Histamines are small molecules produced by mast cells.
Last month, an FDA advisory committee endorsed the Once released, histamines bind to receptors on the
effectiveness of Aimmune Therapeutics’ (Brisbane, surface of blood vessels, increasing their permeability;
CA) first-of-its kind peanut allergy treatment, Palforzia. the ease with which fluid moves out of the blood vessels.
If approved, it could mean a lessening of anxiety over Histamines also bind to receptors on certain types of
the possibility of a chance exposure for the 15 million nerve cells, resulting in muscle contraction. A host of
who suffer from peanut allergies. The mere dust particle symptoms can be triggered, ranging from annoying to
of a freshly cracked peanut can be responsible for an deadly. Typical signs of histamine release include:
unpredictable cascade of reactions, including death • Increased blood vessel permeability resulting in a
brought about by anaphylaxis. runny nose and watery eyes
This WEEKLY takes a swing at explaining how allergies • Increased muscle contraction leading to throat
develop, the current treatments, and how Aimmune and constriction, difficulty breathing and swallowing
other innovative companies’ new products might change
• Extreme fluid release from tissues causing a sudden
the way allergen desensitization therapy is delivered.
drop in blood pressure, potentially bringing on a
SOMETHING TO SNEEZE AT heart attack

The host of symptoms dubbed “allergies” are the Mild allergy symptoms such as a runny nose and watery
end result of the immune system’s response to a eyes can often be successfully controlled by the use of
normally harmless substance, as if that harmless an over-the-counter antihistamine, a drug that works by
substance were a threat. An initial allergen exposure blocking the interaction of histamines with receptors on
results in the production of a class of antibodies called nerve and muscle cells. However, once anaphylaxis, a
Immunoglobulin E (IgE). A second exposure to the severe allergic reaction that includes difficulty breathing
allergen results in an “allergen-IgE antibody complex.” and heart palpitations, has occurred, it is too late for
These newly-produced complexes bind to and activate antihistamines to be effective. These symptoms can only
mast cells—a type of immune cell. As the image below be treated with an injection of the hormone epinephrine.
shows, activated mast cells send out chemical alarms in And the sooner the injection the better in cases where a
the form of histamine. life is on the line.
Epinephrine helps to reverse histamine’s effects by
decreasing blood vessel permeability, relaxing muscle
cells, and stimulating the heart. People at risk for
anaphylaxis need access to an epinephrine auto-injector,
a spring-loaded syringe that makes the lifesaving shot
readily available. This type of product is referred to as a
“combination product” because it combines a device (the
auto-injector) with a medicine (epinephrine). Mylan’s
(Canonsburg, PA) EpiPen is an epinephrine auto-injector,
as is Amedra’s (Horsham, PA) Adrenaclick.

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THE HYGIENE HYPOTHESIS five years of life. At age five, peanut allergy was assessed
and an 81 percent reduction in allergy was found in
Epidemiologists have noticed an interesting trend as
the children who regularly consumed peanuts when
countries transition from developing to developed
compared to those who avoided them.
status: an improvement in sanitation and access to
antibiotics means less pathogenic exposure and lower ALLERGENS BY THE DOSE
infection rate. As the infection rate drops, the incidence
The current allergen immunotherapy market includes
of allergies shoots up.
allergy shots (which require monitoring by a physician)
Many scientists think early exposure to infection helps and drops or tablets dissolved under the tongue (which
shift the immune response towards fighting pathogens can sometimes be taken at home). Aimmune’s Palforzia
while minimizing the production of IgE antibodies. is a capsule containing pharmaceutical grade peanut
Exposure to potential allergens (while the immune protein. The patient opens up the capsule and mixes the
system is still developing) helps to desensitize the contents with food as a means of delivery.
allergic response.
Clinical studies of Palforzia report patients becoming
THE STRATEGY BEHIND desensitized to doses at least twenty times greater
DESENSITIZATION THERAPY than the original allergy-inducing dose and in some
cases, more than 100-fold greater. The goal is to
The idea of allergy desensitization through controlled
reach a tolerance level that offers protection against
exposure has been around for decades. Desensitization
accidental eating of peanuts. The product was awarded a
is the principle behind allergy shots shown to be
Breakthrough Designation by the FDA. A final decision on
effective against pet dander, dust mites, and pollen.
FDA approval is expected by January 2020.
Desensitization therapy was once considered to be too
risky for food allergies, but a number of new studies DBV Technologies (Bagneux, France) is also in the
support the idea that gradual exposure to food allergens allergy fight. Their Viaskin skin patch delivers low
may be beneficial. doses of either peanut, milk, or dust mite allergens. By
delivering an allergen through the skin instead of the
The latest National Institutes of Health (NIH)
blood, the body will react less severely, reducing the
guidelines recommend the early introduction of peanuts
risk of anaphylaxis. The Viaskin peanut patch has been
into children’s diets, including those considered to be
awarded Breakthrough status and is currently in Phase
at high risk for developing peanut allergies because
III clinical studies.
they already have severe eczema or egg allergies.
These new guidelines are based on the results of the As allergen desensitization treatments continue to make
NIH-funded Learning Early About Peanut Allergy (LEAP) their way through the drug pipeline, allergy sufferers
study, which randomly assigned 600 high-risk infants to remain hopeful for better and easier treatment options,
either peanut-avoidance or the regular inclusion of small and, maybe even one day, a cure.
amounts of peanut products in their diet for the first

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BREATHE EASY: HYPOXIA-
INDUCIBLE FACTOR (HIF) Life Science Training from Industry Experts

EVERY BREATH YOU TAKE THE KIDNEY-ANEMIA CONNECTION

Did you know that we breathe in and out about 22,000 Anemia is a decrease in the total amount of hemoglobin
times every day? It takes that much effort to obtain in our blood. Without enough healthy red cells, our
the oxygen we need to function. Oxygen drives cellular bodies can’t get enough oxygen to our organs. A healthy
respiration, the process by which cells transform food person’s kidneys help correct this problem. When
into the energy our bodies need to do everything. But the number of red blood cells in circulation drops,
what happens to our bodies when we don’t get enough the kidneys release erythropoietin. This hormone
oxygen over time? stimulates the bone marrow to ramp up red blood cell
That’s the question that this year’s winners of the Nobel production. In chronic kidney disease (CKD), however,
Prize in Physiology or Medicine devoted much of their the body doesn’t produce sufficient erythropoietin. This
careers to answering. Last week, the Nobel Assembly causes anemia.
at Karolinska Institute (Stockholm, Sweden) awarded Our kidneys enable us to adapt to high altitude in a
the 2019 prize to William G. Kaelin (Harvard University; stepwise manner. Lower levels of oxygen stabilize
Cambridge, MA), Sir Peter Ratcliffe (Oxford University, the amount of HIF our cells make. In turn, this leads
Oxford, U.K.), and Gregg L. Semenza ( Johns Hopkins the kidneys to put out a consistently higher level of
University; Baltimore, MD) for discovering how cells erythropoietin. That increases the hemoglobin our bone
sense and adapt to differences in the amount of marrow makes, and hence boosts our blood’s ability to
available oxygen. This work increases our fundamental transport more oxygen to our cells.
understanding of human physiology. In addition, it When things are “normal,” our cells make small amounts
has important implications for the biotech industry. of HIF. It gets quickly degraded by an enzyme called HIF
Currently, several different companies are applying this prolyl-hydroxylases (HIF PHDs). Low-oxygen conditions
basic science in the quest for new ways to treat cancer inhibit the production of this enzyme, which allows HIF
and anemia. levels to rise.

TERM OF THE WEEK: HYPOXIA- Several companies are creating small molecule, orally-
INDUCIBLE FACTOR delivered drugs to block HIF PHDs and ultimately
increase the production of red blood cells under normal
The word hypoxia describes the state in which the
oxygen levels. HIF PHD inhibitors to address CKD-related
tissues in our bodies don’t get enough oxygen. Think
anemia in Phase 3 clinical development include:
about why the intrepid souls who climb Mount Everest
use supplemental air. Hypoxia can kill. Hypoxia often • Akebia’s (Cambridge, MA) vadadustat
stems from illness: lung disease, anemia, or problems • FibroGen’s (San Francisco, CA) roxadustat
with circulation. • GlaxoSmithKline’s (London, UK) daprodustat
How do healthy bodies cope with low oxygen levels?
The short answer: protein. A low-oxygen environment CUTTING OFF CANCER
triggers our cells to make a protein called hypoxia- HIF likely also contributes to angiogenesis in tumors.
inducible factor (HIF). HIF is a transcription factor—a Tumors are typically dense and lack their own blood
protein that binds to DNA in a defined location. It turns vessels, so most of them have low levels of oxygen.
on specific genes, telling them to make the protein While oxygen-deprived tumors sound doomed, they
that they code for. HIF activates genes involved in two tend not to remain that way long. The lack of oxygen
vital processes. The first is angiogenesis, the formation causes cancer cells to produce HIF. This activates the
of blood vessels. The second is the production of secretion of vascular endothelial cell growth factor
hemoglobin, oxygen-carrying red blood cells. Both assist (VEGF). VEGF triggers angiogenesis. The resulting new
in delivering more oxygen to the places that need it.

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blood vessels supply oxygen and nutrients allowing the COCKTAIL FODDER: THE
tumor to thrive. Angiogenesis may also provide a route HIF ADVANTAGE
for individual tumor cells to migrate to other parts of
You may have heard of “altitude training”—the practice
the body.
of training at high altitudes in order to increase
Some researchers are interested in developing HIF performance, especially in endurance events such as
inhibitors to thwart angiogenesis in tumors. The drugs long distance running or cycling. Altitude training works
Torisel (Pfizer; New York, NY) and Zortress (Novartis; because at elevations higher than about 5,000 feet there
Basel, Switzerland) impede a protein called mTOR, which are fewer oxygen molecules per volume of air due to
activates HIF. The anti-angiogenesis effects of the mTOR reduced atmospheric pressure. Every breath taken at
inhibiters may prevent the formation of new blood in higher elevations delivers less oxygen than it would at
tumors by inhibiting HIF. Torisel is FDA-approved for lower elevations, creating a slightly hypoxic environment
renal cell cancer. Zortress is FDA-approved for advanced inside the athlete’s cells. This hypoxia increases the
kidney cancer, metastatic pancreatic neuroendocrine levels of HIF, leading to more erythropoietin and
tumors, and hormone-positive, HER2-negative subsequent red blood cell production. The enhanced
breast cancer. oxygen-carrying capacity lasts for about ten to twenty
days after returning to lower elevations, so an athlete
who trains at a higher altitude and then competes at sea
level will have an advantage over those who complete all
of their training at sea level.

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NEOANTIGEN CANCER VACCINES Life Science Training from Industry Experts

THE ELUSIVE CANCER VACCINE characteristics. Scientists then compare tumor exomes
to those of healthy cells to ferret out differences in the
Like unicorns, the fountain of youth and calorie-free
DNA sequences.
chocolate, cancer vaccines have long proven elusive.
But the emergence of a new crop of biotech companies Not all proteins make good antigens. The most useful in
makes it clear that a cancer vaccine is no myth. The terms of cancer therapy are displayed on the cell surface,
latest advances in genomics have made it possible for where the immune system can get at them. To identify
scientists to focus on developing vaccines that train our cell surface neoantigens, researchers feed their unique-
bodies to recognize and fight an established tumor. In to-cancer DNA sequences into bioinformatics programs
this WEEKLY, we break down the science and technology which predict the probability of surface location.
of immunotherapeutic vaccines. Typically, around five percent of the mutated genes
represent potential neoantigens.
TERM OF THE WEEK: NEOANTIGEN
CUSTOM VACCINES?
An antigen is a protein (or portion thereof) that our
immune system recognizes. Think of antigens as signal The most promising neoantigens get synthesized in
flags; some flags mean “all good” and some signify “bad the lab and mixed with an adjuvant — a substance
news.” An immune response occurs when attack cells, that boosts overall immune response. Research has
such as macrophages and cytotoxic T-cells, encounter a shown that neoantigen-based vaccines with at least 20
flag that spells trouble. The best-case scenario involves different neoantigens are most likely to both prompt
our immune system recognizing, targeting and killing a strong immune response and reduce the likelihood
the cells displaying a bad antigen. In the worst cases, of resistance. The multi-pronged approach stems
problematic cells evolve strategies to remain hidden from the fact that while a tumor may mutate and stop
from our immune systems. The ability of cancer to producing one neoantigen, it’s unlikely to stop producing
“disguise” itself with good antigens makes it possible for several simultaneously.
some cancers to go undetected until very late stages. Sometimes, several different patients share common
As a tumor grows, it can accumulate additional neoantigens. In other cases, effective antigens may
mutations. Scientists have found that some mutations be unique to a single individual. Imagine a vaccine
produce new antigens which the immune system can developed for just one patient. Now that’s truly
recognize. These neoantigensare the secret sauce in personalized medicine. Such precision was unthinkable
cancer vaccines. even just a few years ago. It’s possible now because
genome sequencing has yielded tremendous efficiencies
HUNTING NEOANTIGENS in both time and money.

A precipitous rate of DNA mutation is one of the

GAME CHANGERS
hallmarks of cancer cell development. Once a tumor
establishes itself, it may produce dozens, even hundreds Many companies are already conducting clinical tests
of mutations that differentiate it from healthy cells. of neoantigen-based vaccines. Currently, it takes
Identifying specific mutations is the first step toward about six to 12 weeks to identify neoantigens and
creating a neoantigen vaccine. The whole process produce a vaccine. Industry-wide, the goal is to reduce
begins with a biopsy to collect tumor cells for study. development time to one month. Here are some of the
Thanks to advances in genome sequencing technology, key players and their products:
researchers can rapidly sequence the tumor’s entire • Marker Therapeutics (Houston, TX) has begun
exome—the portion of DNA that makes proteins. Phase II studies for a neoantigen-based vaccine (TPIV
Since only the exome produces antigens, it provides 200) for triple-negative breast cancer. This strain is
the key to revealing the neoantigens’ distinctive the most treatment-resistant type of breast cancer.

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 • Neon Therapeutics (Cambridge, MA) is testing a EASILY CONFUSED: PREVENTIVE
melanoma and glioblastoma neoantigen vaccine, VACCINE VS. THERAPEUTIC VACCINE
NEO-PV-01 in Phase Ib clinical studies in combination
The Gardasil (Merck; Kenilworth, NJ) and Cervarix
with Bristol-Myers Squibb’s (New York, NY) approved
(GlaxoSmithKline; London, U.K.) vaccines for human
checkpoint inhibitor therapy Opdivo.
papilloma virus (HPV) are often referred to as “cancer
• Gritstone Oncology (Emeryville, CA) has a vaccines.” This is because they prevent HPV infection,
neoantigen-based vaccines in Phase I/II clinical trials which is the leading cause of cervical cancer. In other
for non-small cell lung cancer. words, they are preventive vaccines. Neoantigen vaccines
The success of just one of these companies will usher in are therapeutic vaccines – vaccines given to help fight a
a new era of truly personalized medicine. specific cancer that has already emerged.

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ALZHEIMER’S DISEASE: THE WAY FORWARD Life Science Training from Industry Experts

ALZHEIMER’S DISEASE: is present at higher levels in people with Alzheimer’s

THE WAY FORWARD disease. C1q accumulates at neuronal synapses, the
key points of communication between brain cells.
Last week, we reviewed the pathology and genetics of
This protein also signals other immune cells, such as
Alzheimer’s disease (AD), and discussed the two most
macrophages—which then chomp up cellular debris
common AD drug development targets, amyloid-beta
present in affected brains. The accumulation of c1q could
plaques and tau. This week, we’ll take a look at a variety
account for the loss of synapses and accompanying
of novel AD drug targets and the companies that are
mental decline.
forging ahead to find new treatments.
South San Francisco-based Annexon is working
REVIVING THE BRAIN? on a promising therapy that centers on controlling
inflammation in the brain. ANX005, now in preclinical
Loss of neurons is Alzheimer’s signature, devastating
development, is a monoclonal antibody that mops up
effect. What if we could jump start the development of
excess c1q.
new brain cells? Two companies are trying to do just that.
Another company homing in on neuroinflammation
Neuronascent (Clarksville, MD) aims to develop small
is vTv Therapeutics (High Point, NC). Their drug,
molecule activators of neurogenesis. By screening
Azeliragon, now in Phase II clinical development, is a
large chemical libraries, the company has identified
small molecule inhibitor of the receptor for advanced
compounds that show promise of sparking neurogenesis
glycation endproducts (RAGE). RAGE is present on many
from adult neural stem cells in both tissue culture and
neurological cell types. Its activation may promote
mouse models.
amyloid-beta production and transport, tau aggregation,
The company’s lead compound, NNI-362, promoted and chronic inflammation. Preventing any of these
the growth of new hippocampal neurons in mice. developments could improve Alzheimer’s symptoms.
The new cells migrated to the correct location and
Anavex (New York, NY ) is also targeting
differentiated. Moreover, they survived long enough
neuroinflammation with their small molecule drug
to reverse previously observed cognitive declines. The
ANAVEX 2-73, and activator of the sigma-1 receptor (S1R).
hippocampus is one of the first regions of the brain
S1R is found on the surface of microglial cells, a type
to show damage in AD and is thought to play a role in
of white blood cell found in the brain. Activating S1R
memory formation and spatial navigation. Neuronascent
appears to reduce neuroinflammation. ANAVEX 2-73 is in
is preparing for Phase I trials of NNI-362.
Phase 3 clinical testing.
Neurotrope Biosciences (New York, NY) is developing
bryostatin, a drug that activates protein kinase C IMMUNOTHERAPY
epsilon (PKCꞓ). This protein plays a key role in forming
Immunotherapy—activating the patient’s immune
memories. In animal models of stroke, traumatic
system to fight disease—has taken the oncology world
brain injury, and AD, bryostatin appears to restore
by storm. South San Francisco-based Alector is trying
deficits in synapses (connections between brain cells)
this approach on Alzheimer’s. Their lead Alzheimer’s
and decrease cell death. These results suggest that
candidate, ALOO2, targets and activates the TREM2
bryostatin could help to prevent the loss of neurons and
receptor, present on immune cells active in the brain.
restore synapses.
The idea is to activate those immune cells to clear
out amyloid-beta and other potentially damaging
NEUROINFLAMMATION proteins. The TREM2 target was identified based on
Neuroinflammation is one of the drivers of genetic studies which showed that patients with a
neurodegeneration in AD, multiple sclerosis and other dysfunctional TREM2 gene had a significantly higher risk
brain disorders. Research suggests that the protein c1q of developing AD, while those whose mutations led to an

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overexpression of the receptor had a lower risk. AL002 proteasomes to fully activate in AD patients. This makes
has begun Phase 1 clinical testing. it more likely that the proteasomes will recognize and
destroy amyloid plaques and tau tangles.
SEARCHING FOR A NEW MECHANISM Oryzon Genomics (Barcelona, Spain) is taking an
Rather than target Aβ plaques directly, Yumanity epigenetic approach to Alzheimer’s. Epigenetic
Therapeutics (Cambridge, MA) is trying to identify modifications are chemical changes to gene sequences
the problems they cause. Yumanity scientists have that don’t change the information content but instead
engineered yeast cells to overproduce the Aβ protein affect how much that content is used, in other words,
and monitor its detrimental effects, such as disrupting the amount of a particular protein that the body makes.
the action of other important cellular proteins. Because Oryzon researchers identified an enzyme, lysine-specific
yeast share many molecular pathways with humans, histone demethylase 1 (LSD1), which makes epigenetic
researchers can use them to screen for potential drugs modifications to genes that results in “turning them
that address protein disruption. Promising candidates down” so they produce less of the corresponding
are then tested in Alzheimer’s patient-derived cells. By protein. LSD1 makes these changes to genes that
tackling a completely different disease mechanism, the support neuronal survival. Oryzon scientists have
new compounds may achieve greater success than seen designed a drug, ORY-2001, that inhibits LSD1. Inhibiting
so far with drugs that act directly on amyloid beta or LSD1 could mean that more neurons survive in AD
tau. Yumanity is in the lead-optimization phase of pre- patients, leading to improved cognitive function. ORY-
clinical development. 2001 is in Phase II clinical trials.
In partnership with Biogen (Cambridge, MA), It’s encouraging to know how many therapies are in the
Proteostasis Therapeutics (Cambridge, MA) is Alzheimer’s treatment pipeline. With more hard work
targeting AD-associated protein aggregates by activating and investment, perhaps one of the many introduced
proteasomes. These cellular components get rid of above will lead to a cure—or perhaps the winning
damaged proteins and dysfunctional protein aggregates therapy may include a combination of these approaches.
by dismantling their chemical bonds. The protein USP14 The world awaits a winner in this all-important race.
inhibits proteasomes. Proteostasis is working on the
preclinical development of a USP14 inhibitor that allows

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GENE THERAPY TARGETS AIDS Life Science Training from Industry Experts

GENE THERAPY TARGETS AIDS muscular atrophy. This therapy can now address other
kinds of illnesses, such as cancer and HIV. AGT is using
For many years, a diagnosis of acquired
gene therapy to eradicate latent HIV, and perhaps even
immunodeficiency syndrome (AIDS) was a death
eventually offer a cure.
sentence. It seemed unthinkable in the 1980s, but the
medical community now largely treats the disease as a CRIPPLING THE IMMUNE SYSTEM
chronic illness. This profound shift arose through the
How does HIV infect immune cells to begin with? HIV
development of highly active antiretroviral therapies
first destroys the immune system’s helper T-cells.
(HAART). These medicines inhibit the replication and
These are specialized white blood cells that recognize a
spread of the human immunodeficiency virus (HIV),
unique target, usually a specific protein on the surface
which causes AIDS. They provide hope and new life
of a pathogen. Once they lock on, the helper T’s release
to millions.
inflammatory cytokines—chemical messengers that
However, HAART comes with major side effects, such help rev up the immune system. This includes activating
as fatigue, nausea, vomiting, diarrhea, fever, muscle cytotoxic or “killer” T-cells, which directly destroy
pain, kidney and liver damage, heart disease, and invading microbes.
insulin resistance. More significantly still, the treatment
Without the helper T-cells, the immune system breaks
can never be completely effective. That’s because
down. That’s how HIV kills—by disabling the very cells
the HIV virus actually evades the treatment in two
that fight it. In fact, although HIV infects all types of
challenging ways:
helper T-cells, it appears to preferentially infect helper
• High mutation rate: The HIV genome evolves T-cells that are specific to HIV. After first wiping out those
rapidly. Its endless transformation changes the viral helper T’s, the virus can disable the rest.
proteins that HAART works on, which renders the
therapies ineffective. Nonetheless, patients try to ENGINEERING A DEFENSE
stay ahead of the virus with daily medication. In turn,
AGT’s experimental treatment, AGT103-T, is designed
pharmaceutical companies are continually working
to shore up those vulnerable helper T-cells by disabling
to develop new HIV drugs.
latent HIV and preventing new infection. The new
• Latency: People with HIV typically have a reservoir approach is a type of autologous cell therapy. That
of immune cells that contain inactive or latent HIV. means doctors engineer a patient’s own cells to use as
In this state, the virus is essentially dormant and medicine, using the following steps:
can remain so for years. However, at any time, latent
• Isolate HIV-specific helper T-cells from patient blood;
microbes can reawaken and start making more HIV.
• Use gene therapy viral vector to deliver AGT103 to
Because HAART targets only active virus, latency
the isolated cells;
enables HIV to “hide.” Many in the field of HIV research
consider tackling this viral reservoir the final frontier • Expand treated cell population in the lab and infuse
in successfully fighting the virus. This week, we focus back into patient
on technology being developed by American Gene The modified HIV-specific helper T-cells are now able
Technologies (AGT; Rockville, MD) to address chronic to disable any latent virus that reawakens, while at the
HIV infection. same time resist new infection. That means they can
effectively fight any existing HIV-infected helper T-cells.
NOT YOUR MOTHER’S GENE THERAPY The ultimate goal of the therapy is to enable HIV patients
Scientists originally conceived of gene therapy as a way to effectively fight HIV infection while eliminating
to treat disorders caused by a single genetic mutation, or reducing the need for HAART and its unwanted
such as sickle cell disease, cystic fibrosis, or spinal side effects.

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How does AGT103 make helper T-cells resistant to HIV? knocking out vif makes it impossible for the virus to
It is a type of gene therapy, only instead of delivering a replicate itself.
gene that provides the recipe for a protein, it delivers • Tat, or “trans activator of transcription,” another HIV
genes that code for “antisense” RNAs—RNAs that disrupt protein required for the virus to make new proteins.
the production of other proteins. The three proteins No tat means no new viral proteins and no new virus.
whose production is disrupted are:
AGT103 is preparing to enter clinical trials.
• CCR5, a protein on the surface of helper T-cells.
HIV needs the protein in order to get inside them. COCKTAIL FODDER: TURNING
Without CCR5, the cells will be resistant to infection. HIV AGAINST ITSELF?
• Vif, or “viral infectivity factor,” an HIV protein. The viral vector used by AGT derives from the HIV virus
If a helper T-cell is already infected with HIV, itself. Called lentiviral vectors, they’re also used to create
CAR-T therapies.

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FROM DRUG DISCOVERY TO
APPROVAL: PHASE IV Life Science Training from Industry Experts

PHARMA FINISH LINE: FDA APPROVAL The FDA’s special designations include:

Last week, we focused on the final stage of clinical Accelerated Approval allows drugs to go forward
testing, Phase III trials, where drug developers assess the using surrogate endpoints instead of clinical endpoints.
safety and efficacy of their drug in large patient groups. Surrogate endpoints, such as lowered blood pressure or
At the end of Phase III, drug developers face the moment reduced tumor size predict, rather than demonstrate,
of truth: does the study data support claims that the clinical benefits. In these cases, pharmaceutical
new drug is both safe and effective? If the answer is yes, companies must run post-market studies to verify the
then it’s time to submit either a New Drug Application anticipated effect.
(NDA) for small molecule drugs, or a Biologics Licensing Priority Review means the FDA will aim for a decision
Application (BLA) for large molecule drugs. This week, within six months.
we’ll examine the approval process, including various Fast Track is based on preclinical or clinical data that
pathways for expedited approval, and touch on post- suggests the product addresses a specified unmet
approval safety studies, also known as Phase IV. medical need. The designation enables developers to
communicate more often with the FDA. The agency
UNDER THE FDA MICROSCOPE provides guidance on clinical trial design and process,
Once a company submits either an NDA or BLA, the which helps resolve questions or issues quickly. These
FDA takes about a year for review, which seems designees also qualify for Accelerated Approval, Priority
governmentally slow unless you consider that most Review, and Rolling Review—which allow developers
applications run 100,000 pages or longer. There are to submit each section of an NDA or BLA as they finish,
three possible responses: rather than all at once.
• Approval letter: Ta dah! Breakthrough Therapy designates drugs that may
• Approvable letter: Close, and this is how to get the greatly improve patient health. The bar is set high
cigar! FDA requests correction of minor deficiencies, to join this privileged group, though. It requires
labeling changes, or post-approval studies. preliminary clinical evidence of effectiveness. Once
granted, Breakthrough designees receive Fast Track
• Not approvable letter: Thanks for playing! FDA
advantages, as well as intensive guidance on their
elaborates on deficiencies in the application and why
development program as early as Phase I. They also get
the drug is not approved.
an “organizational commitment involving senior FDA
For many drugs, the development, testing, and
managers,” according to the FDA website.
approval journey is straightforward—difficult,
but straightforward. Orphan Drugs are often found in the above categories.
Companies develop them for rare diseases, those which
THAT SPECIAL SOMETHING affect fewer than 200,000 Americans. These include
hemophilia and Gaucher’s Disease, a genetic disorder
Certain drugs are given special consideration throughout
that causes skeletal and neurological issues. Prior to
the FDA approval process. These particular therapies are
the Orphan Drug Act of 1983, the industry had no
eligible for regulatory designations that speed up the
financial incentives to work on therapies for such small
review and get a product to market more quickly. They
populations. Orphans lacked commercial sponsors,
must first meet two criteria. A drug must target a serious
“parents,” to shepherd them through lengthy and
condition that likely results in death or significantly
expensive trials. The orphan drug legislation provides
impairs daily living. Examples include cancer and
the following incentives:
Alzheimer’s disease. Secondly, the drug also needs to
address a major, unmet medical need. That is, either no • Federal tax credits of up to 50% off research costs
medicine exists, or current therapy has safety issues. • Increased protection from generic competition

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 • Waivers of FDA application and product fees quickly reveal whether a patient can benefit from Plavix.
(amounting to hundreds of thousands of dollars) This unlooked-for result spurred the FDA to require that
the Plavix label carry a black box warning—a warning
COCKTAIL FODDER: PDUFA outlined in a black box, which is the strictest warning put
The Prescription Drug User Fee Act (PDUFA) requires on the labeling of prescription drugs or drug products by
drug developers to pay a fee to the FDA to help fund the the FDA.
work necessary for timely approvals. The “PDUFA date” The rigorous path from preclinical testing to approval
of a drug is the date by which the FDA has committed to usually takes a decade or longer, with no promise of
review its application. For 2020, the PDUFA fee required success. According to the Biotechnology Innovation
when submitting an NDA or BLA that requires clinical Organization, approximately one in ten drugs entering
data is $2,942,965. clinical testing ultimately make the grade. The highest
rate of failure occurs in Phase II; only 31% of drugs
FINISHED, NOT DONE proceed to Phase III. Of those that make it to the final
Phase IV generally refers to post-market studies, which Phase III, a little more than half—58%—are considered
companies undertake after a drug is approved and at the safe and effective enough for an NDA or BLA submission.
pharmacy. Drugs that were approved using a surrogate From start to finish, only 10% of the drugs that begin
endpoint are monitored to confirm clinical efficacy. For Phase I reach the market.
all drugs, safety is monitored and confirmed. Phase IV
includes informal studies of doctor and patient reports,
which sometimes reveal unanticipated side effects.
Such “surprises” aren’t uncommon in brand new drugs,
because so many more people take them once out
on the market versus the small number of patients in
clinical trials.
For instance, after the anti-clotting medicine Plavix
(Bristol-Myers Squibb; New York, NY) was approved,
doctors found it less effective in some people than
expected. Further probing found that the drug wasn’t
fully activated in about 14% of patients because their
Despite the low odds, innovative companies continue to
bodies produced a less active form of the liver enzyme
bring new drugs to market every year. Stay with us as we
that activates Plavix. Fortunately, a genetic test can
continue following their stories.

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FROM DRUG DISCOVERY TO
APPROVAL: PHASE III Life Science Training from Industry Experts

PHASE III IS NO GUARANTEE A crossover study’s strength lies in its ability to capture
differences in response to a drug versus a placebo in the
Our last Biotech Primer WEEKLY explored the riskiest
same patient. This eliminates the inevitable individual
part of the human clinical trials pathway: Phase II.
variations among subjects in the experimental and
About 70% of drugs that enter Phase II never make it
control groups in the more traditional parallel study.
out. Most often, it’s because they fail to demonstrate
effectiveness. Even making it to Phase III is no guarantee
of success. About 40% of drugs fizzle out during this
period. In today’s WEEKLY, we look at the final stage of
clinical testing and the innovative clinical designs that are
pushing for faster drug approvals.

CHOICES, CHOICES, CHOICES

Phase III clinical trials continue to test a treatment’s
efficacy and safety, but in larger groups of patients.
Bigger groups mean more statistically significant
results. As in Phase II, the trials have traditionally been
randomized, double-blind studies. If the investigational
drug appears to work, patients are allowed to continue THE NEW KID IN TOWN:
taking the medication after the trial ends, before ADAPTIVE DESIGN
regulatory approval. Adaptive studies offer more flexibility than traditional
In addition to randomized, double-blind trials, designs, and are gaining in popularity as more
researchers have other study designs at their disposal, efficient in bringing new drugs to the market. They
which we will explore below. allow investigators to modify the trial design as they
go, rather than spending time and money pursuing
THE PATRIARCHS: PARALLEL drug formulations or dosages that ultimately prove
VS CROSSOVER DESIGNS ineffective. For example, researchers may separate
participants into different dosage groups. At a
Parallel studies use the classic experimental design.
prespecified time, they may note how patients respond
Participants are divided into two groups. The treatment
to different doses. If one dose seems more effective, the
group receives the experimental drug. The control group
researchers will conduct the rest of the trials using only
receives either the current standard of care or a placebo.
that dosage. Researchers set the trial protocol, which
Investigators sometimes decide to conduct a crossover lays out the adaptation schedule and processes before
study. Here, both groups receive the medicine and then the trial begins.
the placebo or vice versa. The treatments are separated
One prominent example of adaptive design is I-SPY 2
by a washout period in which patients are taken off the
study (Investigation of Serial Studies to Predict Your
study medicine (or placebo) to eliminate any effects.
Therapeutic Response with Imaging and Molecular
Both parallel and crossover studies include a baseline Analysis). This trial, being conducted by 20 different
period before patients take the drug or placebo. It allows cancer research centers across the US, examines up to
researchers to gather the initial health information 12 different breast cancer therapies at once. Drugs that
against which they will compare changes observed show promise proceed to Phase III, allowing space for
during or after the study. For example, when assessing other drugs to enter the trial. “Graduates” of the I-SPY 2
a new cholesterol medication, researchers might take study include AbbVie’s (North Chicago, IL) Veliparib and
baseline cholesterol levels. Merck’s (Kenilworth, NJ) MK-2206.

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The 21st Century Cures Act promotes the use of NEW APPROACH TO FIGHTING
adaptive trial design. This benefits volunteers, as CANCER: UMBRELLA DESIGN
researchers assign more patients to study groups taking
These trials reflect the fact that scientists classify most
medicine that demonstrates promise. It’s good news for
cancers as specific subtypes based on the mutation
non-study patients too, who stand to benefit from new
involved. An umbrella study may look at patients with
treatments getting to market faster.
one type of cancer, for example, lung cancer. However,
NEW APPROACH TO FIGHTING researchers divide participants into subgroups based
CANCER: BASKET DESIGN on the particular mutation behind their lung cancer, and
treat them with drugs designed to target the subtype
The design of clinical trials for new oncology drugs is
specifically. This allows researchers to identify patients
changing to reflect our increasing understanding of
most likely to benefit from the new drug.
cancer as a disease driven by mutations. Traditionally,
trials test drugs on a specific illness, say, lung or One ongoing umbrella study is the National Cancer
breast cancer. In contrast, basket trials classify cancers Institute’s Alchemist Lung Cancer Enrichment Marker
according to the mutation they exhibit, not the tissue Identification and Sequencing Trials (ALCHEMIST) study.
they affect. For example, the FDA initially approved the In it, researchers sequence patients’ tumors and assign
drug Zelboraf for melanoma patients with a mutation them to different treatment groups according to the
in their “BRAF” gene. Researchers at Memorial Sloan subtype mutation.
Kettering Cancer Center (New York, NY) explored
whether Zelboraf could work against other cancers with
the same genetic signature, the BRAF mutation. The
results indicated that Zelboraf also effectively treated
BRAF-mutated non-small cell lung cancer, Erdheim-
Chester disease, and Langerhans cell histiocytosis. These
last two, rare blood cancers, illustrate a key benefit of
basket trials: they may offer patients with rare cancers
better access to clinical trials, depending on their tumor’s
genetic signature.

Phase III is the final hurdle for drug candidates. If a

drug “survives,” things are looking good for developers
and patients alike. It’s not a slam dunk yet though, so
next week we look at the approval process and what
comes next.

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FROM DRUG DISCOVERY TO
APPROVAL: PHASE I/II Life Science Training from Industry Experts

PHASE I AND II CLINICAL TRIALS This first stage involves a number of different tests.
First, volunteers take escalating doses of the drug under
Every drug in clinical use today, from the latest CAR-T
close observation. If and when they experience adverse
treatment to older cholesterol-lowering statins,
effects, they stop taking the drug. This establishes the
share one thing in common: they have all successfully
Maximum Tolerated Dose, or MTD, which becomes a
navigated the rigorous clinical trials process. This is no
benchmark for the remaining trials. The MTD helps
small feat, as only ~10% of the drugs that enter Phase I
assure investigators and subjects that the treatment is
testing successfully emerge as marketed products. Those
unlikely to be toxic.
few drugs that show remarkable success in early clinical
trials make headlines, and deservedly so. This week, we’ll Other studies look at pharmocodynamics (PD) and
take a look at the first two phases of clinical trials. pharmacokinetics (PK). The first examines what the drug
does to the body; the second, what the body does to the
TERM OF THE WEEK: ENDPOINT drug. These investigations help determine drug dosage.
Variations in how people of different sizes, ages, and
Clinical trials measure endpoints, that is, major health
genetic backgrounds etc., will likely respond to a new
outcomes. There are generally two types:
drug make testing in different populations critical.
• Clinical endpoints refer to benefits such as
survival, decreased pain, the absence of disease, or PHASE II
greater mobility.
Phase II examines drug efficacy as well as continuing
• Surrogate endpoints substitute for clinical safety tests. Phase II trials involve larger groups of
endpoints when they are impossible or impractical participants, all of whom are patients. Group size varies,
to measure. For instance, a clinical benefit, such depending on the target market. A drug being developed
as survival, hopefully, takes decades to observe. for Type 2 diabetes needs far more participants than one
Researchers may instead look to shorter-term for a rare disease such as ALS.
phenomena. For example, in studying a drug
Phase II studies are usually randomized, double-
designed to prevent heart disease, they can monitor
blind studies. This means that patients are randomly
cholesterol levels instead of decreased fatality from
assigned the drug or the placebo—a substance that
heart attacks. Similarly, in some cancer treatments,
has no therapeutic effect—and even the researchers
reduced tumor size stands in for longer life.
involved don’t know who receives which. This helps
The FDA requires that clinical protocols clearly define guard against bias in determining who goes into which
endpoints. They are front and center in the application group. Otherwise, investigators may naturally want
companies submit to the FDA that request permission to ensure the sickest patients receive the promising
to study a new drug. This application is called an experimental treatments.
Investigational New Drug application or IND.
If there is already an effective drug on the market, the
standard of care, patients who are not receiving the
PHASE I experimental drug will receive the standard of care.
The FDA divides clinical studies into three main phases. According to the National Institutes of Health (NIH),
Phase I usually tests drug safety in healthy volunteers, the standard of care is a treatment accepted by medical
typically one hundred or fewer. In some cases, Phase I experts as proper for a certain disease and widely used
trials may use patients rather than healthy volunteers. by medical professionals. For example, in clinical trials
For example, cancer drugs have a level of toxicity that we for Amgen’s (Thousand Oaks, CA) new cholesterol
would not want to expose healthy volunteers to, but that drug Repatha, researchers compared its safety and
toxicity is an acceptable risk for patients who may have efficacy against statins, the current standard of care for
no other options. cholesterol reduction. Repatha was demonstrated to be

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as effective as statins, and had a better safety profile in COCKTAIL FODDER
some patients, which led the FDA to approve Repatha.
The first antibiotic, penicillin, was discovered in 1928
THE POWER OF POSITIVE THINKING when microbiologist Alexander Fleming noticed the mold
Penicillium secreted a substance that killed his bacterial
Investigators measure drug efficacy by monitoring
cultures. Inspired by this story, biochemist Akira Endo,
predefined endpoints. They always look at effectiveness
who was working at Sankyo Pharmaceuticals (Tokyo)
relative to a control group, people who receive either
in 1972, decided to screen Penicillium cultures in hopes
standard of care treatment or a placebo.
of finding a chemical that would inhibit cholesterol.
The “placebo effect” may stem from an expectation or Endo’s quest paid off. He discovered a compound that
belief that the “medicine” will work. The placebo control eventually went on to be one for the first statin drugs
is very important since scientists have firmly established approved in 1987.
that placebos can significantly affect patient health.
Why would mold cells secrete an anti-cholesterol drug?
Recent evidence shows that people dealing with chronic
For the same reason they secrete penicillin—to kill the
pain can produce natural opioid painkillers in response
bacteria that they are competing with for resources.
to a placebo. More rarely, the reverse of the placebo
Cholesterol is required for bacteria to build functional
effect—the “nocebo effect”—occurs. In other words,
cell walls.
some patients expect a study drug will either not help or,
in fact, harm them. Next week, we’ll continue along the pathway to approval
as we zoom in on drug development Phase III and IV.

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THE MANY FACES OF BOTOX Life Science Training from Industry Experts

THE MANY FACES OF BOTOX received Botox injections noticed a welcome side effect:
fewer wrinkles between their eyes. By temporarily
Botox. This powerful protein has become somewhat
paralyzing the “frown” muscles, Botox made the
of a meme for people of a certain age wanting to look
unwanted furrow disappear. The FDA officially approved
less than that age. Botox’s applications, however, go far
Botox for cosmetic use in 2002.
beyond the cosmetic, as recent industry headlines about
AbbVie’s (North Chicago, IL) acquisition of Botox maker, Since then, the FDA has approved many other
Allergan (Irvine, CA) remind us. This week, we explore indications: overactive bladder syndrome, chronic
the science of Botox and its many medical uses. migraine, upper and lower limb spasticity, cervical
dystonia, eyelid spasms, and excessive sweating.
PICK YOUR POISON
POISON WITH A HEART
Botox is a neurotoxin—a substance that destroys nerve
tissue. It’s produced by the bacterium Clostridium Atrial fibrillation—an abnormal heart rhythm—can result
botulinum and causes botulism, a rare but sometimes from heart surgery. Researchers are finding that Botox
deadly illness. Botulinum, (Botox’s formal name) may have yet another important role to play here as well.
interferes with key muscles in the body. This results in Preliminary studies suggest that injecting the neurotoxin
paralysis and even death. People contract food-borne into the nerve-rich fatty pads of tissue outside the
botulism typically by eating home-canned food that heart reduces the rate of atrial fibrillation and related
has been prepared incorrectly. Botulism toxin is one of complications in cardiac patients. Presumably the
nature’s most lethal toxins. It’s twice as potent as the benefit comes from temporarily blocking nerve signals
tetanus toxin and 100,000 times as potent as the poison to the heart. Larger scale studies will determine whether
gas sarin. this should become standard treatment.

How in the world did such a deadly substance become OTHER TWOFERS (THREEFERS…
a medicine? It all comes down to dose. The amount of FOURFERS…)
toxin used for medical or cosmetic purposes is miniscule.
Botox is just one of many drugs developed for one
It’s estimated that a baby-aspirin-sized amount of the
condition only to yield unexpected second (or third, or
powdered toxin is enough to provide the global supply of
fourth…) applications. Some of the best known include:
Botox for a year.
• Lumigen (Allergan) was originally approved for
Just how does Botox paralyze muscles? Botox works by
glaucoma. Patients and physicians noticed that it
blocking acetylcholine. This neurotransmitter is released
also resulted in long, lush eyelash growth. A version
by motor neurons to activate muscles. Putting the
of the drug for cosmetic purposes sells under the
brakes on acetylcholine puts the brakes on our muscles.
brand name Latisse.
Unchecked, the toxin can kill by freezing the diaphragm
muscles, leading to respiratory failure. • Proscar (Merck; Kenilworth, NJ ) was initially
approved to treat enlarged prostrate, but came
POISON WITH A PURPOSE with the happy side effect of decreasing male-
In very small, localized doses, this paralyzing toxin can pattern baldness. Propecia is the version marketed
actually benefit human health. The FDA first approved for baldness.
the toxin in 1989 to treat strabismus, otherwise known • And of course, Pfizer’s (New York, NY) Viagra was
as cross-eyes. Strabismus is caused by overactivity originally being developed to treat high blood
in one of the two muscles responsible for lateral eye pressure, an indication which proved unsuccessful.
movement. Relaxing the muscle results in normal eye But, oh what a failure!
alignment. Some “mature” strabismus patients who

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COCKTAIL FODDER • Prialt, an analgesic for severe and chronic pain,
comes from a toxin produced by the cone snail.
Poisonous animals also provide a seemingly dangerous
Approved in 2004, it was developed by Elan
source of lifesaving remedies. Again, the dose makes the
Pharmaceuticals, now Jazz Pharmaceuticals (Dublin,
difference, as do harvest methods! Examples include:
Ireland).
• Captopril, a high blood pressure medicine, is based
• Integrilin, a powerful anti-clotting drug, derives from
on pit viper toxin. It was developed by Bristol-Myers
the venom of southeastern pygmy rattlesnakes. It
Squibb (New York, NY) and approved by the FDA
was developed by Millennium Pharmaceuticals
in 1984.
(Cambridge, MA) and approved in 1998.
• Another gift from the pit viper is Defibrase, which
• Byetta helps control blood glucose levels in type
can stop bleeding. It comes from a purified version
2 diabetics and is a synthetic version of a protein
of a protein found in the snake’s venom. It was
obtained from Gila monster venom. It was developed
developed by Pentapharm (Basel, Switzerland) and
by Amylin Pharmaceuticals(San Diego, CA) and
has been approved in Japan.
approved in 2005.

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KEEPING CANCER IN THE CROSSHAIRS Life Science Training from Industry Experts

KEEPING CANCER IN THE CROSSHAIRS For the phenotype approach, researchers test a chemical
WITH TARGETED THERAPIES compound in cell, tissue, or animal models to see
whether it exerts the desired effect, such as putting the
One of the biggest stories in last week’s biotech news
brakes on cancer cell division. Compounds that seem
headlines was Pfizer’s (New York, NY) acquisition of
beneficial move forward in the drug discovery process.
Array Biopharma (Boulder, CO) for the tidy sum of $11
The mechanism of action—the drug target—sometimes
billion. Array specializes in targeted cancer therapeutics.
remains undefined until much later on.
The company is known both for developing its own
therapies and its discovery platform. The platform has BRAFTOVI/MEKTOVI
enabled Array to partner with a host of other companies
Using its discovery platforms, Array Biopharma gained
seeking to discover and develop new drugs. Because of
FDA approval for two kinase inhibitors, Braftovi and
its very specific expertise, Pfizer plans to keep Array’s
Mektovi. The drugs were approved to use together to
Boulder facilities open as a research site for Pfizer.
treat melanoma caused by specific mutations in the
This WEEKLY takes a closer look at targeted therapeutics BRAF gene. These mutations show up in about half of
and their discovery platforms. all melanomas.

TERM OF THE WEEK: SHAPE-SHIFTERS TO THE RESCUE

TARGETED THERAPY
What are kinase enzymes? Further, how does inhibiting
Targeted therapy has improved the lives of countless them help people with cancer? Kinases are enzymes
cancer patients and their loved ones. These specialized that add a phosphate molecule to proteins. Phosphate
treatments identify and attack specific kinds of cancer molecules are small, negatively-charged chemical
cells while causing less harm to healthy cells than groups. The addition changes the shape of the recipient
chemotherapies and other more traditional approaches. protein. Mostly, this shape-shifting turns the protein “on”
To the average person, targeted therapies can almost from an “off” state. This process, phosphorylation, plays
sound like magic. an important role in sending growth signals within a cell.
Behind the magic, science is working hard. Targeted Typically, kinase enzymes transmit growth signals only
therapies block the action of certain enzymes, proteins, when activated by a growth factor. However, mutations
or other molecules that fuel the growth and spread of cause some of them, like BRAF, to stay active all the time.
cancer cells. This heightened state triggers excessive cellular growth
and division: cancer. Inhibiting the mutant kinases can
DIFFERENT PATHS TO BULLSEYE shut cancer down.
How do we even get to a targeted therapy in the first
Braftovi halts the mutated, overactive version
place? Typically, there are two strategies: target-based
of the kinase BRAF. Mektovi inhibits the kinase
and phenotype-based.
enzyme MEK, which is overactive in many cancers,
In target-based drug discovery, researchers home in on including melanoma.
one molecule integral to a disease process. For instance,
some cancers involve a cellular enzyme that mutates TARGETED THERAPIES’ ACHILLE’S HEEL
to promote the wrong kind of cell division. Researchers Although hailed as safer and more effective in treating
screen the enzyme against enormous chemical cancer, targeted therapies have a critical limitation:
libraries to see what comes up. They’re searching for a resistance. People may develop resistance to their
molecule that interacts with the drug target in a possibly treatments in two ways:
therapeutic way. Companies then further screen any
• The target protein mutates in a way that prevents
“hits” to find potential drugs.
the therapy from interacting with it further.

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 • The tumor finds a different way to grow, one that no Ongoing clinical studies are examining combinations of
longer depends on the target molecule. targeted therapies and immune checkpoint inhibitors.
To reduce the likelihood of resistance, doctors often Targeted therapies continue to be an important weapon
prescribe targeted therapeutics with other drugs. This in the fight against cancer. As researchers continue to
is why the FDA approved Braftovi to be prescribed in collect and characterize more and more tumor genome
combination with Mektovi, which inhibits a different sequence data, which aids in identifying new targets, we
cell-signaling pathway. Clinical studies demonstrate that expect to see additional new targeted therapies.
the combination is more effective than Braftovi alone. We’ll be off next Thursday to celebrate Independence
Day. Enjoy the holiday and we’ll be back in two weeks!

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NEW GENE THERAPY DRUG EXPLAINED Life Science Training from Industry Experts

NEW GENE THERAPY DRUG EXPLAINED • Type 1: The most severe and the most common type
of SMA. Symptoms are usually present within the
Novartis’ (Basel, Switzerland) Zolgensma grabbed
first few months of life, and these babies often do
headlines last week as it garnered the second FDA
not display movement of any kind. As the disease
approval of a gene therapy treatment for an inherited
progresses, toddlers have trouble with swallowing
disease. Designed to treat spinal muscular atrophy
and respiratory function. SMA Type 1 is usually fatal
(SMA), Zolgensa is also set to be the most expensive drug
by age two.
ever approved, with the one-time treatment ringing up at
$2 million. In this WEEKLY, we’ll take explain what SMA is • Type 2: Symptoms manifest between six and
and how Zolgensma treats it. 18 months. These children can typically sit but
not stand or walk. Respiratory function is often
SMA PRIMER compromised and is a major concern, however with
the help of machines many of these patients live
Our nervous system consists of the brain, spinal cord,
into adulthood.
and a vast network of nerves that feed into every tissue
of the body. Motor neurons are a type of nerve cell • Type 3: Symptoms occur after age one. These
that sends messages from the spinal cord to muscles, patients are usually able to walk, but may lose
enabling movement. that ability as the disease progresses. Respiratory
function is less impaired, and life expectancy is often
In order for the motor neurons to do their job, a
near normal.
functional protein called the survival motor neuron
(SMN) protein is necessary. The survival motor neuron • Type 4: This is the adult-onset form, typically
1 (SMN1) gene is responsible for producing most of the manifesting at age 30 or later. Muscles gradually
SMN protein used by the body. A second, closely related weaken, and the patient often needs to use
gene is the survival motor neuron 2 (SMN2) gene, which a wheelchair later in life. Life expectancy is
produces a much smaller amount of SMN protein and is not affected.
seen as a sort of “back-up” version to SMN1. Zolgensma was approved to treat SMA Type 1, the most
SMA is caused by a variety of mutations in the SMN1 common—about 60 percent of cases—and severe form
gene. Without functional SMN protein, the neurons do of the disease.
not work correctly and eventually die. How soon they
die depends on the extent of the SMN deficiency, which GENE THERAPY FOR SMA
correlates with the severity of the disease: the less SMN As a single gene disorder, SMA is an ideal candidate for
produced, the more severe the disease. a gene therapy approach because delivering a “good”
The back-up gene, SMN2, produces a small amount of copy of the mutated gene should cure the disease
functional SMN protein. However, differences in the way by supplying a permanent copy of the correct SMN1
SMN2 functions means most (but not all) of the protein protein-making instructions.
is non-functional and degrades shortly after being Using a “vector”—a virus stripped of its disease-causing
produced. Patients with less severe forms of the disease ability—scientists are able to safely deliver corrected
usually have extra SMN2 copies because ultimately, genes into targeted cells. In the case of SMA Type 1, the
even tiny amounts of SMN protein provide some motor AAV9 vector crosses the blood-brain barrier and delivers
nerve function. corrected copies of the SMN1 gene into motor neuron
An orphan disease, SMA affects about 1 in 10,000 babies cells in the brain.
born in the United States. The four generally accepted In clinical trials, babies who received Novartis’ Zolgensma
classifications of SMA are: showed marked increases in SMN production and in
movement—with most participants even talking and

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sitting without assistance. These results, combined with I will also be teaching a full-day class, BioBriefing, on
its status as a one-and-done cure for this devastating Sunday, June 2 during BIO2019. This class explains the
disease, mean that all the hype surrounding this science driving today’s hottest biopharma innovations,
breakthrough-designated drug just might be justified. and is ideal for anyone who works on the business side
of pharma without a science background. There is a cost
SEE YOU AT BIO2019! to attend and registration is required – register today!
I will be giving a 75-minute talk next week on the Science Interested in learning more about how drugs are priced?
Behind Immunotherapies at the BIO International Join Dr. Shane Desselle for his one-day Understanding
Convention in Philadelphia. Please join me on June Drug Pricing class on Sunday, June 2 during BIO2019.
3 at 2:30 pm, 106AB, Level 100 in in the Philadelphia Learn how pharmacoepidemiology influences drug
Convention Center. In this talk, we’ll review the basics reimbursement policy in the U.S. and how this affects
of how the immune system works, then explore how pharmacoeconomics. There is a cost to attend and
new treatments use patients’ immune system to fight registration is required – register today!
diseases such as cancer. The talk is free for all conference
And of course, please be sure to come visit Biotech
access attendees.
Primer at booth 3814. Say hi, grab a T-shirt, and enter
our raffle to win a pair of AirPods. See you in Philly!

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VIRUSES TO THE RESCUE? Life Science Training from Industry Experts

VIRUSES TO THE RESCUE? a standardized way. Here are some biotech companies
delving into the promising world of bacteriophage-
Last week, a paper in Nature Medicine described a British
based therapeutics.
teenager whom doctors pulled back from the brink
of death. The young woman had developed a deadly, THE VIROLOGIST-
antibiotic-resistant infection following lung surgery. The MIXOLOGISTS GET BUSY
treatment? Phage therapy, in which viruses are used
The first multicenter clinical trial examining
to kill dangerous bacteria. This novel approach made
bacteriophage as antibacterial treatments was
headlines around the world. Let’s look at the science
initiated in 2015 by French biotech Pherecydes (Paris,
behind phage therapy for antibiotic-resistant bacteria.
France). Since bacteriophage are an entirely new type
TERM OF THE WEEK: BACTERIOPHAGE of biologic drug, the researchers had to establish
production protocols to meet good manufacturing
A bacteriophage—also referred to as a phage—is a
practice guidelines.
virus that infects bacteria. By attaching to the microbe’s
surface, a phage punches holes in the membrane The scientists are studying two viral “cocktails”—
and injects its own genetic material. The intruding mixtures of different bacteriophage that have shown
phage replicates, creating so much new virus that the activity against different substrains of a particular
bacterium explodes. A slew of fresh viruses infect other bacteria. The first contains 13 phages targeting P.
bacteria, wiping out the population. aeruginosa; the second, 12 phages that target E. coli.
The company is evaluating them against burn wound-
The word “bacteriophage” comes from the Greek word
associated infections. The company is also on the cusp
phagein—“to devour.” These ravenous microbes often
of clinical trials of phage therapy to treat diabetic foot
have a taste for the bacteria that can kill us.
ulcers infected by S. aureus.
Typically a phage ”partners” with only one type of
Other companies testing phage cocktails include:
bacteria. They’ve coevolved for millennia, each adapting
and changing in response to the other. That’s key. It • AmpliPhi Biosciences (Richmond, VA): Phage
means that we’re much less likely to develop “phage cocktail AP-SA01 is now in Phase II clinical testing
resistance”—as we have to many antibiotics. for antibiotic resistant S. aureus against chronic
rhinosinusitis and acute, chronic wound and skin
So when researchers tweak a bacteriophage for
infections. Additional targets include bacteremia,
therapeutic use, they select one that will attack only
endocarditis, prosthetic joint infections,
nasty bacteria. This remarkable precision leaves many
osteomyelitis, and diabetic foot ulcers.
strains of “friendly” bacteria that comprise our gut
microbiome alone. Humans have safely coexisted with • TechnoPhage (Lisbon, Portugal): Phage cocktail TP-
bacteriophage literally for ages, which suggests the 102 is in Phase I studies for bacteria associated with
viruses pose only a minimal safety risk. chronic ulcers.

We’ve known about the bacteria-devouring ability of • Intralytix (Baltimore, MD): Completed Phase
phage for about a century. However, the seemingly I studies of a bacteriophage to treat infected
miraculous advent of antibiotics in the late 1920s wounds. The company is also developing phage-
shifted medicine’s focus to the new wonder drugs. based biologics to fight food-borne pathogens. Its
Antibiotics were much easier to manufacture and test in EcoShield targets coli, and SalmoFresh, salmonella.
controlled settings. • EpiBiome (South San Francisco, CA): Phage cocktails
The alarming emergence of antibiotic resistance has targeting E. coli and S. dysenteriae diarrheal infections
renewed interest in these killer viruses, which are for are in preclinical development.
the first time starting to be manufactured and tested in

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THE BIOENGINEERS TAKE A WHACK that essentially chew holes in the bacterial cell wall.
ContraFect (Yonkers, NY) is conducting Phase II clinical
Creating a phage cocktail is challenging. Happily, it’s also
studies of its drug CF-301—a lysin—to treat S. aureus
possible to genetically engineer a synthetic virus that
bloodstream infections.
combines the desired properties of multiple phages into
a single genome. SNIP SNIP!
For example, scientists can insert genes into a phage Locus Biosciences (Morrisville, NC) identifies
genome that increases the range of bacteria subtypes bacteriophage that target specific strains of bacteria and
an individual phage can attack, while maintaining the engineers them to deliver CRISPR components to those
specificity that prevents it from turning on benign bacteria, including guide RNA and the Cas3 protein.
bacteria. Researchers could also add genes to further The guide RNA directs the Cas3 protein to cut up the
amplify the bacteriophage’s antibacterial response. bacterial DNA.
Companies with engineered bacteriophage in preclinical
For CRISPR genome editing, the Cas9 protein is used.
development include Synthetic Genomics (San Diego,
Cas9 cuts target DNA in a single, precise location. Cas3
CA) and EnBiotix (Cambridge, MA).
chews up target DNA much more thoroughly than Cas9,
LYSINS IN WAIT making it a better choice for destroying a bacterium.
Locus plans to enter clinical trials in 2019 with phage
A third approach to harnessing the therapeutic power
treatment for urinary tract E. coli infections.
of bacteriophage lies in isolating what makes them
toxic. For example, to inject their genome into bacteria, Although still in its early days, bacteriophage therapy
phage must essentially bash holes in their membranes. offers the possibility of a safe, effective and intriguing
This blow is itself very damaging to bacteria. The viral solution to one of the major public health crises of
protein that creates these tears are lysins—enzymes our time.

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COMING SOON? A NEW BIOLOGIC Life Science Training from Industry Experts

COMING SOON? A NEW BIOLOGIC

For the past two decades, monoclonal antibodies (mAbs)
have dominated the biologic drugs market. Doctors
use these blockbuster products for a range of diseases,
including many cancers and autoimmune diseases. In
addition to their effectiveness, mAbs generally produce
fewer side effects than traditional small molecule drugs.
In the world of pharmaceuticals, biologics have “high
specificity.” In other words, they interact only with their
intended target. That’s the good news about mAbs.
Despite their therapeutic success, mAbs are not perfect.
Their complex structure means that they must be The secondary level consists of highly regular, local
produced in Chinese hamster ovary (CHO) cells rather three-dimensional structures within a protein.
than bacterial cells, which is significantly more expensive Secondary structure comes from interactions between
due to the higher cost of CHO cell growth media as neighboring amino acids. Two common types are alpha
well as the longer time required for CHO cells to grow. helices and beta sheets. Regions of secondary structure
Monoclonal antibodies are also very large, molecularly may group together to form a motif—a pattern that is
speaking—which means that in some instances, they are found in several different proteins.
not effectively able to penetrate their target tissue. Alpha-helices and beta-sheets interact to form a complex
This week, we examine an up-and-coming alternative to three-dimensional structure known as tertiary structure,
the amazing, somewhat problematic mAb: DARPins— but it is the secondary structures known as alpha helices
Designed Ankyrin Repeat Proteins. This new type of that form the basis for the newest implement getting
therapy is currently in clinical development and seems ready to emerge from the pharmaceutical toolbox—
to possess many of the advantages of mAbs without the the DARPin.
complexity and size. Here’s how.

DISASSEMBLING THE PROTEIN PUZZLE

To understand a DARPin, we need to first think about
how proteins are built. Proteins are chains of amino
acids—complex, three-dimensional forms that arise
from intermediate layers of structure. The most basic
level of protein structure is the primary structure, a
linear sequence of amino acids that make up the protein.

DESIGNER PROTEINS
The word “DARPin” is an acronym for “Designed Ankyrin
Repeat Protein.” An ankyrin repeat is a motif that
consists of two helices separated by a loop, as shown
below. The number of helix-loop-helix repeats varies
from four to thirty-four.

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The ankyrin repeat motif is only one of a number of runaway growth. Molecular Partners is developing
secondary protein structures, but for our purposes, Abicipar in partnership with Allergan (Dublin, Ireland).
it’s the only one that matters. It mediates a wide Molecular Partners also has two clinical candidates in
variety of protein-protein interactions. The number of oncology. The first, MP0250 is in Phase II development
repeats present influences the final shape of the motif, for multiple myeloma and non-small cell lung cancer.
and so determines what target protein the ankyrin MP0250 is a “Multi-DARPin,” which means that it
repeats interact. can recognize two targets at once. This strategy has
important implications. Tumors mutate very rapidly,
so they often develop resistance to therapies. By
targeting two cancer-associated proteins simultaneously,
researchers hope to overcome this challenge. MP0250
binds to and inhibits both VEGF and hepatocyte
growth factor (HGF). Tumors secrete VEGF to promotes
angiogenesis, or the growth of blood vessels that supply
nutrients into a tumor. Inhibiting VEGF can help starve
the cancer. HGF promotes growth of the tumor cell itself.
The second oncology drug candidate, MP0274, is aimed
at HER2-positive breast cancer, a form of the disease
that produces extra copies of the HER2 growth-factor
SIMPLER, CHEAPER, FASTER
receptor protein. MP0274 binds to two different sites
PROTEIN POWER
on HER2 and induces apoptosis (cell suicide). This
The founders of Molecular Partners (Zurich, mechanism differs from that of the mAb Herceptin.
Switzerland) genetically engineered a large library of Herceptin also targets HER2-positive breast cancer.
ankyrin repeat proteins—the aforementioned “DARPins.” However, it binds to a different HER2 site and blocks
These DARPINS are then screened for their ability to growth factor signaling. It also activates the patient’s
interact with and modulate a variety of drug targets, with immune system to attack the tumor. MP0274 is in Phase
the same high specificity of mAbs. 1 clinical testing for HER2-positive cancers, including
DARPins’ structure is far less complex than that of mAbs. breast and gastric cancers.
Consequently, companies can use bacteria cells to
manufacture them—which translates into much cheaper PRECLINICAL DEVELOPMENT
production costs. The relative “simplicity” of DARPins In partnership with Amgen (Thousand Oaks, CA),
may also mean that researchers will have an easier time Molecular Therapeutics also has a DARPin entering the
formulating them for therapeutic use. And yes, you immuno-oncology world. MP0310 is designed to activate
guessed it, DARPins are also small in relation to mAbs. immune cells only in a specific tumor, not throughout a
Their size may enable them to penetrate hard-to-reach patient’s body. Now in preclinical development, MP0310
targets like solid tumors. These characteristics mean features two domains. The first domain recognizes
that at least for some uses, DARPins may prove more and binds a tumor-specific protein. The other binds
effective than their bigger biologic cousins. CD40, a protein on the surface of white blood cells that
activates them. The white blood cells will only activate
EYES ON DARPINS if the tumor-specific domain is also engaged. This fail-
Molecular Partners’ most advanced DARPin therapeutic, safe restricts the DARPin’s activity to tumor cells, which
Abicipar, is in Phase III clinical development for wet could limit side effects that occur with treatments that
age-related macular degeneration (AMD), a serious eye systemically activate the immune system.
condition. Wet AMD is caused by excessive growth in the DARPINs are still in development. Only time will tell if
arteries and capillaries that supply the retina with blood. they will become as effective and versatile as mAbs. We
The overgrowth leads to retinal detachment, which can look forward to following the story about what just might
ultimately cause blindness. Abicipar works by binding be one of the next big things in biotech.
to and inhibiting the growth factor, vascular endothelial
cell growth factor (VEGF) that triggers the blood vessels’

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$8.7 BILLION SMA DRUG EXPLAINED Life Science Training from Industry Experts

THE SCIENCE BEHIND THE DEAL An orphan disease, SMA affects about 1 in 10,000 babies
born in the United States. The four generally accepted
Earlier this week, news of the $8.7 billion acquisition
classifications of SMA are:
of gene therapy company AveXis (Bannockburn, IL)
by Novartis (Basel, Switzerland) made big biotech • Type 1: The most severe and the most common type
headlines. AveXis’ lead candidate, AVXS-101, is now of SMA. Symptoms are usually present within the
in Phase III clinical studies for the treatment of first few months of life, and these babies often do
spinal muscular atrophy (SMA). In this Weekly, we’ll not display movement of any kind. As the disease
take a look at the science behind these headlines progresses, toddlers have trouble with swallowing
by explaining exactly what SMA is and describe the and respiratory function. SMA Type 1 is usually fatal
different therapeutic approaches being used to tackle by age two.
this disease. • Type 2: Symptoms manifest between six and
eighteen months. These children can typically sit
SMA PRIMER but not stand or walk. Respiratory function is often
Our nervous system consists of the brain, spinal cord, compromised and is a major concern, however with
and a vast network of nerves that feed into every tissue the help of machines many of these patients live
of the body. Motor neurons are a type of nerve cell into adulthood.
that sends messages from the spinal cord to muscles, • Type 3: Symptoms occur after age one. These
enabling movement. patients are usually able to walk, but may lose
In order for the motor neurons to do their job, a that ability as the disease progresses. Respiratory
functional protein called the survival motor neuron function is less impaired, and life expectancy is often
(SMN) protein is necessary. The survival motor neuron near normal.
1 (SMN1) gene is responsible for producing most of the • Type 4: This is the adult-onset form, typically
SMN protein used by the body. A second, closely related manifesting at age 30 or later. Muscles gradually
gene is the survival motor neuron 2 (SMN2) gene, which weaken, and the patient often needs to use
produces a much smaller amount of SMN protein and is a wheelchair later in life. Life expectancy is
seen as a sort of “back-up” version to SMN1. not affected.
SMA is caused by a variety of mutations in the SMN1 Because SMA Type 1 is the most common and severe –
gene. Without functional SMN protein, the neurons do about 60% of cases – most of the drugs in development
not work correctly and eventually die. How soon they aim to tackle this portion of the disease population.
die depends on the extent of the SMN deficiency, which Successful therapies will likely later be tested in the less
correlates with the severity of the disease: the less SMN severe forms of SMA. Below we list some approaches
produced, the more severe the disease. going after the root cause of this orphan disease—not
The back-up gene, SMN2, produces a small amount of enough SMN1 protein production.
functional SMN protein. However, differences in the way
SMN2 functions means most (but not all) of the protein MAKING SENSE FROM ANTISENSE
is non-functional and degrades shortly after being Spinraza (Biogen; Cambridge, MA) is the only drug
produced. Patients with less severe forms of the disease currently on the market to treat SMA. As an antisense
usually have extra SMN2 copies because ultimately, drug, Spinraza is a short, synthetic piece of RNA whose
even tiny amounts of SMN protein provides some motor sequence directs it to bind to the SMN2 mRNA (recall
nerve function. that mRNA is produced from a gene, and contains the
information used by the cell to produce the protein
specified for by the gene). The binding of Spinraza

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changes the way cells process the SMN2 mRNA, causing As a one-shot treatment, AVXS-101, if approved,
more of the information in the mRNA to be converted offers the potential of a one-and-done cure for SMA,
into protein. The end result is a greater amount of full- differentiating it from Spinraza’s three-times-a-year
length, functional SMN protein. dosing schedule.

GENE THERAPY CURE? SMALL MOLECULE ENHANCERS

As a single gene disorder, SMA is an ideal candidate for PTC Therapeutics (South Plainfield, NJ) — in partnership
gene therapy approaches because delivering a “good” with Roche (Basel, Switzerland) — has begun Phase
copy of the mutated gene could potentially cure the II clinical studies on its proprietary small molecule
disease by supplying a permanent copy of the correct drug, RG67916. RG67916 is similar to Spinraza in that it
SMN1 protein-making instructions. changes the way nerve cells process the SMN2 mRNA,
Using a “vector” — a virus stripped of its disease-causing resulting in increased production of functional SMN
ability — scientists are able to safely deliver corrected protein. However, there is a notable difference is the
genes into targeted cells. In the case of SMA type 1, the delivery mechanism — Spinraza is an injectable while
AAV9 vector crosses the blood-brain barrier and delivers RG67800 is a pill.
corrected copies of the SMN1 gene into motor neuron SMA is a truly devastating disease. Since its genetic
cells in the brain. basis was first discovered two decades ago, researchers
AveXis’ AVXS-101 has generated excitement in large part have made substantial progress in understanding the
due to clinical trial observations of babies who received disease and coming up with ways to target its molecular
this gene therapy showed marked increases in SMN basis. 2018 might just be the year when that molecular
production and in movement – with most participant understanding translates into the closest thing yet to
even talking and sitting without assistance. If additional a cure.
studies confirm these results, the breakthrough-
designated drug could be filed for FDA-approval later
this year.

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PAPA’S GOT A BRAND NEW LAB COAT Life Science Training from Industry Experts

PAPA’S GOT A BRAND NEW LAB COAT JUST ADD WATER

Last month, Johnson & Johnson (New Brunswick, NJ) This biosensor is embedded in an everyday substance:
announced the winners of its “Lab Coat of the Future” paper. Not available at your local Staples, this stuff is
competition. This contest challenged participants to known as paper-based synthetic gene networks. The Lab
combine new technology and good design to transform Coat of the Future competition meant figuring out how
the age-old symbol of science and medicine—the white to make the reaction work with sensors embedded in a
lab coat—into a symbol of breakthrough innovation. different medium—fabric.
The winners, a group of researchers from Harvard To produce a particular protein, a gene needs a stew of
University’s Wyss Institute who call themselves enzymes and amino acids, the building blocks of new
TeamBioFabric, developed a lab coat prototype that proteins. Outside the lab, this process happens within
detects whether its wearer has been exposed to a cell. For the past few decades, researchers have
hazardous substances, including pathogens and toxins. replicated the process in test tubes by creating a “soup”
The prototype builds on the group’s success developing of the necessary ingredients. The test tube environment
synthetic biology biosensors—DNA-based systems works well for basic research; for developing portable
that can detect, analyze and alert users to an external biosensors, not so much. The Wyss scientists found that
biological threat. Let’s take a closer look. they could make a biotech Soup Starter—freeze-drying
the ingredients—and embed it in small paper discs.
CANARIES IN THE COAL MINE The Soup Starter discs remain inactive and stable for
Biosensors have been around a long time. Think months. Rehydrating a disc resuscitates the biosensor.
about the little birds that for much of the 20th century If a drop of blood, for example, containing the trigger
alerted miners to invisible, odorless and deadly carbon molecule hits the disc, a color change occurs. With Zika,
monoxide. Electronic sensors replaced the feathered the color morphs from yellow into purple.
yellow sentinels in 1986. This millennium takes
biosensors to the molecular level. POWERFUL DIAGNOSTICS:
OUTSIDE AND INSIDE THE LAB
Like Tweety and his cousins, synthetic biology
biosensors detect an input, MRSA (methicillin-resistant These “freeze-dryable” biosensors may have a
Staphylococcus aureus) for example, and produce an significant impact in the field of public health. The low-
output that warns of danger. For the poor little canaries, cost, portable on-site diagnostics can quickly detect
the output was death. For wearers of the Wyss Institute the presence of deadly viruses such as Ebola or Zika.
“Super Lab Coat,” the output is usually a color change or Early diagnosis could help contain outbreaks, saving
fluorescent glow on the part of the coat enhanced with countless lives.
the biosensor. These simple, powerful little papers inspired Team
In 2016, the Wyss Institute team developed a biosensor BioFabric’s entry into the Lab Coat of the Future contest.
that detects Zika virus, designated as a burgeoning Instead of paper, the researchers wove the freeze-dried
global health emergency by the World Health biosensors into fabric. The idea is to warn health-care
Organization. The Wyss biosensor used a gene sequence providers as soon as possible of exposure in the lab or
that, in the presence of Zika virus genetic material, doctor’s office to an infectious agent. The biosensors can
is triggered to make a protein. The new protein, an be easily adapted to a range of pathogens.
enzyme, catalyzes a reaction that manifests in a color
change—thus indicating the presence of Zika.

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SUPER LAB COAT 2.0 exposure alert. Busy doctors or researchers may not
notice a color change, but just try to ignore a cellphone!
With their $50,000 prize money, Team BioFabric is kicking
their super lab coat up another notch. The new goal is The super lab coat and its sibling, SLC 2.0, may not make
to integrate the freeze-dried biosensor with wearable New York Fashion Week in September, but that’s OK by
electronics and embedding both into a lab coat. In us at Biotech Primer WEEKLY. We’re putting our money
addition to changing color, the even more souped-up on the new, intelligent lab coats anyway.
lab coat would ping the wearer’s cell phone with an

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TICKED OFF: THE SCIENCE
BEHIND LYME DISEASE Life Science Training from Industry Experts

TICKED OFF: THE SCIENCE THE INFAMOUS BULLS-EYE

BEHIND LYME DISEASE The most notorious sign of Lyme disease is the red bulls-
Alleluia! Spring! It means the promise of more time in eye rash that often develops near the bite. According to
the fresh air – gardening, camping, or just enjoying the the Center for Disease Control, other symptoms that
sunshine. Unfortunately, time outdoors increases the may occur within the first 30 days of infection include
risk of contracting Lyme disease. As you may know, this fever, muscle and joint aches, and swollen lymph nodes.
illness comes courtesy of our favorite blood-sucking Unfortunately, not everyone develops the bulls-eye, and
arthropods: ticks. Not just any tick, however. Only the it’s easy to confuse Lyme disease with other illnesses.
diminutive deer tick (Ixodes scapularis) transmits this Failing prompt diagnosis and antibiotic treatment, a
nasty disease. Lyme has been reported in every US state plethora of more severe symptoms can develop days or
but most commonly occurs in the Northeast and the even months after exposure. These can include heart
upper Midwest. The disease gets its name from the tiny palpitations, severe joint pain, facial palsy, and short-
Connecticut town of Lyme, where doctors identified a term memory issues.
cluster of cases in the 1970s. This WEEKLY looks at the
cause, treatment, and possible complications of this
potentially debilitating disease. We’ll also preview a new
Lyme vaccine that the FDA has awarded fast-track status.

TRICKY DIAGNOSIS
Health care providers primarily diagnose Lyme disease
based on symptoms, known or potential exposure to
deer ticks, and an FDA-approved blood test for anti- B.
OH DEER burgdorferi antibodies. None of these approaches is
The microbiological culprit behind Lyme is the bacterium ideal. Lyme disease’s rash fails to appear in about a
Borrelia burgdorferi. These microorganisms inhabit deer quarter of infected people. Unfortunately, it usually
of course, but smaller animals such as mice and birds takes a month or longer after infection for people to
too. A hungry deer tick chomps on, say, an infected produce enough B. burgdorferiantibodies to show up in
white-footed mouse. When the tiny black-legged parasite a blood test. This lag can delay treatment and creates
eventually feeds again, it passes on the gift of Lyme to its a window for the most serious outcomes such as nerve
next victim, whether gardener or groundhog. damage and heart problems. Thus, epidemiologists and
In a human host, proteins on the surface of B. burgdorferi other scientists are in hot pursuit of tests capable of
wreak havoc within and outside our cells. The bacterium detecting the bacterium earlier.
disrupts the function of infected tissues at the cellular
level. Combined with the body’s inflammatory response,
THE NANOTRAP
the symptoms of Lyme disease ensue. One innovative approach comes from Ceres
Nanoscience (Manassas, VA) and their “nanotrap”

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technology. Rather than trying to detect a patient’s TREATMENT
response to infection, the Ceres Nanotrap Lyme Antigen
People whose Lyme disease gets diagnosed early
Test detects the presence of B. burgdorferi itself. It
are most likely to respond to the standard antibiotic
works by identifying the bacterial protein OspA in the
treatment. Otherwise, debilitating symptoms are more
urine of infected patients. Other researchers have tried
likely to develop and linger. In a small percentage of
this method but found themselves stymied due to the
patients, symptoms such as fatigue and pain linger for
low concentration of the protein in the samples. Enter
months or even years. This phenomenon is known as
nanotrap technology. These nanoparticles (10-9 meter
post-treatment Lyme disease syndrome.
diameter) consist of an outer shell that surrounds a
“bait” molecule. The shell filters out proteins larger than
the target. Meanwhile, the molecular bait inside captures
COVER UP & SPRAY!
only OspA. The nanotraps are then processed to remove For now, the best way to avoid contracting Lyme disease
any captured proteins and identify them. is to protect yourself by wearing long sleeves, pants, and
a hat or scarf in areas where deer ticks are common. In
The new Ceres test is available as a CLIA (Clinical
addition, it’s important to check yourself or your loved
Laboratory Improvements Amendment) test. This means
ones for ticks after you’ve spent time outdoors.
that the test itself cannot be sold to a physician’s office
or pharmacy but must be performed at a laboratory
VACCINE ON THE HORIZON
facility approved by the Centers for Medicare and
Medicaid services. In suspected Lyme disease cases, Fortunately, a French company, Valneva, is giving lovers
a urine sample is sent to the Ceres lab, analyzed using of the great outdoors something more than a bottle of
the Nanotrap test, and the results sent back to the DEET to ward off Lyme disease. The Lyon, France-based
physician’s office. biotech company is developing a Lyme vaccine. Known
as VLA15-101, it consists of the B. burgdorferi surface
PCR-BASED TESTING protein OspA. Researchers hope to train our bodies via
needle to recognize the bacterium as a threat and rev
Another strategy for diagnosing infectious diseases
up our immune response in case of actual infection. The
sooner rather than later is PCR-based testing -- detecting
vaccine includes all six types of OspA proteins. VLA15-
and amplifying specific pathogen-associated gene
101 is currently in Phase I clinical studies and has been
sequences. This approach works well with many viruses,
granted fast-track status by the FDA.
including HIV and influenza. However, identifying a
bacterial infection such as Lyme disease this way is
COCKTAIL FODDER: FORMIDABLE FOES
tougher. Why? During an active viral infection, viruses
reach high concentrations within the bloodstream. This Ticks have a few tricks up their teeny-tiny segmented
makes it relatively easy to find them in a simple blood sleeve when it comes to obtaining their blood banquet.
draw. In contrast, the Lyme bacterium tends to “hide People don’t always notice a bite because the sneaky
out” in tissues, making it more difficult to obtain gene little arachnids inject a neurotoxin (basically a local
sequences from a blood sample. Nonetheless, there is anesthetic) when they latch on. By the time the host
a DNA-based test for the disease, developed by DNA finds the freeloader, the little parasite can be very
Connexions (Colorado Springs, CO). Like the nanotrap difficult to remove. That’s because the tick secretes
test, it’s available as a CLIA lab test. another secret weapon – a sticky substance called
“cementum” near the bite, allowing it to hang on tight for
chow time.

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IMMUNOTHERAPY GOES VIRAL Life Science Training from Industry Experts

VIRUSES BLASTING CANCER simplex 1 virus. The modifications made to Imlygic to

ensure safety and efficacy include:
Last week, Johnson & Johnson (New Brunswick, NJ )
announced that their biotech arm Janssen Labs (Rariton, • Deletion of viral gene ICP34.5, enabling Imlygic
NJ) is investing up to $1 billion dollars to acquire BeneVir, to replicate only in cancer cells (not in healthy
a small biotech based in Rockville, Maryland, which human cells.)
focuses on oncolytic virus development. Earlier this year, • Deletion of viral gene ICP47, helping the virus evade
Merck (Kenilworth, NJ) acquired Australian oncolytic immune detection.
virus company Viralytics (Sydney). Once considered a • Increased activation of the viral gene US11, resulting
long shot, this type of immunotherapy is now being in more viral replication in tumor cells so a larger
taken seriously by bluechip biopharma players. In number of cancerous cells are killed.
this WEEKLY, we’ll review the fundamentals of how
• Insertion of a gene for the human protein GM-CSF,
oncolytic viruses work and take a look at the increasingly
which “revs up” the immune system, aiding in the
crowded pipeline.
overall immune response against the tumor.
ONCOLYTIC VIRUS PRIMER These modifications create a virus that selectively
Oncolytic viruses are an immunotherapy—a type of replicates in tumor cells, resulting in their direct
therapy that harnesses the power of a patient’s immune destruction as well as activation of a host immune
system to combat a disease. Getting a virus to trigger response targeting the virus-infected tumor cells.
an immune response to fight cancer is no easy task.
Oncolytic viruses are created in the lab by genetically
modifying existing viruses in at least two ways:
1. Making the virus safe by removing genes that cause
sickness in people
2. Engineering the virus to recognize and kill cancerous
cells and disregard healthy, non-cancerous cells
The oncolytic virus follows the same life cycle as any
virus—once inside the human body it hunts down and
enters its host cell. In this case, the host happens to be
cancer cells. As the oncolytic virus multiplies inside of the
tumor, it causes these cells to burst open, killing them. STEALTHY INVADERS
Spewing from the burst are new virus particles that BeneVir’s oncolytic virus caught Janssen Lab’s attention
target remaining tumor cells. The virus also activates the because it incorporates the company’s “T-stealth”
immune system, bringing in a second line of attack.. technology, an oncolytic virus platform also built on
Most oncolytic viruses are being tested as both stand- a herpes simplex 1 virus that selectively infects and
alone treatments and in-combination with other kills tumor cells. The T-stealth platform incorporates
immunotherapies such as checkpoint inhibitors. an additional viral gene that prevents T-cells from
recognizing and attacking the therapeutic virus itself.
INSIDE IMLYGIC This innovation should allow the virus to spread to more
cells within a tumor. The T-stealth oncolytic viruses
Amgen’s (Thousand Oaks, CA) Imlygic, which targets
are in preclinical development for the treatment of
melanoma, is currently the only FDA approved oncolytic
solid tumors.
virus. The virus used in Imlygic is a modified herpes

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ONCORUS ON THE OFFENSE oncolytic virus to penetrate the tumor and deliver genes
instructing the tumor itself to make the antibody could
Another modified herpes virus, ONCR-001, is being
be a game-changing work around.
developed by Oncorus (Cambridge, MA) for the
notoriously difficult-to-treat brain cancer, glioblastoma. ONCOLYTIC PIPELINE
Like Imlygic, ONCR-001 has been modified to selectively
target tumor cells. Instead of blocking viral replication,
Oncorus scientists engineered a “suicide switch” into
their virus that is activated only in healthy cells. When
ONCR-001 infects healthy cells, the switch is triggered,
and the virus is destroyed. The switch is not triggered by
tumor cells, leaving the virus fully able to kill the enemy.

THE GENESIS OF GENELUX

San Diego-based Genelux is adapting the vaccinia virus
as an oncolytic virus for the treatment of a variety of
solid tumors. Vaccinia is already used as a vaccine for
smallpox, meaning that it already has a decades-long COCKTAIL FODDER: RENEGADE
safety record, although the modified version must still VIRUSES OF THE PAST
undergo safety testing.
The idea of using viruses to challenge cancer is
Their lead product, GL-ONC1, selectively replicates in cutting-edge, 21st-century science, but the inkling of
tumor cells and tumor-associated blood vessels, directly a cancer-fighting virus was first observed more than
killing tumors while cutting off their blood supply. a century ago. In 1904, an editorial published in the
GL-ONC1 is in Phase II clinical testing for a variety of American Journal of Medical Science revealed a
solid tumors. spontaneous regression of cervical cancer occurred
The company is also developing oncolytic viruses after administration of a rabies vaccination. A few years
with genes for “therapeutic payloads” — proteins and later, a similar phenomenon occurred: the remission
therapeutic antibodies that will boost the patient’s of lymphoma after a measles virus infection. Our
immune response to the cancer because these payloads modern understanding of viruses at the molecular level
will be produced inside the cancer cells. This approach combined with our increased ability to manipulate genes
is a clever response to the fact that due to their made this century-old idea a medical reality of today.
relatively large size, most therapeutic antibodies are
not able to completely penetrate solid tumors. Using an

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TAKING A SWING AT PEANUT ALLERGIES Life Science Training from Industry Experts

HOW DO ALLERGIES DEVELOP? TERM OF THE WEEK: HISTAMINES

Every summer, watching a game at the ballpark Histamines are small molecules produced by mast cells.
and digging into a bag of peanuts is a source of Once released, histamines bind to receptors on the
entertainment for many Americans. For the 15 million surface of blood vessels, increasing their permeability;
who suffer from peanut allergies, the idea of being taken the ease with which fluid moves out of the blood vessels.
out to the ballgame elicits concern — or even anxiety. Histamines also bind to receptors on certain types of
Food allergies — think tree nuts, milk, eggs, wheat, nerve cells, resulting in muscle contraction. A host of
soy, fish, and shellfish — are on the rise. The mere dust symptoms can be triggered, ranging from annoying to
particle of a freshly cracked peanut can be responsible deadly. Typical signs of histamine release include:
for an unpredictable cascade of reactions, including • Increased blood vessel permeability resulting in a
death brought about by anaphylaxis. runny nose and watery eyes.
This WEEKLY takes a swing at explaining how allergies • Increased muscle contraction leading to throat
develop, the current treatments, and new products that constriction, difficulty breathing and swallowing.
might change the way allergen desensitization therapy • Extreme fluid release from tissues causing a sudden
is delivered. drop in blood pressure, potentially bringing on a
heart attack.
SOMETHING TO SNEEZE AT
Mild allergy symptoms such as runny nose and watery
The host of symptoms dubbed “allergies” are the
eyes can often be successfully controlled by the use of
end result of the immune system’s response to a
an over-the-counter antihistamine, a drug that works by
normally harmless substance, as if that harmless
blocking the interaction of histamines with receptors on
substance were a threat. An initial allergen exposure
nerve and muscle cells. However, once anaphylaxis, a
results in the production of a class of antibodies called
severe allergic reaction that includes difficulty breathing
Immunoglobulin E (IgE). A second exposure to the
and heart palpitations, has occurred, it is too late for
allergen results in an “allergen-IgE antibody complex.”
antihistamines to be effective. These symptoms can only
These newly-produced complexes bind to and activate
be treated with an injection of the hormone epinephrine.
mast cells—a type of immune cell. As the image below
And the sooner the injection the better in cases where a
shows, activated mast cells send out chemical alarms in
life is on the line.
the form of histamine.
Epinephrine helps to reverse histamine’s effects by
decreasing blood vessel permeability, relaxing muscle
cells, and stimulating the heart. People at risk for
anaphylaxis need access to an epinephrine auto-injector,
a spring-loaded syringe that makes the lifesaving shot
readily available. This type of product is referred to as a
“combination product” because it combines a device (the
auto-injector) with a medicine (epinephrine). Mylan’s
(Canonsburg, PA) EpiPen is an epinephrine auto-injector,
as is Amedra’s (Horsham, PA) Adrenaclick.

THE HYGIENE HYPOTHESIS

Epidemiologists have noticed an interesting trend as
countries transition from developing to developed
status: an improvement in sanitation and access to

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antibiotics means less pathogenic exposure and lower ALLERGENS BY THE DOSE
infection rate. As the infection rate drops, the incidence
The current allergen immunotherapy market includes
of allergies shoots up.
allergy shots (which require monitoring by a physician)
Many scientists think early exposure to infection helps and drops or tablets dissolved under the tongue
shift the immune response towards fighting pathogens (which can sometimes be taken at home). Aimmune
while minimizing the production of IgE antibodies. Therapeutics’ (Brisbane, CA) AR101 is a capsule
Exposure to potential allergens (while the immune containing pharmaceutical grade peanut protein. The
system is still developing) helps to desensitize the patient opens up the capsule and mixes the contents
allergic response. with food as a means of delivery.

THE STRATEGY BEHIND AR101 clinical studies report patients becoming

DESENSITIZATION THERAPY desensitized to doses at least twenty times greater
than the original allergy-inducing dose and in some
The idea of allergy desensitization through controlled
cases, more than 100-fold greater. The goal is to
exposure has been around for decades. Desensitization
reach a tolerance level that offers protection against
is the principle behind allergy shots shown to be
accidental eating of peanuts. The product was awarded
effective against pet dander, dust mites, and pollen.
a Breakthrough Designation by the FDA and has recently
Desensitization therapy was once considered to be too
completed Phase III. Aimmune Therapeutics plans to file
risky for food allergies, but a number of new studies
for FDA approval by the end of 2018.
support the idea that gradual exposure to food allergens
may be beneficial. DBV Technologies (Bagneux, France) is also in the
allergy fight. Their Viaskin skin patch delivers low
The latest National Institutes of Health (NIH)
doses of either peanut, milk, or dust mite allergens. By
guidelines recommend the early introduction of peanuts
delivering an allergen through the skin instead of the
into children’s diets, including those considered to be
blood, the body will react less severely, reducing the
at high risk for developing peanut allergies because
risk of anaphylaxis. The Viaskin peanut patch has been
they already have severe eczema or egg allergies.
awarded Breakthrough status and has completed Phase
These new guidelines are based on the results of the
III clinical studies. DBV is expected to file a Biologics
NIH-funded Learning Early About Peanut Allergy (LEAP)
Licensing Application with the FDA in the second half
study, which randomly assigned 600 high-risk infants to
of 2018.
either peanut-avoidance or the regular inclusion of small
amounts of peanut products in their diet for the first As allergen desensitization treatments continue to make
five years of life. At age five, peanut allergy was assessed their way through the drug pipeline, allergy sufferers
and an 81 percent reduction in allergy was found in remain hopeful for better and easier treatment options,
the children who regularly consumed peanuts when and, maybe even one day, a cure.
compared to those who avoided them.

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CRISPR TO THE PEOPLE Life Science Training from Industry Experts

CRISPR: SNIPPING AWAY AT DISEASE • The bacteria make the DNA-cutting enzyme Cas,
which attaches to the new viral RNA.
CRISPR genome editing is one of the most exciting
developments in biotechnology since its discovery a • The resulting viral RNA/Cas complex finds its match
few years ago. Bacteria use this mechanism to destroy on the invading viral DNA.
the DNA of invading viruses. Scientists subsequently • The RNA attaches to the DNA and the Cas enzyme
discovered CRISPR’s potential for making therapeutic cuts up the foreign DNA, destroying the virus.
changes to the human genome. CRISPR therapies • Voila—“healthy” bacteria.
entered clinical trials last year. Now this amazing
technology is evolving into second-generation CRISPR TO THE PEOPLE!
applications such as RNA-editing and CRISPR-based
In 2013, researchers adapted this defense for use in
diagnostics. This WEEKLY reviews how the genome
human cells. By adding a “guide RNA” and Cas enzyme to
editor works and explores new applications. Next week,
target a specific DNA sequence, scientists demonstrated
we preview how researchers are adapting CRISPR to
the system could be used to cut human DNA in precise
diagnose disease.
locations! This original Cas protein came from the Cas9
Streptococcus bacteria—hence the moniker CRISPR/
BAC FIGHTS BACK
Cas9.
CRISPR is a key immune response in bacteria. Yes,
What makes CRISPR/Cas9’s ability to cut human DNA in
Virginia, bacteria have teeny tiny little immune
precise locations so cool? The protein creates double-
systems. Like we do, these microorganisms fall prey
stranded breaks (DSB) in the specified DNA sequence.
to viral infection. (Take that, salmonella!) They’ve
Double-stranded breaks cut both strands of the
evolved a fascinating way to repel the invaders. In
DNA helix.
the 1980s, scientists observed a pattern in bacterial
genomes: Repeating, palindromic sequences, with Think of DNA as a two-lane bridge. Now imagine an
unique sequences—“spacers”—between repeats. earthquake takes place causing one section to break off,
They bestowed a tongue twister of a name, “clustered falling away. DSBs can repair the damaged DNA bridge in
regularly interspaced short palindromic repeats,” on the two ways:
mechanism, which we happily call CRISPR. Scientists also • Homology Directed Repair (HDR) relies on a highly
noticed CRISPR sequences always occur near genes that similar DNA segment to repair the break. In this case,
code for an enzyme that cuts DNA. This enzyme became workers build a new section of the genetic bridge
known as Cas, or “CRISPR-associated.” offsite and then helicopter it into place.
In the mid-2000s, scientists realized these spacers • Non-Homologous End-Joining (NHEJ) closes
matched the DNA sequences of infecting viruses. The the gap using another strategy. Visualize workers
sick bacteria were stashing bits of the offending viral pushing the two remaining sections of the bridge
DNA between its own CRISPR sequences! These viral back together. NHEJ can result in a sequence error,
DNA snippets create a “genetic memory,” which enables just as sections of a repaired bridge often don’t line
the bacteria to fight back if reinfected. Here’s how: up properly. If the repair occurs in the middle of a
• Viral DNA present in the spacer sequences is copied gene, it typically disrupts gene function and halts the
into viral RNA. production of the matching protein.

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 RCas9 cuts mRNA. Scientists can target specific disease-
associated mRNAs with RCas9. The UCSD team has
tested RCas9 on cell-based disease models in the lab
and have shown RCas9 to reduce problematic mRNAs
in Huntington’s disease, myotonic dystrophy, and
amyotrophic lateral sclerosis models.
Targeting disease-associated RNAs instead of genes may
allow researchers to treat certain genetic diseases by
getting at their root cause without permanently altering
a patient’s genome. The UCSD team has launched
Locana (San Diego, CA) to move this technology from the
By engineering double-stranded breaks at specific lab into the clinic.
locations, scientists trigger the NHEJ or HDR cell
repair pathways.
THE LATEST MODEL OF
GENETIC SCISSORS: CAS13
• Activating NHEJ disrupts a disease-associated
Cas9 is only one enzyme used in CRISPR . Its utility
gene. This prevents the production of the protein
has inspired scientists to hunt for related proteins
that causes the disease.
with slightly different applications. One of the
• Activating HDR fixes mutated genes by most interesting is the Cas13 enzyme. Like RCas9, it
simultaneously delivering a “repair template” that targets mRNA.
contains the correct gene sequence.
Scientists at the Broad Institute in Cambridge, MA,
Both scenarios present possible cures for different types modified Cas13 to edit single bases that comprise
of disease. For example, CRISPR is currently in clinical the mRNA sequence instead of just cutting mRNA.
trials at Sichuan University (Sichuan, China). Here, Specifically, the new and improved Cas13 changes the
researchers are delivering CRISPR/Cas9 components base adenine (A) to inosine (I). This inosine base isn’t
to cancer patients’ white blood cells to disable the normally found in mRNA, but our cellular enzymes
PD-1 gene. The PD-1 gene inhibits these immune cells. recognize it as the common base guanine (G). G to A
By deactivating the PD-1 gene the immune system mutations play a role in Duchenne muscular dystrophy
is left intact and the patient should be able to more and Parkinson’s disease.
aggressively fight cancer. Next on the research agenda
Repairing mutated mRNA may restore a patient’s
come trials aimed at correcting gene sequences mutated
normal protein function without permanently changing
in sickle cell disease and hereditary blindness.
their genome. The Broad Institute team christened
RCas9—THE RNA-EDITOR their upgraded Cas13 enzyme REPAIR, RNA Editing for
Researchers at the University of California, San Diego Programmable A to I Replacement.
(UCSD), have developed RCas9, a modified version The ability to edit RNA significantly expands CRISPR’s
of Cas9. RCas9 targets messenger RNA (mRNA), the therapeutic usefulness. Equally important, it eases
intermediary between DNA and proteins. Information potential safety concerns that could arise from making
stored in a gene (DNA) is converted into mRNA, which lasting changes to someone’s DNA.
is then used by cells to make a protein. In other words,
Next week, we look at the flip side of therapeutic
mRNA is a temporary copy of the permanent genetic
CRISPR—its use in diagnostics.
information DNA stores.

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CRISPR’S GENOME DETECTIVES Life Science Training from Industry Experts

CRISPR’S GENOME DETECTIVES The test has been adapted for use as a “lateral flow
assay” –similar to an at-home pregnancy test. Think First
Last week, we reviewed how CRISPR works and its
Response for Zika.
potential to revolutionize genetic therapies. Here we look
at how scientists have started using this technology to
develop new diagnostics.
Media attention has been focusing almost exclusively on
how scientists use CRISPR to edit DNA. But as you read
last week, the biotech industry has begun to turn its
attention to new bacterial enzymes that work with guide
RNAs to target specific regions of the genome.
One enzyme, Cas13, has another feature that enables
it to work as a diagnostic tool as well as a therapeutic
one. No doubt you remember from last week that
scientists first inactivate Cas13 and then alter it to edit DETECTR
a single base. But this nifty little enzyme is a twofer!
UC Berkeley scientists have developed a similar tool:
Cas13 chops up not just its target mRNA, but any other
DETECTR. This diagnostic, DNA Endonuclease Targeted
nearby mRNA too. Researchers call this mass destruction
CRISPR TransReporter, works with another Cas enzyme,
“collateral cleavage.”
CAS12a. This enzyme cuts a specific DNA sequence with
the help of guide RNA. Like Cas13, it then goes nuclear,
SHERLOCK: THE RNA GAME IS AFOOT!
attacking nearby DNA sequences. In this case, the
The Broad Institute (Cambridge, MA) has developed a destruction includes fluorescently-labeled DNA reporter
Cas13-based diagnostic consisting of a Cas13/guide RNA segments. The Berkeley team has demonstrated
combo that targets virus-associated sequences. Some of DETECTR’s ability to identify the human papillomavirus
the world’s most dangerous viruses, including Zika and (HPV) in a patient blood sample. DETECTR should also
HIV, have RNA-based genomes. be able to detect cancer and other disease-associated
If a sample contains a targeted sequence, Cas13 homes mutations in a patient DNA sample. Last month, San
in on it and cuts it. Easy peasy, right? But how do Francisco-based Mammoth Biosciences was launched
diagnosticians know if the enzyme has done its job? to commercialize the development of this exciting new
Short answer: reporter RNA comes with a label that diagnostic platform.
can glow! CRISPR-based diagnostics promise extreme sensitivity
A label? No, not paper. Molecular—the reporter RNA – both the ability to detect vanishingly small amounts
releases and activates a fluorescent tag only if and of DNA or RNA–and specificity–detection based on a
when the viral sequence has been cut. That is-- only if specific gene sequence. This potent combination of
the Cas13 enzyme finds its target and is activated. The attributes lays the groundwork for finding infectious
Broad team calls the new diagnostic “Specific High- diseases and cancers faster.
sensitivity Enzymatic Reporter unLOCKing” – SHERLOCK.

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GROUNDBREAKING MIGRAINE DRUG EXPLAINED Life Science Training from Industry Experts

FIRST IN CLASS MIGRAINE APPROVAL • Brain Stem Changes: Research by the Mayo
Clinic (Rochester, MN) suggests migraines derive
Last month the FDA approved Amgen’s (Thousand Oaks,
from changes in the brain stem and its interaction
CA) new migraine drug Aimovig, the first drug shown to
with the trigeminal nerve. The trigeminal nerve
prevent the onset of migraines. The drug significantly
supplies feeling to the face and is considered a pain-
reduces the number of migraine days in difficult-to-
associated pathway in migraine attacks.
treat (those that have failed 2 to 4 prior treatments)
patient populations. In some patients dubbed “super • Lowered Serotonin Levels: Another area of active
responders”, migraines occurrence went from several research involves the neurotransmitter serotonin.
times/month to no occurrence for 6 months. This is a Serotonin is most often associated with mood—
big breakthrough that will have significant impact on the antidepressants such as Eli Lilly’s (New York, NY)
quality of life for migraine suffers who have not been Prozac, increases levels of serotonin in the brain.
able to find other forms of relief. This neurotransmitter is also implicated in migraine
pain pathways, with levels dropping during an attack.
In this WEEKLY, we’ll review the science behind migraines
and explain how Amgen’s new drug works. • Hormonal Link: Migraines are more common in
women than men, so a hormonal link may be tied to
ATTACK OF THE MIGRAINE the causality. There is often a reduction in symptoms
after menopause.
As many as 36 million Americans suffer from
migraines—about 12% of the population. More than • Glutamate Accumulation: In recent years, a few
just a headache, migraines often include symptoms like gene variations that appear to increase the risk
intense pain, nausea, and extreme sensitivity to light or of developing migraines have been identified
noise. They can last anywhere from a few hours to days through genome-wide association studies. Two
on end. Episodes may be as frequent as several times a of these genes result in increased levels of the
month, or as infrequent as a few times a year. neurotransmitter glutamate, suggesting that
accumulation of glutamate in synapses may be
There are three distinct parts of a migraine episode;
a trigger.
however, not all migraine sufferers experience these
phases with the same intensity. Each set of symptoms is
THIS JUST IN
unique to the individual, and can include prodome, aura,
or postdome phases. The hot new kid on the block is calcitonin gene
related peptide (CGRP) inhibitors. CGRP spikes during
• Prodome occurs in the hours or days before a
migraine attacks and is thought to play a role in the
migraine attack. It includes mood disturbances, stiff
brain pathways that process pain. The exact molecular
muscles, and sensitivity to smells or noise.
mechanism of how the CGRP spike is related to migraine
• Aura is the period just before the severe pain onset is not yet fully understood, but CGRP is thought
attacks. Visual distortions are the most common to sensitize nerves in the face, neck, and jaw, as well
symptom, with sensory or motor disturbances as alongside blood vessels surrounding the brain. A
potentially occurring as well. 2002 study provided strong evidence of a key role for
• Postdome happens after the actual headache CGRP in driving migraines. Injecting volunteers who
and includes symptoms such as lingering pain and were migraine-prone with the CGRP peptide induced a
cognitive difficulties. migraine within hours; injecting volunteers who were not
migraine-prone resulted in a mild headache at worst.
THE SCIENCE BEHIND THE EPISODE These studies helped to form the scientific basis for
Amgen’s new monoclonal antibody (mAb) drug that
While the exact cause is largely unknown, there are a
blocks activation of the CGRP receptor. The antibody
few theories:

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binds the receptor but does not activate it, and in so COCKTAIL FODDER: A
doing, prevents CGRP from activating the receptor. HEADACHE OF THE PAST
Amgen may have garnered the first CGRP inhibitor We sometimes blame migraines on the stress of
approval, but several other companies are also working modern living. In some cases, stress is a trigger, but
on drugs that target CGRP. Eli Lilly (Indianapolis, IN) and the headaches are hardly a modern phenomenon.
Teva (Petah Tikva, Israel) both have filed for regulatory Descriptions consistent with migraines are found in the
approval of mAb inhibitors directed at CGRP itself; Alder ancient Egyptian medical text Ebers Papyrus, dating
Pharmaceuticals (South Bothell, WA) has a CGRP- from 1550 B.C., as well as Hippocratic texts dating from
directed mAb in Phase III clinical testing. The idea is for 200 BC.
the mAb to “mop up” CGRP before it reaches the receptor
and triggers a migraine. Allergan (Dublin, Ireland) has a
small molecule inhibitor of the CGRP receptor in Phase III
clinical testing.

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RARE DISEASE FOCUS: PKU Life Science Training from Industry Experts

BIOPHARMA TO THE RESCUE: PKU day one, any infant who tests positive must be fed a
highly restrictive diet. That’s because high levels of
The ubiquitous soda can. Who hasn’t seen one?
phenylalanine can cause developmental delays, irregular
Ever look on the back, at the disturbingly long
motor functioning, and mental retardation. Infant
paragraph of ingredients? The list of ingredients on
diets may contain limited amounts of breast milk, but
the back of a can of diet soda are perhaps even more
rely on phenylalanine-free baby formula. Pediatricians
unsettling. Underneath it, there’s a warning in bold:
recommend pasta, fruits and vegetables, and low protein
“Phenylketonurics: Contains Phenylalanine.”
bread when affected babies start eating solid food.
Phenylalanine doesn’t harm most people. But what’s Lifelong nutritional supplements are key because the
the story with phenylketonurics? They are born with PKU diet lacks protein.
a rare genetic disorder called phenylketonuria. That’s
Scientists still don’t know the exact mechanism
fen-ul-key-toe-NU-ree-uh. More widely known as PKU,
behind PKU. However, its manifestations are clear.
this condition results in an inability to break down the
Images of patients’ brains show damage to both their
amino acid phenylalanine. Untreated, PKU sufferers
white (myelinated axons) and grey matter (neuronal
can face dire health problems including brain damage
cell bodies).
and seizures.
There are two different disease pathways:
To help control the condition, phenylketonurics must
eat a severely restricted diet, limiting intake of high- • Classical PKU is caused by any one of several
protein foods such as meat, dairy, fish, nuts, beans, possible mutations in the gene coding for
legumes, and tofu - all of which contain high levels phenylalanine hydroxylase (PAH). This enzyme
of phenylalanine. Getting back to the diet soda, PKU breaks down phenylalanine. Different mutations
patients can’t consume sweeteners like Equal or affect people differently—some phenylketonurics
NutraSweet, which contain aspartame—a derivative can’t metabolize phenylalanine at all while others
of phenylalanine. experience only diminished ability.
• PKU can also result from a deficiency in the PAH
BREAKTHROUGH cofactor tetrahydrobiopterin (THB). Cofactors
At long last, there’s encouraging news on the subject. are chemical compounds essential to an
On May 24th, the FDA approved BioMarin’s (San Rafael, enzyme’s activity.
CA) latest enzyme replacement therapy, Palynziq.
Developed to treat PKU, the drug promises a more THE NEW PAL FOR PKU PATIENTS
normal life for people unable to savor the simple Palynziq works by replacing PAH, enabling a patient
pleasure of sharing a meal with friends and loved ones. to metabolize phenylalanine. The new drug, taken by
This Biotech Primer WEEKLY examines PKU more closely injection, is a recombinant version of phenylalanine
and how Palynziq can treat it. ammonia lyase (PAL). Related to PAH, this enzyme breaks
down phenylalanine. In clinical trials, people treated
PKU UP CLOSE with Palynziq showed substantially reduced levels of
Phenylketonuria occurs most often in European phenylalanine compared to those on placebo without
Caucasions and their descendants, with a prevalence following the unyielding PKU diet.
of about 1:10,000 births. The condition is autosomal BioMarin already had one drug on the market to treat
recessive—meaning phenylketonurics have two PKU. Kuvan, a small molecule drug, is the synthetic form
defective copies of the gene. Healthy parents often have of THB. It’s taken orally and reduces blood phenylalanine
no idea if they carry the gene in question; consequently, levels in 30% to 50% of patients. Kuvan is indicated for
newborns are routinely tested for PKU. Starting from phenylketonurics whose disease primarily results from

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THB deficiency or certain PAH mutations. While they stem cells, marked for red blood cell formation,
benefit from Kuvan, these patients must still adhere to a are programmed to produce an enzyme that
restrictive diet. breaks down phenylalanine. These phenylalanine-
metabolizing red blood cells are grown in the
COMING UP lab and transfused into the patient where
• BioMarin plans to attack PKU from yet another angle: they remain in circulation for several months.
Gene therapy. Still in preclinical development, this This platform is still in preclinical testing but is
approach would deliver a gene that codes for the generating a lot of excitement within research and
PAH enzyme directly to the liver cells (where it is investment communities.
normally produced) of PKU patients.
COCKTAIL FODDER: TOXIC FUNGUS?
• Synlogic (Cambridge, MA) is taking a novel platform
approach--synthetic biotics. Researchers plan Why does the defective PAH gene persist in human
to engineer gut microbes—healthy bacteria—to populations, when it can be so devastating? It’s thought
produce phenylalanine-digesting enzymes. They that heterozygotes—people with one functional copy
hope to create microorganisms that sense changes and one dysfunctional copy of the gene—have some
in phenylalanine concentrations and respond protection against the effects of one of the most
appropriately. Ultimately, the scientists hope to abundant food contaminants, fungal toxin ochratoxin A.
create a “living drug” that patients can take orally The fungus can occur in grain, pork, coffee, and grapes.
and integrate into their existing gut microbiome. It can also hang out in water damaged houses and
Synologic’s PKU treatment is currently in IND- heating ducts. Turns out, having one copy of the gene
enabling studies. may have helped some of our ancestors survive and
thrive long enough for us to enjoy this edition of Biotech
• Rubius Therapeutics (Cambridge, MA) has
Primer WEEKLY.
announced plans to use its novel red blood cell
therapeutics platform to tackle PKU. Bone marrow

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RED BLOOD CELLS: READY FOR DOUBLE-DUTY? Life Science Training from Industry Experts

RED BLOOD CELLS: READY RBC, transient pores are created, through which the
FOR DOUBLE-DUTY? therapeutic cargo enters.

Biotech Primer WEEKLY talks a lot about white blood • Genetic engineering: Here, researchers modify
cells, with good reason. These powerful immune cells RBCs to contain a gene that codes for a therapeutic
defend us against pathogens and have recently been protein, which the cells then produce.
adapted to fight cancer as CAR-T cells. What about the
body’s other major type of blood cell--red blood cells
THE ROSY FUTURE OF RBC THERAPIES
(RBCs)? Although they receive less media attention, While there aren’t any RBC therapeutics on the market
scientists have long recognized these cells’ critical now, a few are in clinical development:
importance in supplying oxygen to the body. Now drug • Erydel (Urbino, Italy) is developing a treatment
companies are hoping to tap RBCs’ potential to transport platform call EryDex System is which RBCs are
drugs as well. This WEEKLY explains how. loaded up with a drug and infused into patients.
The drug slowly diffuses out of the RBCs over an
RED HOT BENEFITS extended period of time. The loaded-up cells are
RBCs have a few features that make them ideal for referred to as EryDex System End Product (EDS-
delivering drugs: EP). An EDS-EP loaded with dexamethasone sodium
• Stability: These cells live for up to three to phosphate, an anti-inflammatory steroid drug, is
four months. in Phase III clinical testing for the rare inherited
disorder ataxia-telangiectasia. Previous studies
• Biodegradability: After a few months in circulation,
suggest that this drug improves neurological
RBCs are naturally broken down in the spleen and
symptoms in patients.
liver and excreted by the kidneys.
• ERYtech Pharma (Lyon, France) has developed the
• Immune System Tolerance: Assuming a compatible
ERYCAPS RBC-based delivery platform, which, like
donor, red blood cells don’t provoke an immune
EryDel, is based on hypotonic loading of therapeutic
response, significantly reducing the chance of the
agents. Their lead product, Eryaspase, consists of
patient’s body rejecting the product. Red blood cells
RBCs carrying an enzyme called L-asparaginase for
from universal donors– O- and Rh-negative – may
treating cancer. How does it work? L-aspariginase
serve as a starting point for “off-the-shelf” therapies.
breaks down the amino acid asparagine. Healthy
These can be prepared in advance for multiple
cells can synthesize this amino acid, whereas many
patients, rather than custom-made for each patient.
types of cancer cells can’t. By encapsulating the
Given these characteristics, how do researchers turn red enzyme in long-lived RBCs, it stays in circulation
blood cells into drug delivery systems? long enough to degrade available asparagine.
• Attachment: Fixing therapeutic proteins, small This “starves” the cancer cells that can’t make
molecule drugs, or nanoparticles to a cell’s surface it themselves. Eryaspase is now in Phase III
creates “carrier RBCs.” In this approach, drugs clinical studies for acute lymphoblastic leukemia
are chemically linked to the RBC or attached to and in Phase II for acute myeloid leukemia and
antibodies that then bind to the RBC surface. pancreatic cancer.

• “Loading” red blood cells: Researchers place the

THE NEXT BIG THING
cells in a solution with a lower concentration of
IN IMMUNOTHERAPY?
salts and other materials than that of the red blood
cell. The difference in concentration causes water One of the most interesting areas of preclinical
to move into the cell. As the water moves into the development for RBC therapeutics is immunotherapy:

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 • Cancer: Rubius Therapeutics (Cambridge, MA) is EASILY CONFUSED: ALLOGENIC
genetically engineering RBCs to display proteins VS. AUTOLOGOUS
on their surface that can activate killer T-cells to
Cell therapies such as red blood cell therapies, CAR-T
destroy tumors.
therapies and stem cell therapies are classified as either
• Autoimmune disease: By activating a different allogenic or autologous. Autologous cells come from
type of T-cell, regulatory T-cells, researchers are the patient’s own body. An example is current CAR-T
hoping to defeat autoimmune diseases, including therapies, in which white blood cells are removed from
Type 1 diabetes. The immune systems of patients the patient, modified in the lab to be tumor-seeking,
with this illness attack the insulin-producing cells and infused back into the patient. The advantage of
of the pancreas. Scientists at Anokion (Lausanne, using autologous therapies is there is essentially no risk
Switzerland) are developing a platform that involves of immune system rejection, since the cells come from
attaching portions of “autoantigens” – proteins in the patient’ own body. The downside? They are more
patients’ bodies that induce autoimmunity – to the expensive and time-consuming to produce than allogenic
surface of RBC. This “trains” the patient’s immune therapies, which are derived from cells that come from a
system not to respond to his or her insulin-producing compatible donor and are not patient-specific. Many of
cells. Rubius is also working on genetically- the RBC therapies being tested are allogenic. Scientists
engineered RBCs to induce immune tolerance. are working on developing allogenic CAR-T therapies.

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GLUTEN: PROTEIN OF DOOM Life Science Training from Industry Experts

CELIAC DISEASE TERM OF THE WEEK: PEPTIDE

Imagine the tantalizing scent of freshly baked bread. A peptide is a chain of amino acids consisting of fifty or
Now, think about even a single scrumptious slice fewer amino acids.
wreaking havoc on you: stomach pain, bloating and other Peptides are part of us. Or at least inside us—as
digestive consequences. Welcome to Planet Celiac— mentioned above, they come from protein digestion.
home to the millions of Americans diagnosed with celiac Our bodies also produce and put to work several other
disease. Today’s WEEKLY looks at this autoimmune peptides too. For example, the pancreas pumps out the
disorder and how biotech companies are working to peptide hormone glucagon in response to low blood
make life a lot more comfortable for celiac patients. sugar. Glucagon signals the liver to release stored
glucose into the bloodstream. Other powerful peptides
GLUTEN: PROTEIN OF DOOM include amylin, which slows our stomach’s emptying
Celiac disease is triggered by gluten proteins found in and prolactin, which stimulates milk production in
wheat. Typically, our bodies break down the proteins nursing mothers.
into their chemical building blocks (amino acids). Our
intestines then harmlessly absorb them to make the new EASILY CONFUSED: CELIAC DISEASE
proteins essential to cellular function. VS. GLUTEN SENSITIVITY
Glutens, however, contain unusually high levels of Celiac disease and gluten sensitivity each involve
proline. This amino acid endows glutens with the power the body’s response to peptides derived from gluten
to resist digestive enzymes. The resulting incomplete proteins. However, the details of that response
digestion causes us to produce short chains of amino differs significantly.
acid chains or peptides. In people with celiac disease, Celiac disease patients mount “an antigen-specific
peptides provoke their bodies’ immune systems to immune response.” They generate antibodies that
attack their own small intestine. Ow. recognize gluten peptides and unleash T-cells (a type
Symptoms of celiac disease range from diarrhea, weight of white blood) to fend them off. The trouble is, the
loss, and malnutrition to the less obvious such as overzealous T-cells also hone in on the patient’s own
isolated nutrient deficiencies. Some people with celiac small intestine, often causing significant damage
disease suffer no symptoms at all. and discomfort.
In contrast, gluten sensitive patients launch an “innate
BREAD AND OTHER DREADS immune response” towards gluten. While they don’t
So, gluten is a protein. And very bad for celiac disease produce gluten-specific antibodies or activate gluten-
patients. But what is it? Glutens are the proteins in specific T-cells, their immune system recognizes gluten
wheat, barley and rye. Those grains are in bread, peptides as foreign and potentially harmful, leading to
of course—but in lots of other food and drink too, inflammation that can cause celiac-like symptoms. The
including: cereal, beer, malted milkshakes, and significant takeaway difference is that people with gluten
salad dressing! sensitivity suffer no direct damage to their intestine. The
So, what’s a citizen of Planet Celiac to do? Currently, two disorders can be differentiated by testing for celiac-
the only recourse is totally avoiding food and drink specific antibodies.
containing gluten. It sounds simple perhaps, but gluten
is a tricky foe—salad dressing? Beer? Really? Really. COMING DOWN THE PIKE
Biotech may have a few fixes in store for the more
than twenty million Americans who deal with celiac
disease or gluten-sensitivity. Scientists are approaching

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the problem from two different angles. One, they’re PvP Biologics (San Diego, CA) is developing KumaMax,
working on modifying wheat and other grains so they an enzyme that breaks down gluten in the stomach
don’t kick the body’s immunity into action. And two, before it damages the small intestine. Now in preclinical
they’re exploring ways to treat patients so their immune testing, KumaMax is derived from an enzyme
systems responds more mildly. called kumamolisin. Kumamolisin itself is found in
Eat That Wheat? Alicyclobacillus sendaiensis, a type of bacteria that that
grows in acidic conditions – similar to those found in the
Wheat glutens contain two proteins: gliadins and
stomach, meaning that the enzyme can be delivered
glutenins. Gliadins give rise to the problematic peptides
orally and should be able to function efficiently in the
and rev up the immune system. Researchers from the
stomachs of celiac patients.
Spanish Institute for Sustainable Agriculture used
CRISPR/Cas9 genome editing to develop a strain of COCKTAIL FODDER: X-TREMOPHILES!
wheat without gliadin proteins. The new-fangled wheat
Alicyclobacillus sendaiensis is an extremophile. These
scored high marks from a panel of trained assessors
bacteria that grow in—you guessed it--extreme
when compared to traditional wheat. Small-scale trials
environments. Humans and most plants, animals
assessing safety in celiac patients are now being carried
and fungi have adapted to a very limited range of
out in Spain and Mexico.
conditions such as body temperature, pH (acid level),
Taming the Celiac Response and atmospheric pressure. Extremophiles have instead
ImmusanT (Cambridge, MA) is developing a evolved to survive crazy environments. One of the most
desensitization therapy in which patients are famous extremophiles you’ve probably never heard of
systematically exposed to peptides that elicit the is the Taq polymerase. All sorts of biotech use Taq in
celiac disease immune response. Researchers hope the polymerase chain reaction (PCR). This reaction uses
to “reprogram” T-cells to no longer freak out over a DNA-copying enzyme to amplify DNA and requires
the problematic peptides. This approach echoes temperatures near boiling. Most DNA-copying enzymes
desensitization therapies for allergies such as pollen, can’t handle the heat. In contrast, Taq is literally hot
pet dander, and dust mites. Once considered too risky stuff. Taq’s affinity for steamy temperatures isn’t
for food allergies, new studies support the benefits of surprising. After all, its bacterial “parent” came from the
gradual exposure to food allergens. ImmusanT’s product, hot springs at Yellowstone National Park.
Nexvax2, has completed Phase 1b studies. Preliminary
data suggests that it may increase celiac sufferers’
gluten tolerance.

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BREAKING BAD WITH SCLC & NSCLC Life Science Training from Industry Experts

BREAKING DOWN LUNG CANCER decay of radioactive elements occurring naturally in soil
and rocks. Radon is not considered dangerous because
The hit TV series Breaking Bad features anti-hero
it is usually present at very low levels. However, it can
Walter White, who starts out as a sympathetic
sometimes build up to dangerous levels in well-insulated,
character: a mild-mannered high school chemistry
tightly-sealed homes built on soil rich in uranium,
teacher with a nagging cough that turns out to be
thorium, or radium.
lung cancer. Money problems precipitated by costly
treatments, poor insurance, and a modest salary push Asbestos
him to start cooking up meth to ensure the financial Asbestos used to be a common insulating material
security of his family. Spoiler alert: The treatments used in buildings and ships. The microscopic fibers in
succeed beyond his expectations, restoring his health asbestos can be inhaled and become lodged in lung
long enough for him to become an unexpected cells, triggering the activation of inflammatory pathways
meth kingpin. that result in the release of mutagens and factors that
Breaking Bad is a fictionally extreme example of the promote tumor growth. Since its hazards became
chaos that can arise from a lung cancer diagnosis. In well-documented in the mid-1970s, it is no longer used
fact, lung cancer is the leading cause of cancer-related as insulation.
deaths in the United States. Let’s take a closer look at the Genetics
molecular causes, the different types, and some of the In addition to carcinogen exposure, there are likely
treatments in the clinic and on the market. genetic elements that make certain individuals more
or less susceptible to lung cancer. Even though 90% of
THE DANGER lung cancer cases are caused by smoking, only about
While Walter White did not smoke cigarettes, 90% of 10% of smokers get lung cancer. In African-American
those affected by lung cancer are smokers. Other causes populations, even when differences in smoking rates and
of lung cancer include environmental or workplace access to healthcare are controlled for, the rates of lung
exposure to carcinogens (known cancer-causing agents) cancer are higher. Both of these scenarios suggest that
such as radon, asbestos, or air pollution. there may be genetic factors that make certain people
Smoking more (or less) susceptible.

Smoking causes cancer because the inhaled smoke

SMALL CELL LUNG CANCER
contains a range of chemicals, many of which are known
to be carcinogens, including benzene, formaldehyde, About 10% of lung cancer is small cell, meaning it occurs
methanol, and acetylene. Some carcinogens are in the very small cells found in the bronchii—the tubes
genotoxic, meaning that they cause cancer by directly that branch off of the trachea, enter the lungs, and
interacting with and damaging DNA. If that DNA damage divide into even smaller branches within the air sac.
occurs in a gene involved in regulating cell division, There are currently no targeted therapies available for
cancer may result. Non-genotoxic carcinogens have no small cell lung cancer (SCLC), with chemotherapy and/
direct interaction with DNA; rather, they disrupt cellular or radiation as the main line of treatment. Therapies in
structures and change the rate of either cell division or development for SCLC include:
processes that increase the rate of genetic error. • Immune checkpoint inhibitor therapies Opdivo
Radon Gas (Bristol Myers Squibb; New York City, NY) and
Radon gas exposure can result in cancer because it is Keytruda (Merck; Kenilworth, NJ). Checkpoint
radioactive, and the high-energy radioactive particles proteins are proteins on the surface of white blood
given off as the gas decays can cause direct damage to cells such as T-cells that send a “stop attacking”
cellular DNA. Radon gas is released from the normal signal when activated by proteins on the surface

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 of healthy cells in the body. Many types of cancer 35% of NSCLC cases are caused by the over-expression
cells take advantage of this system by activating of the growth factor receptor EGFR. These types of
the immune checkpoint proteins and minimizing NSCLC—more common in non-smokers—can be treated
the immune response against a tumor. Checkpoint by drugs that target and inhibit this receptor. These
inhibitor therapies prevent the cancer from include Iressa (Astra Zeneca; London, UK), Vectibix
activating checkpoint proteins, enabling the immune (Amgen; Thousand Oaks, CA), Tarceva (Roche; Basel,
system to more fully go after the tumor. Both Switzerland), and Afatinib (Boehringer Ingelheim;
drugs are being tested as monotherapies and in Ingelheim, Germany). Pfizer (New York, NY) has recently
combination with other drugs. Opdivo was granted announced positive Phase III results of dacomitinib,
an FDA Priority Review for the treatment of SCLC, a new small molecule inhibitor of EGFR, while Rain
with an approval decision expected by August 16th Therapeutics (Fremont, CA) has just completed Phase
2018. Keytruda is in Phase III clinical testing for SCLC. I trials of Tarlox, an EGFR-inhibitor for patients who do
• Rova-T (AbbVie; North Chicago, IL), now in Phase III not respond to those already on the market.
clinical testing, is an antibody-drug conjugate that About 5% of NSLC cases are caused by mutations in a
delivers a toxic chemotherapeutic agent directly gene known as anaplastic lymphoma kinase (ALK). ALK
to SCLC cells. Rova-T is being studied as both a proteins activate cell division, and mutated versions can
monotherapy and in combination with Opdivo. drive cell division inappropriately. The drugs Xalkori
• Aeglea Biotherapeutics (Austin, TX) is developing (Pfizer; New York City, NY) and Zykadia (Novartis;
Pegzilarginase, a therapeutic enzyme meant Basel, Switzerland) inhibit ALK. Pfizer is developing
to starve small cell lung cancer cells by breaking lorlatinib, another ALK inhibitor, for patients who
down the amino acid asparagine. Healthy cells can have become resistant to Xalkori; Ignyta (San Diego,
synthesize this amino acid, whereas SCLC cells can’t. CA) also has a new ALK-inhibitor, entrectinib, in
Pegzilarginase is now in Phase I clinical studies clinical development.
and is being tested as both a monotherapy and in Opdivo and Keytruda have also been approved for use in
combination with Keytruda. NSCLC patients.

NON-SMALL CELL LUNG CANCER COCKTAIL FODDER:

Cancer that occurs within any cell outside of small cells is WALTER’S DIAGNOSIS
referred to as non-small cell lung cancer (NSCLC), making Diagnosed with NSCLC, Walter White specifically had an
up the majority (~90%) of lung cancer cases. inoperable stage 3A adenocarcinoma. This means the
A number of drugs targeting new blood vessel growth— cancer was initiated in the mucus-producing cells of the
angiogenesis inhibitors—have been approved for the lungs and had spread to the lymph nodes (or other sites
treatment of NSCLC. These include Avastin(Genentech; near the lungs) but had not spread to distant sites within
South San Francisco, CA) and Cyramza (Eli Lilly; the body. Some types of adenocarcinomas are caused by
Indianapolis, Indiana). These drugs cut off the ALK mutations, so it is possible that Walter’s miraculous
supply of new blood vessels to the tumor, essentially recovery was caused by one of the ALK inhibitors
starving them. discussed above.

There are also drugs that target specific NSCLC–

associated mutations. For example, between 10% and

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PHAGE THERAPY: NEW HOPE FOR
ANTIBIOTIC-RESISTANT BACTERIA Life Science Training from Industry Experts

OLD WAY OF FIGHTING coexisted with bacteriophage for a long time, suggesting
BACTERIA RENEWED that there should be few safety issues with their use
as therapeutics.
One of the greatest public health challenges of the 21st
century is antibiotic resistance, which occurs when a Scientists have known about the bacteria-devouring
few bacteria in a given population develop a genetic ability of phage for about 100 years, but with the advent
mutation that enables them to survive — even in the of antibiotics in the late 1920s, medicine’s focus shifted
presence of antibiotics.How do bacteria become drug to these new wonder drugs because they were easier to
resistant? Suppose a particular antibiotic inhibits manufacture and test in controlled settings. Now that
an enzyme required for bacterial replication. If one antibiotic resistance is emerging, so, too, is a renewed
bacterium mutates so the enzyme has a slightly different interest in bacteriophage, which are now starting to
shape, the antibiotic is no longer effective. The mutated be manufactured and tested in a standardized way for
bacterium lives on and continues to replicate, even as the first time. Let’s take a look at some of the biotech
all the others die off. Over time, this resistant strain companies delving into the world of bacteriophage-
becomes dominant, spreading from person to person, based therapeutics.
remaining unchecked and thriving. It is not uncommon
for a strain of bacteria to become resistant to several
THE COCKTAIL APPROACH
different antibiotics, giving rise to the term multi-drug The first multicenter clinical trial examining the use
resistant bacteria.In this issue of the WEEKLY, we’ll take a of bacteriophage as antibacterial treatments was
look at a novel approach to fighting bacterial infections initiated in 2015 by French biotech Pherecydes (Paris,
— bacteriophage. France). Preparative work for the trial began in 2013 as
researchers established protocols for producing phage
TERM OF THE WEEK: BACTERIOPHAGE that met good manufacturing practice guidelines. The
A bacteriophage — also referred to as a phage — is a researchers are studying two different “cocktails” of
virus that infects bacteria. By attaching to a bacterium’s bacteriophage — mixtures of different bacteriophage
surface, a phage punches holes in the membrane and that have shown activity against different substrains
injects its own genetic material inside. The phage then of a particular bacteria in the lab. The first contains 13
replicates inside of the bacterium, creating so many new different phages targeting P. aeruginosa; the second, 12
viruses that the bacterium breaks open, releasing newly phages that target E. coli. Both are being evaluated for
produced viruses, which can then go on to infect other the treatment of burn wound-associated infections. The
bacteria, continuing the cycle. company is also preparing to begin clinical trials of phage
therapy for the treatment of diabetic foot ulcers infected
The word “bacteriophage” is derived from the Greek
by S. aureus.
word phagein — “to devour.” So we can think of
bacteriophage as, literally, devouring bacteria — a Other companies testing phage cocktails in human
potentially very useful trait! Typically each phage is patients include:
specific for a type of bacteria, meaning that if adapted • AmpliPhi Biosciences (Richmond, VA): Phage
for therapeutic use, researchers can select viruses cocktail AB-SA01 has completed Phase I clinical
that will only attack harmful bacteria, and leave the testing for antibiotic resistant S. aureus in two
many strains of “friendly” bacteria that make up our different clinical settings: Chronic rhinosinusitis
gut microbiome alone. And since each type of phage as well as acute and chronic wound and skin
has coevolved for millennia with its chosen strain of infections. Additional target indications include
bacteria, each adapting and changing in response to the bacteremia, endocarditis, prosthetic joint infections,
other, resistance is much less likely to evolve as has been osteomyelitis, and diabetic foot ulcers.
the case for antibiotics. Likewise, humans have safely

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 • TechnoPhage (Lisbon, Portugal): Phage cocktail TP- the bacteriophage’s antibacterial response even
102 is in Phase I studies targeting bacteria associated stronger. Companies with engineered bacteriophage in
with chronic ulcers. preclinical development include Synthetic Genomics
• Intralytix (Baltimore, MD): Completed Phase I (San Diego, CA), C3J Therapeutics (Los Angeles, CA), and
studies using a bacteriophage for the treatment of EnBiotix (Cambridge, MA).
infected wounds. Recently announced that they have
received FDA clearance to begin Phase I/IIa testing of
THE COMPONENT APPROACH
phage therapy for inflammatory bowel disease. A third approach to tapping into the therapeutic power
of bacteriophage lies in isolating the component or
• EpiBiome (South San Francisco, CA): Phage cocktails
components that make them toxic to bacteria. For
targeting E. coli and S. dysenteriae diarrheal infections
example, in order to inject their genome into bacteria,
in preclinical development.
phage must essentially punch a hole in the membrane,
THE ENGINEERED APPROACH which is itself very damaging to the bacteria. The viral
protein that creates these tears in the membrane
Creating a phage cocktail can be a challenging
are called “lysins” — enzymes that essentially chew
biomanufacturing problem. An alternative approach to
holes in the bacterial cell wall. ContraFect (Yonkers,
combining beneficial characteristics of different viruses
NY) is conducting Phase II clinical studies of its drug
is to genetically engineer a synthetic virus that combines
CF-301 — a lysin — for the treatment of S. aureus
the properties of multiple phages into a single genome.
bloodstream infections.
For example, scientists could insert genes into a phage
Although still in the early days of clinical testing,
genome to increase the range of bacteria subtypes
bacteriophage offer the possibility of being a safe and
an individual phage can attack, yet still maintain the
effective solution to the current antibiotic resistance
specificity that prevents the phage from raiding friendly
public health crisis.
bacteria. Researchers could also add in genes to make

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THE IMMUNE SYSTEM: FRIEND
& SOMETIMES FOE Life Science Training from Industry Experts

THE IMMUNE SYSTEM: FRIEND Macrophages are one type of non-specific defender.
& SOMETIMES FOE When they encounter a virus or bacteria, they engulf and
destroy it – think Pac-Man. Mmm---germs! Macrophages
The immune system. Yes, everybody has one. It’s pretty
can differentiate between benign and harmful
important to our health. But what the heck is it? In short,
microorganisms thanks to a special receptor. Called a
the immune system is a complicated network of organs,
Pathogen-Associated Molecular Pattern, or PAMP,
cells, and signaling molecules. Without this guardian, we
they’re found only on microorganisms that make us sick.
fall prey to dangerous pathogens and toxic substances in
the environment. The immune system can even protect Other non-specific defenders include neutrophils and
us from harmful changes that occur within our bodies. natural killer cells. Neutrophils also recognize PAMPs
With the exception of the nervous system, no other and engulf invaders. Natural killers inject the protein
system in the body is as complex. Every day, biopharma granzyme B into invaders, which eradicates them.
researchers apply what they know about human Once activated, non-specific defenders release
immunity to treat disease. inflammatory cytokines. These signaling molecules
switch on other immune cells. The resulting
NOW HOW DOES THIS THING WORK? inflammatory cascade helps ensure a rapid,
The immune system protects us from harmful invaders, comprehensive response.
including viruses, bacteria, parasites, and fungi. It may
also recognize threats from within, such as cancer. The SPECIFIC IMMUNITY: T-CELLS & B-CELLS
immune system has many components, working in When non-specific defenses flop, it’s time to call in the
concert for our health. The first defenses are physical cavalry — T-cells and B-cells. These highly-specialized
barriers—such as our skin, mucus membranes, and cells make up specific (or adaptive) immunity. These
the tiny hairs that line our upper respiratory tract. The cells are so specialized, each individual T- or B- cell only
barriers work to stop pathogens from getting in at all. recognizes one unique target - usually a specific protein
When they fail, our cellular defense mechanisms kick in. on the surface of an invading pathogen. This is possible
As with so many other aspects of human nature, the due to their uniquely-shaped cell-surface receptor
immune system can be paradoxical. Our “bodyguard” proteins. Depending on the exact shape of the receptor,
fights illness and disease. But sometimes it goes haywire. a different target protein is recognized. When a given
When that happens, the immune system attacks us. This receptor recognizes and binds its target, the associated
overactivity forms the basis for autoimmune disorders, T- or B- cell is activated. What happens next depends on
such as Crohn’s disease and rheumatoid arthritis. More the type of cell we are talking about.
on that later.

NON-SPECIFIC IMMUNITY:
SEARCH & DESTROY
Our amazing bodies possess two kinds of immunity:
Non-specific or innate and specific or adaptive. At the
core of both of these responses are specialized white
blood cells that recognize and attack foreign invaders.
Non-specific immunity is our first line of defense against
invading pathogens. Most white blood cells are non-
specific defenders, meaning they attack in the same
Activated T-cells divide rapidly and produce three
fashion regardless of a threat’s specific characteristics.
distinct descendants: killer T-cells, helper T-cells, and

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memory T-cells. The “kids” all recognize the same target white blood cells to attack a tumor. Chimeric antigen
as does their parent. receptor T-cell (CAR-T) therapies are T-cells whose
• Killer T-cells roam the body, searching for their receptors have been engineered to recognize and
preprogrammed epitopes. When they find one, they destroy cancer.
jab it with granzyme B, triggering cell death.
FOILING THE CHECKPOINT IMPOSTERS
• Helper T-cells release inflammatory cytokines
Our body has a natural screening process — checkpoints
that galvanize antibody-producing B-cells, killer
to prevent autoimmune disorders. They stop T-cells and
T-cells, and macrophages en masse. The human
B-cells from assaulting our tissues. Checkpoint proteins
immunodeficiency virus (HIV) infects only helper
send a cease and desist signal to T-cells when they
T-cells. In so doing, the virus completely cripples the
encounter healthy cells. Many types of cancer actually
immune response.
make more than normal levels of proteins that activate
• Memory T-cells don’t defend against initial infection. these checkpoints, “tricking” the adaptive immune
But —- If they encounter the same epitope a second system into ignoring malignant cells. Checkpoint
time, they quickly morph into killer T-cells and helper inhibitor therapies prevent cancer cells from triggering
T-cells, which then take out the pathogen. Memory these checkpoints. This enables the immune system to
cells form the basis of immunity, quickly jumping in work against the tumor.
to defend against a recognized virus.
Activated B-cells also reproduce rapidly and produce AUTOIMMUNE DISEASE: OUR
two types of descendants: plasma cells and memory FRENEMY, THE IMMUNE SYSTEM
B-cells. Our immune system is our friend, mostly. Still, an
• Plasma cells secrete antibodies – proteins that overactive immune system can cause serious problems,
recognize and bind to bacterium or virally-infected such as autoimmune disease. Chronic inflammatory
cell that sports a matching epitope. The binding disorders, such as Crohn’s disease, rheumatoid
triggers other immune cells, such as killer T-cells or arthritis, and psoriasis wreak havoc by provoking
macrophages, to destroy the pathogen. white blood cells to target healthy cells and release
inflammatory cytokines.
• Like memory T-cells, memory B-cells lie dormant,
waiting to defend the body against the same Biologic drugs that treat autoimmune disorders—like
foreign invader. Humira (AbbVie, North Chicago, IL), Enbrel (Amgen,
Thousand Oaks, CA), and Rituxan (Genentech)—shut
IMMUNOLOGY TO THE RESCUE down key parts of the immune response. Humira
The biotech industry has elegantly hacked the immune and Enbrel inhibit a specific inflammatory cytokine,
system to make important immunotherapies that use TNF-alpha. These drugs are approved for a range
strategies derived from specific immunity. Scientists of inflammatory diseases. Rituxan, approved for
have developed monoclonal antibody therapies by rheumatoid arthritis, reduces the number of B-cells that
selecting antibodies that recognize and bind to a target the joints.
disease-specific protein. A great example of this is Be it our body’s friend or frenemy, the immune system
Herceptin (Genentech; South San Francisco, CA). This continues to challenge and reward scientists as they
lab-born antibody binds to the HER2 growth-factor delve into its potential to improve human health. Next
receptor, which occurs in high levels on one quarter week, we’ll look more closely at antibody therapeutics,
of breast cancer patients’ tumors. Herceptin mimics exploring the latest innovations.
natural antibodies, compelling killer T-cells and other

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AMAZING ANTIBODIES PART ONE: BISPECIFICS
& ANTIBODY-DRUG CONJUGATES Life Science Training from Industry Experts

ATTACK OF THE MONO- & POLYCLONALS scientists may identify an antibody that binds to a
cancer cell antigen. When the antibody is injected into
Monoclonal antibody (mAb) therapeutics burst onto the
a patient, this binding prods the patient’s immune
healthcare scene twenty years ago. They remain one
system to attack the cancer. This is the mechanism of
of the most versatile and effective therapies available
action used by Herceptin (Roche; Basel, Switzerland)
for a whole range of diseases including different types
to attack HER2-positive breast cancer, and by Rituxan
of cancers, autoimmune diseases, infectious diseases,
(Roche) to attack non-Hodgkin’s lymphoma and chronic
and most recently, even high cholesterol. Tried and true
lymphocytic leukemia.
mAbs, such as Herceptin and Rituxan, remain in high
demand. More on those below. EASILY CONFUSED: MONOCLONAL
Biotech companies have built on the success of this first VS. POLYCLONAL ANTIBODIES
wave of mAbs to develop new, innovative products that Antibodies come in two types. Monoclonal antibodies
deliver an even bigger punch. This WEEKLY reviews the all derive from the same B-cell, or its clones
basics of monoclonal antibodies and highlights some (descendants). This enables them to recognize the same
recent innovations. epitope. Therapeutic antibodies are always monoclonal,
which ensures consistent treatment.
JUST THE FACTS
Polyclonal antibodies are produced by a collection of
• Antibodies are proteins produced by B-cells, a type
different B-cells. As a result, they recognize multiple
of white blood cell. An antibody’s shape corresponds
epitopes on the same antigen, which makes them ideal
to a unique target, typically a protein on the surface
for some kinds of diagnostics and research, where the
of a virus or other pathogen. Scientists call these
only requirement is detecting a specific antigen.
foreign proteins antigens.
• When an antibody recognizes its particular antigen, NEW & IMPROVED Y
it binds to it. The action alerts the attack cells of the Fun fact: antibodies are Y-shaped. In antibodies
immune system such as macrophages and killer produced naturally by our bodies and in most
T-cells, which then eliminate the pathogen. therapeutic antibodies, the two “arms” of the Y are
• The bit of the antigen the antibody locks onto is identical and recognize only one target.
an epitope. In contrast, bispecific antibodies have been genetically
engineered by splicing genes for two different
monoclonal antibodies to make a new Y. This way, the
bispecific antibody is able to recognize two different
targets and bring them in contact with one another.
Imagine that one arm of the Y recognizes a cancer cell.
Meanwhile, the other arm recognizes and binds to a killer
T-cell. Remember, killer T’s are white cells that inject
toxins directly into cells. By bringing the malignant cell
it “caught” into contact with a killer T-cell, the first arm
of the Y essentially forces the killer-T-cell into action —
killing the cancer cell.
Antibodies’ amazing ability to recognize and bind
The cool thing is that it’s not even necessary to imagine
to unique epitopes makes them highly effective
this scenario — it’s the mechanism that Blincyto (Amgen;
therapeutics. Researchers have capitalized on this
Thousand Oaks, CA) uses.
“bindability” in a number of ways. For example,

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 • A highly toxic small molecule drug to kill the cancer
cell once internalized.
• A chemical linker that connects the small molecule
drug to the antibody. The linker is stable in a
person’s bloodstream, releasing the drug once inside
the tumor.
How does this work?
1. The antibody binds to its target antigen on the
cancer cell surface.
2. The antibody-drug conjugate is then taken up -
internalized - by the cell.
So far only two bispecific antibodies have been approved
by the FDA: Blincyto and Roche’s (Basel, Switzerland) 3. Once inside the malignant cell, the linker degrades
Hemlibra, approved to treat hemophilia A. Hemlibra and the active drug is released.
works by bringing together blood clotting factors IX and The ability to target only cancer cells allows doctors
X, enabling the activation of factor X. In healthy people, to administer much more toxic medicines than
factor X is activated by clotting factor VIII, but hemophilia with traditional chemotherapy. This is because the
A patients lack this factor. ADC’s precision means it avoids healthy tissue that
chemotherapy often damages or destroys.
THE ABCS OF ADCS
There are currently four ADCs on the market:
Another type of game-changing innovation in mAb
• Seattle Genetics’ (Seattle, WA) Adcetris for
technology is antibody-drug conjugates (ADCs): highly-
Hodgkin’s lymphoma
potent, targeted therapeutics that combine the targeting
power of monoclonal antibodies with the cancer-killing • Roche’s Kadcyla for HER2-positive breast cancer
ability of toxic drugs. This potent combo can destroy • Pfizer’s (New York, NY) Besponsa for acute
cancer cells with less impact on healthy cells. lymphoblastic leukemia
ADCs have three key components: • Mylotarg, also by Pfizer, for acute myeloid leukemia.
• A monoclonal antibody that is highly specific for a The next WEEKLY continues our foray into
tumor-associated antigen with little to no expression next-generation antibodies with a look at
on healthy cells. photoimmunotherapy and nanobodies.

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AMAZING ANTIBODIES PART TWO:
ENLIGHTENED & NANO Life Science Training from Industry Experts

WHAT CAN’T THESE LITTLE DUDES DO? the blood, like inflammatory cytokines. Developing cell-
penetrating mAbs would open up a new world of healing.
When last we met, we discussed the fundamentals of
monoclonal (mAb) therapies and looked at two recent Scientists at Ablynx (Ghent, Belgium) are doing just
advances: antibody-drug conjugates and bispecific that. Moreover, their fancy schmancy new mAb derives
antibodies. This week continues our adventure in from a perhaps unlikely source — camels and llamas.
antibody innovation by introducing antibody-based No offense to us, but these camelidae antibodies
photoimmunotherapy and nanobodies. are structurally and functionally very similar to our
antibodies. Happily, they possess a few important
LIGHTENING CANCER differences that may add up to something way bigger
PATIENT’S PROSPECTS than any antibody.

Photoimmunotherapy was conceived of at the A camelidae antibody works, like any other, because
National Institutes of Health and is being developed by its specific shape enables it to recognize and bind to a
Aspyrian Therapeutics (San Diego, CA). This treatment specific pathogen. But — these antibodies are only a
“enlightens” antibodies to amplify their disease fighting tenth the size of those found in mammals. Hence the
ability. Here’s how it works: name “nanobodies” (for you word nerds — from the
Greek word for dwarf: nanos.) Because their small size
1. A light-sensitive, nontoxic drug is attached to the end
allows them to actually enter cells, these miniscule
of an antibody. The drug becomes toxic, that is, “live,”
disease fighters can recognize targets hidden away
when exposed to infrared light.
inside of those cells. Their stature may also enable them
2. A cancer patient is injected with the antibody-light to cross the tricky blood-brain barrier and/or penetrate
sensitive drug combo. The antibody finds its target the interior cells of tumors —– two activities that
antigen on the surface of a cancer cell. conventional antibody therapies lack.
3. The next step is activation. To accomplish this, Nanobodies are less structurally complex than their
infrared light is shone at the tumor, switching sizier relations. Researchers have capitalized on
on the drug, which then damages the cancer cell their relative simplicity by manufacturing them in
membrane. The damage enables water to flood the bacterial cells – something that is not possible for more
cell, which consequently bursts and dies. complex proteins, and which significantly reduces
4. This rupture releases tumor antigens that spark the production costs when compared to the mammalian cell
immune system into recognizing and responding to manufacturing required for traditional antibodies.
more malignant cells. Preliminary studies in mice also suggest it’s possible for
Important caveat: photoimmunotherapy only works nanobodies to survive in our stomach and intestines.
if the infrared laser can reach the tumor and trigger This initial success raises the prospect of oral nanobody
the photosensitive drug. Aspyrian Therapeutic’s lead therapy for conditions such as Crohn’s disease.
candidate ASP-1929 is preparing to enter Phase III clinical
studies to treat head and neck cancers. IN THE CLINIC
Look for the first nanobodies on the market later this
SMALL IS POWERFUL, BELIEVE IT! year. On June 29, the European Medicines Agency’s
Despite their success, mAbs do have a chink in their Committee for Medicinal Products for Human Use
armor. They’re pretty big, cellularly speaking. Their (CHMP) recommended approval for Ablynx’s first
microscopically huge size prevents them from actually nanobody therapeutic, caplacizumab. Ablynx developed
getting into cells. This limits their therapeutic power. For the drug to treat a rare blood-coagulation disorder.
now, mAbs can only target antigens on a cell’s surface, The disease, acquired thrombotic thrombocytopenic
for instance receptor proteins, or those circulating in purpura (aTTP), causes widespread microscopic clots in

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blood vessels. The disease is triggered by too much von on the surface of a common and very contagious virus
Willebrand factor (vWF), a protein that initiates blood — respiratory syncytial virus (RSV). The F protein allows
clotting. caplacizumab inhibits vWF, keeping a patient’s RSV to lock onto lung cells. Ablynx expects ALX-0171 to
blood clot-free. disrupt the interaction between F protein and the lungs,
The company already has two more nanobody products thereby preventing infection.
in Phase II clinical testing: Vobarilizumab and ALX- Since their introduction in the 1990s, monoclonal
0171. Vobarilizumab (say that three times fast) reduces antibodies have shown themselves safe and effective
the activity of the protein interleukin-6 (IL-6), which in many areas. Innovations in this powerful technology,
stimulates immunity. Inhibiting a person’s immune such as photo immunotherapy and nanobody
system may prove useful against autoimmune disorders. development, ensure these tiny components of the
The new drug is being tested in partnership with AbbVie immune system remain one of the biggest tools
(North Chicago IL) for rheumatoid arthritis and lupus. in biopharma.
Next up is ALX-0171, which binds the fusion (F) protein

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PILLS, PEPTIDES, & PROTEINS Life Science Training from Industry Experts

PROMISING PEPTIDE THERAPIES molecule drugs which include sensitivity to digestive

enzymes, delivery by injection, and high specificity
The front runners in the game of drug delivery include
for their target.
small molecule and large molecule drugs, but there is
another class that lands right in between: peptides.
Several companies, including Rhythm Pharmaceuticals
(Boston, MA), Kalos Therapeutics (San Diego, CA),
Aileron Therapeutics (Cambridge, MA), and Bicycle
Therapeutics (Cambridge, MA) have emerged as
prominent players in the peptide arena.
Let’s review the differences between the drug classes
and explain where peptides fit into the picture. Then
we’ll take a spectator’s interest in the companies and
products leading the charge in peptide therapeutics.
Examples of peptide drugs on the market today include
EASILY CONFUSED: SMALL MOLECULE glucagon-like peptide-1 (GLP-1) receptor activators, such
VS. LARGE MOLECULE VS. PEPTIDE as Byetta (AstraZeneca; Cambridge, England), Victoza
• Small molecule drugs are chemically synthesized (Novo Nordisk; Bagsvaerd, Denmark), and Trulicity
which means made by a series of chemical reactions (Eli Lilly; Indianapolis, Indiana). These peptide drugs
in the lab. They are typically taken as a pill or capsule. work by interacting with a receptor on the surface of
The pill or capsule dissolves in the gastrointestinal pancreatic beta cells and stimulate the release of insulin
tract and the active ingredient is easily absorbed for diabetes.
into the bloodstream via the intestinal wall. The
molecules that make up these drugs are so tiny IN THE RHYTHM
they are able to penetrate cell membranes and get Rhythm Pharmaceuticals (Boston, MA) is conducting
inside of cells.In contrast, large molecule drugs — Phase III clinical studies of their anti-obesity peptide
protein-based therapeutics known as biologics — are drug setmalanotide. Designated as a breakthrough
made by living cells. They must be administered therapy by the FDA, early clinical trial results in rare
via injection because they will be destroyed by genetic forms of obesity were promising, helping to
digestive enzymes in the gastrointestinal tract if attract $41 million from key investors including Pfizer
given orally. Their large size, anywhere from 50 to Venture Investments and Third Rock Ventures to fund
1,000 times larger than a typical small molecule Phase III.
drug, makes it impossible for them to penetrate
Setmalanotide works by activating the melanocortin-4
cells. On the flip side, large molecules are highly
receptor (MC4R), a receptor present on the surface of
specific for their target — typically a cell-surface
cells in the hypothalamus of the brain, a region involved
receptor on the outside of the cell.The FDA defines
in regulating both appetite and satiety. Mutations in
a peptide therapeutic as a chain of amino acids (the
MC4R that result in reduced activation are the most
building blocks of proteins) containing 40 amino
common genetic cause of obesity, and account for
acids or less, and regulates them as small molecules.
approximately 6-8 percent of obesity cases.
Peptide therapeutics are similar to small molecule
drugs in that they can be synthesized in the lab
KALOS FIGHTS CANCER
using a peptide synthesis machine — a machine
that links amino acids together in a specified order. Kalos Therapeutics (San Diego, CA) has a peptide drug
Peptides also share key characteristics with large in development based on a straightforward observation:

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despite constant activity, heart muscles don’t get bigger, cancer cells but is inactivated in a range of malignancies.
and cancers of the heart are extremely rare. At least ALRN-6924 is in Phase II clinical studies for lymphoma.
part of the reason for this is a peptide known as atrial The company is pursuing the development of stapled
natriuretic peptide (ANP), which is produced in the peptides in a range of therapeutic areas, including
heart. It helps to control cell growth and division, making inflammation and endocrine and metabolic diseases.
sure that the heart doesn’t get too big for the chest. Bicycle Therapeutics also uses chemically-linked
Since cancer is caused by out-of-control cell growth and peptides to increase stability, target interaction, and
division, a connection was made: perhaps these peptides penetrate cells. Their peptides are formed — using a
could play a role in controlling tumor cell growth. chemical linker — into the shape of a bicycle.
Kalos Therapeutics has identified a portion of ANP and Bicycle’s lead candidate, BT1718, is a “bicycle drug
is synthesizing and testing it as a potential anti-cancer conjugate” — a bicyclic peptide with a toxic drug
agent. Dubbed KTH-22, the agent is cytostatic, meaning attached. The peptide targets a protein called
it halts the growth and division of cancer cells, but “membrane type 1 matrix metalloproteinase” (MT1-
does not directly kill them as a cytotoxic (toxic to cells) MMP) which is overexpressed in many tumors. BT1718
agent would. KTH-22 is in preclinical research, with data delivers its toxic payload to tumors overexpressing
supporting its use in the treatment of pancreatic and MT1-MMP. BT1718 is now in Phase I clinical studies for
ovarian cancers. solid tumors.

STAPLES AND BICYCLES

Most peptide therapeutics do not penetrate cell
membranes. Designing peptides that could enter cells
would truly endow them with the best characteristics
of both large and small molecule therapies. Aileron
Therapeutics (Cambridge, MA) and Bicycle Therapeutics
(Cambridge, U.K.) are aiming to do just that.
Aileron Therapeutics is developing “stapled peptides.”
These peptides are synthesized according to an
optimized amino acid sequence. Next, a chemical linker
is used to connect two amino acids within the chain, Already capable of affecting a range of therapeutic
creating a folded or “stapled” version of the peptide. targets with high specificity, continued innovations in
These stapled peptides still recognize their target peptide design and delivery should make this class of
protein, are more stable, and better able to penetrate drugs an important player.
cell membranes than the linear versions.
Aileron’s leading stapled peptide candidate, ALRN-
6924, activates p53, a protein that triggers cell death in

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EPIGENETICS: WRITING, READING, & ERASING Life Science Training from Industry Experts

FOUNDATIONS OF EPIGENETICS (CH3) group to cytosine (C) nucleotides. The end result:
methylation reduces or even blocks gene expression.
Genetic mutations — changes in the order of the A, C,
G, and T nucleotide bases that make up a gene — have A second type of modification is called acetylation —
been the primary focus of cancer researchers over the addition of an acetyl group (CH3CO) to the histones.
the last several decades. By sussing out mutations Acetylation loosens the association of the DNA with
involved in regulating cell growth and division, scientists the histones, making the DNA more accessible to the
better understand the molecular range of different enzymes used in gene expression, ultimately increasing
cancers and consequently develop more targeted and protein production.
effective therapeutics.
In recent years, another type of genetic variation has
captured the attention of researchers: epigenetic
modifications. Best characterized in cancer, epigenetic
changes are also thought to play a role in a range
of other diseases, including autoimmune disease,
cardiovascular disorders, diabetes, neurodegenerative
disorders such as Alzheimer’s disease, and potentially
even male infertility. In this WEEKLY, we’ll tell the
epigenetics story and discuss how it’s being used to
develop new treatments.
Deacetylation — the removal of an acetyl group —
TERM OF THE WEEK: EPIGENETICS increases the association or “grip” of the DNA around the
histones. Deacetylation makes the DNA less accessible
Epigenetic modifications are changes to DNA that do
to enzymes used in gene expression, thereby decreasing
not alter the actual gene sequence; they are chemical
the production of proteins.
modifications to the DNA itself. These changes typically
affect gene expression, or how often the gene is read by
ADDING IT ALL UP
the cell. Epigenetic modification can occur either directly
to the nucleotide bases themselves (A, C, G, or T) or to Epigenetic modification is a normal part of development.
the histones, which are small proteins that package and This is in part why different genes are expressed in the
order DNA. heart than, say, the liver — the two different tissue types
contain the same genome, but tissue-specific differences
in epigenetic modification lead to differences in gene
expression in the two tissues.
Problems may arise, however, if variations in epigenetic
modifications result in changes to gene expression.
If a cell or tissue type begins to make too much of a
protein that activates cell growth, for example, the cell
could begin to divide too often — potentially leading to
cancer. Alternatively, a cell could begin to make less of
a protective protein, for example, a “tumor suppressor”
protein (a protein that deactivates cell division), again
One of the most common types of epigenetic potentially leading to cancer.
modification is methylation — the addition of a methyl

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Epigenetic medicine seeks to identify disease-associated DNA-methyltransferase (DNMT) inhibitors: DNMTs
differences in epigenetic modifications, and to develop are writers — they add methyl groups to DNA,
drugs that restore the epigenome to that of healthy cells. resulting in decreased expression of associated genes.
Two DNMT inhibitors have been approved by the FDA:
BREAKING IT DOWN Vidaza (Celgene) and Dacogen (Otsuka; Tokyo, Japan).
Epigenetic drugs are small molecule drugs that target Both drugs are used to treat myelodysplastic syndrome
epigenetic regulators, or proteins that write, read, or and acute myeloid leukemia.
erase epigenetic modifications.
• Histone-methyltransferase (EZH2) inhibitors:
• Writers are the enzymes that make the chemical EZH2s are also writers — these enzymes transfer
modifications — methylation or acetylation methyl groups to histone proteins. One EZH2
as described above — to DNA molecules or is associated with overactivity in a number of
histone proteins. different cancers. There are no EZH2 inhibitors
• Erasers are enzymes that remove these currently approved, but several are in development,
chemical groups. including Constellation Pharmaceuticals’
(Cambridge, MA) CPI-1205 in Phase I for advanced
• Readers are the proteins that detect and respond to
B-cell lymphomas, and Epizyme’s (Cambridge, MA)
these modifications, causing the DNA to be more or
tazemetostat, currently in Phase II for non-Hodgkin
less tightly wrapped around the histone protein.
lymphoma, certain genetically-defined solid tumors,
Any of these proteins could be inhibited or activated to and mesothelioma.
affect changes in epigenetic modifications.
THE NEW CLASS: READERS
OLD SCHOOL: WRITING AND ERASING A class of proteins called “Bromodomain and Extra
The disease that has been best classified in terms of Terminal motif” (BET)proteins are reader proteins. They
epigenetic variations is cancer. Currently, there are five recognize and bind to specific patterns of acetylation on
epigenetic drugs on the market to treat cancer, and histone proteins. Upon binding, they recruit additional
more in development. Those on the market fall into two proteins that regulate gene activity. Irregularities in
categories - erasers and writers: histone acetylation, then, may send the wrong message
• Histone deacetylase (HDAC) inhibitors: HDACs to a BET protein. By inhibiting the interaction between
are erasers — they remove acetyl groups from BET proteins and histone proteins, researchers have
histone proteins, resulting in increased expression of found that they can prevent incorrect messages from
associated genes. Three HDAC inhibitors have been being received by the BET proteins. Currently, there
approved by the FDA: Zolinza (Merck; Kenilworth, NJ) are no BET inhibitors (BBI) approved, but several are in
and Istodax (Celgene; Summit, NJ) treat cutaneous clinical development. The farthest along is Resverlogix’s
T-cell lymphoma, and Farydak (Novartis; Basel, (Calgary, Canada) apabetalone, which is in Phase III
Switzerland) treats multiple myeloma. HDAC testing for atherosclerosis and associated cardiovascular
inhibitors in development include those in the disease. Additional BBIs in clinical development are
table below: shown in the table below:

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Epigenetics promises to change the way we look at the work in progress. As that work develops, researchers will
human genome. Scientists have made great strides in undoubtedly uncover new drug targets and approaches
understanding how epigenetic modifications contribute to disease management. Stay tuned!
to both health and disease; however, a complete
understanding of these modifications is still very much a

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BREATH BIOPSIES Life Science Training from Industry Experts

A BREATH OF FRESH AIR patients based on metabolic changes – many of which

FOR DIAGNOSTICS appear before other symptoms.

In the scary world of serious illness, early detection is Science has long known that cancer cells have irregular
crucial. The sooner someone knows he has cancer or metabolisms. For example, they consume glucose at a
rheumatoid arthritis, for example, the sooner they can rate as much as two hundred times greater than that of
get moving with treatment. Early detection means a healthy counterparts. Malignant cells also use anaerobic
greater likelihood of a good outcome, saved time and (without oxygen) metabolic pathways, rather than the
money and maybe even less heartache. aerobic – with oxygen –- pathways preferred by healthy
cells. Cancer’s altered cellular rhythm manufactures
Sadly, diagnosing disease can be grueling, painful, and
different metabolic by-products, some of which can
time consuming. Undoubtedly, patients know this best.
be detected as VOCs. Other diseases produce altered
However, medical researchers know it too. Some of
VOCs too, including diabetes, liver impairment, kidney
them are working to improve the process. Maybe you’ve
disease, asthma, cystic fibrosis, rheumatoid arthritis,
heard of liquid biopsies – the detection of cancer in a
atherosclerosis, and tuberculosis.
drop of blood or a urine sample rather than instead
of a traditional biopsy. If you think that’s amazing, These metabolic changes that come with illness underpin
what about a breath biopsy? Think of it as a cancer the whole notion of the disease breathalyzer.
breathalyzer. This WEEKLY takes a closer look at this
innovative approach to detecting illness.
BREATHING LIFE INTO
VOC DIAGNOSTICS
TERM OF THE WEEK: VOLATILE Owlstone Medical (Cambridge, U.K.) has begun to
ORGANIC COMPOUND develop a diagnostic based on VOCs with their ReCIVA
Here’s a possibly new perspective on that peanut butter Breath Sampler. During testing, a patient inhales
sandwich or banana you may have just munched. It may normally from a standardized air supply, to make sure
have tasted like food but really it’s fuel. Specifically, it’s that the source of air doesn’t impact the results. The air
energy that our bodies use to power cellular function is then exhaled into a breath biopsy cartridge, which
courtesy of a series of chemical reactions. The whole captures the patient’s VOCs. The cartridge is then sent
miraculous affair, cellular metabolism, produces to a lab for analysis with Owlstone’s proprietary Field
and ultimately releases organic (carbon-containing) Asymmetric Ion Mobility Spectrometry (FAIMS).
compounds into our blood. When the blood passes
through our lungs, some compounds are exchanged into
WHAT’S NOT TO LOVE?
the lungs, and then exhaled. Once we breathe the stuff Breath biopsies possess a unique, obvious advantage
out, scientists call them volatile organic compounds over other diagnostic methods. No pain, no discomfort.
(VOCs). Our breath contains more than a thousand They don’t even require a finger prick! The test is
VOCs, or metabolites, as they are also called. also extremely sensitive – detecting even miniscule
concentrations of suspicious VOCs. That’s because it
THE DISRUPTIVE RHYTHM OF DISEASE takes our entire blood supply only about a minute to
Disease affects our cellular metabolism, changing circulate throughout the body. Yup, sixty seconds. The
which VOCs it produces. The unique VOC “fingerprint” sampling period for breath biopsies is typically ten
of different maladies may help researchers identify minutes. That allows all of our blood to get sampled
several times, making the discovery of a questionable
VOC extremely likely.

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IN THE WIND including bladder, breast, head and neck, kidney,
esophageal, pancreas, and prostate.
Unfortunately, no breath biopsies are on the market yet.
However, Owlstone is already conducting several clinical COCKTAIL FODDER: CATCHING
trials to demonstrate its validity: A WHIFF OF CANCER
• LuCID (Lung Cancer Indicator Detection): In Whether you consider dogs best friend material or
partnership with the U.K. National Health Service, merely tolerate them, take one minute to appreciate
this trial has identified unique VOCs in lung cancer their amazing snouts. Canine noses contain 220 million
patient’s exhalations. Phase II aims to validate the scent receptors. Puny little human noses only boast
detection of these biomarkers in clinical practice. about five million. Their superior scent-detection enables
• STRATA (Stratification of Asthma Treatment by these super-sniffers to find lost people, explosives and
Breath Analysis): This study is examining how VOCs contraband of all sorts. Scientists have also discovered
may help select the best asthma treatment. that dogs can sniff out some kinds of cancer. That’s
• InTERCEPT: This study focuses on identifying and because VOCs are present in all of our emissions! Breath,
validating VOCs to detect colorectal cancer. sweat, saliva and, well, you know. Fido definitely knows
that cancer stinks!
• PAN (PAN-cancer early detection): In partnership
with Cancer Research UK, this trial aims to identify
VOCs associated with many different cancers,

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FOUNTAINS OF HEALTHY OLD AGE? Life Science Training from Industry Experts

FOUNTAINS OF HEALTHY OLD AGE? of Type 2 diabetics strongly suggests that people who
take the drug live longer than subjects of the same
Plenty of famous people have uttered (or supposedly
age who don’t. Researchers from the Albert Einstein
uttered) lots of pithy comments about aging. A couple
College of Medicine’s Institute for Aging Research
gems: “Age is mind over matter; if you don’t mind, it don’t
(Bronx, NY) have begun clinical testing metformin to see
matter,” said Satchel Paige. “We are always the same age
whether it reduces the overall incidence of age-related
inside,” Gertrude Stein wrote — she lived in Paris; how
diseases. The premise that growing old itself deserves
could she help but feel amazing?
intervention represents a paradigm shift for the FDA and
Famous or no, longevity fascinates us, from legends for our culture, opening the door to more studies aimed
about fountains of youth to today’s theories about at improving health span.
restricting calories and quaffing red wine. As our
understanding of aging deepens, more companies SUPPRESSING THE IMMUNE
are working to clinically validate and commercialize SYSTEM FOR LONGER LIFE?
treatments that extend life span and more importantly, As metformin shows, off-label use of approved drugs
extend health span, or the period of a person’s life sometimes yields promising results. For example,
during which they are generally healthy and free from multiple studies of the immunosuppressant rapamycin,
serious or chronic illness. used to prevent rejection in transplant patients, show
Healthy aging depends on a host of factors — genetics, that the drug lengthens lifespan in mice.
lifestyle, socioeconomic status, environment, nutrition. Rapamycin inhibits mTORC1, an enzyme that helps cells
To move ahead, the industry needs to tease out these detect whether they have the proper levels of critical
complications and integrate them into a holistic picture. nutrients and to respond appropriately. Defects in
This week and next, we’ll meet some of the players mTORC1 signaling are linked to a range of age-associated
working hard to do that. diseases including Type 2 diabetes, Alzheimer’s, and
rheumatoid arthritis.
SHIFTING THE PARADIGM
Rapamycin completely inhibits mTORC1 and somewhat
Many clinical trials have focused on conditions
inhibits the related enzyme mTORC2. Complete
associated with old age, such as dementia, cancer, and
inhibition of mTORC1 makes rapamycin unsafe for long-
cardiovascular disease, rather than growing old itself.
term use. Enter Navitor Pharmaceuticals (Cambridge,
Now researchers are beginning to parse the underlying
MA), which seeks to develop “selective modulators” of
molecular pathways associated with aging. The next step
mTORC2. Instead of completely blocking the enzyme,
is to identify and develop drugs that influence those
the new compounds would selectively modify mTORC1.
pathways. If successful, we may get at the root cause of
ResTORbio (Boston, MA) is also working on selective
diseases associated with aging, rather than tackling one
mTORC1 inhibitors.
infirmity at a time.
BLOODY SUCCESS!
OFF-LABEL & ANTI-AGING
One extremely intriguing area of longevity research
Some of the most promising leads come from exploring
sounds like it was inspired by Dracula or the Twilight
off-label uses of current medicines. Researchers have
movies saga. Tony Wyss-Coray and his research
discovered that the diabetes drug metformin may have
group at Stanford University (Palo Alto, CA) doubly
anti-aging benefits. The drug significantly extended
demonstrated that old mice exposed to the blood of
lifespan in animal models.
young mice experienced increased neuron growth.
Research also indicates that metformin can reduce the
First, parabiosis. The word means “living beside.” In
risk of cancer and dementia. Moreover, a large study
Wyss-Coray’s study, researchers surgically joined pairs

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of mice to share one circulatory system. This technique ZEROING IN ON THE X FACTOR
revealed that exposure to young blood enabled geezer
For the longer term, the next step may be isolating the
mice to repair damaged liver and muscle tissue, probably
exact beneficial protein or proteins which promise to be
by activating stem cells.
safer and more effective than whole-plasma treatments.
Wyss-Coray also showed that simply injecting the Last April, Alkahest began Phase II clinical testing
“mature” mice with plasma, the protein-y part of blood, of GRF6019, a proprietary elixir of plasma proteins
from young mice achieved the same effect —– good researchers believe cause rejuvenating effects. This
news for larger, bipedal mammals like us. study examines the results of GRF6019 on Alzheimer’s
A study involving human umbilical cord blood is patients. There’s also potentially good news for people
perhaps even more impressive. This blood also showed with Parkinson’s disease. Alkahest has received a grant
rejuvenating effects on geriatric mice, specifically in from the Michael J. Fox Foundation (NY, New York) to
learning and memory. begin testing the product in those patients.
To propel such findings down the biopharma pipeline, Join us next week as we continue our overview of
Wyss-Coray founded Alkahest (Menlo Park, CA). Last the science behind longevity research by examining
fall, the company unveiled the outcome of a Phase I telomeres, cellular senescence, and genomics research
clinical study on the effects of infusing plasma from as part of the quest to extend healthy lifespan.
young donors (under 30) into Alzheimer’s patients over
50. According to a press release, the older patients’
functional activity improved in statistically-significant
ways and they tolerated the treatment well.

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HACKING THE FOUNTAIN OF YOUTH Life Science Training from Industry Experts

HACKING THE FOUNTAIN OF YOUTH SENESCENCE

Last week, we began to delve into longevity research This beautiful word refers to an unpleasant reality. Our
— the area of biotech which seeks to understand ever-shortening telomeres doom our cells. This is cellular
the biomolecular changes that occur with aging and senescence, the state in which cells can no longer divide.
possibly underlie many of the dubious gifts of old age: Senescent cells are found throughout our bodies. They
heart disease, Alzheimer’s and more. The work aims to tend to collect in areas that wear out such as eyes and
lengthen life span of course, but more importantly to joints. Senescent cells secrete signaling molecules that
extend health span — the years in which the average increase inflammation, which drives several aging-
person functions independently at a high level, both related diseases including cancer, cardiovascular disease,
physically and mentally. This week, we’ll explore osteoarthritis, and Alzheimer’s.
research surrounding telomeres, cellular senescence, Studies in mice show that clearing senescent cells
and genomics. lengthened their lives by an average of 20 percent. It also
delayed organ deterioration in healthy mice. This success
TELOMERES: THE LONG has inspired the development of “senolytics,” a new class
& THE SHORT OF IT of drugs that target and destroy senescent cells. Unity
Scientists have long known that the length of telomeres Biotechnology’s (Brisbane, CA) lead candidate, UBX0101,
has implications on aging. First though, what are is in Phase 1 clinical testing for osteoarthritis. Cleara
telomeres? Sort of simply put, they’re the bits of DNA at Biotech(Utrecht, Netherlands) and the Mayo Clinic
the end of all chromosomes with a repeating sequence (Rochester, MN) are also exploring senolytic treatments
of six nucleotides: TTAGGG. This structure protects for cancer and other aging-related diseases.
chromosomes from losing important information
during replication. IT’S A BIRD, IT’S A PLANE, NO!
Old telomeres get shorter, because our adult cells lack IT’S SUPERCENTENARIAN!
an active telomerase enzyme. The enzyme ensures that Lifestyle certainly influences health span. For most, a
the entire telomere gets copied. Telomere length may healthy weight, not smoking, and exercise should add a
predict life span — shorter telomeres correspond to few years of mental and physical fitness. However, the
shorter lifespan. oldest recorded living human, Jeanne Calment, died in
Scientists at Stanford University and Houston 1997 at age 122. Her demise came five years after kicking
Methodist Research Institute have induced cells a one-hundred-year-old smoking habit. A century of
to produce telomerase by delivering a modified death sticks! Other supercentenarians (people older than
telomerase-coding mRNA molecule to human muscle 110) have boozed it up and enjoyed copious amounts of
cells grown in the lab. The mRNA temporarily provides sugar (Ms. Calment supposedly ate over two pounds of
the cells with instructions for making telomerase; chocolate per week — sacre’ bleu!).
within a few days these protein-making instructions are Most supercentenarians studied have somehow
degraded and the enzyme is no longer produced. This remained mentally sharp and free from serious
temporary expression is key — unchecked telomere chronic conditions, suggesting that protective genetic
extension can cause cells to turn cancerous. This factors play a role. Just what those factors are is the
temporary induction of telomerase did indeed promote million-dollar question. Answering it poses a challenge,
a significant lengthening of the telomeres. For now, the especially due to the small pool of super-ancients. A few
method has only been used in the lab to increase the life groups are trying. The New England Centenarian Study
span of cells, but it may have future clinical potential as and the New England Supercentenarian Study, both
an anti-aging treatment in humans. at Boston University School of Medicine, have been
studying elderly and very elderly genomes for genetic

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traits that set them apart. From the studies’ website, GENE THERAPY
preliminary findings include:
Any discussion of longevity science needs to include the
• The genetic influence increases with older ages possibility of cutting-edge gene therapies. Legendary
of survival. genomics pioneer and Harvard Medical School
• Especially in people over 105, diseases of aging professor George Church founded Rejuvenate Bio
typically don’t appear until the very end of their lives, (Cambridge, MA) to test gene therapy that extends
if at all. dogs’ lives. Their ultimate goal: helping dogs’ best
• This genetic influence probably involves many friends live longer and better. Rejuvenate Bio is looking
variants with individually modest effects; however, into a treatment that turns on genes that arrest the
as a group, their effect is strong. progression of heart failure. The scientific literature also
mentions genes that increase muscle mass and that
• Excluding rare exceptions, centenarians carry as
activate telomeres as potentially relevant to the study
many disease-associated genes as the rest of us. This
of longevity.
suggests that their genetic advantage results from
gene variants that slow aging and decrease risk for Longevity and health span research is an already large
aging-related disease–protective genes. and burgeoning area of biotech. This WEEKLY covers
some of the most interesting highlights. For a more
This research aims to identify those protective genes
comprehensive look, visit this 800 page (!) report. If even
and develop interventions to improve the health
some of these companies succeed, we can look forward
span of those of us who endure the typical insults
to thinking clearly and moving well into our tenth
of Father Time. Other explorations of the genetics
decade. L’Chaim!
of aging include the “Wellderly” project at Scripps
Translational Science Institute (La Jolla, CA) and the
Betterhumans Supercenturian Research Study. The
latter is a collaboration of BIOAGE (Richmond, CA), the
Methuselah Foundation (Springfield, VA), Veritas
(Danvers, MA); and the Albert Einstein College of
Medicine’s Institute for Aging Research (New York
City, NY).

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ATTACK OF THE TREGS!! Life Science Training from Industry Experts

ATTACK OF THE TREGS!! encounter the activating protein, they release anti-
inflammatory cytokines.
No matter what it sounds like, this subset of helper
T-cells aren’t invaders from another planet. Nope, Tregs, TxCell (Valbonne, France) has CAR-Tregs in preclinical
or regulatory T-cells, come from New Jersey (don’t development for graft-versus-host disease and
say it!) and elsewhere. They suppress the immune autoimmune/inflammatory disorders including multiple
system. These newcomers to the drug discovery and sclerosis, inflammatory bowel disease, rheumatoid
development scene intrigue the men and women seeking arthritis, and inflammatory skin disease.
new approaches to diabetes, cancer, and autoimmune
disorders. This WEEKLY examines Tregs — their function
TREGS VS. TYPE 1 DIABETES
and what they can do for us. Caladrius Biosciences (Basking Ridge, NJ) is conducting
Phase II clinical studies of a Treg-based treatment for
Tregs are a kind of T-cell that prevents other immune
Type 1 diabetes. This autoimmune disorder turns a
cells from attacking the body’s tissue and other harmless
person’s natural defenses against her or his insulin-
cellular “stuff” such as food or friendly bacteria. Like
producing pancreatic cells. The new treatment stems
other T-cells, Tregs switch on when their receptors
from the fact that in most Type 1 diabetics, up to
recognize a particular activating protein, or antigen.
twenty percent of their insulin-producing cells are
The specific portion of the antigen the Treg receptor
intact at diagnosis. If a treatment can protect those
responds to is a “Tregitope.” Scientists haven’t totally
cells and restore their function, it may halt the disease’s
figured how this suppression takes place. They do know
progressive destruction. This, in turn, would make
at least some of the effect comes from the release of
controlling patients’ blood glucose levels easier and
anti-inflammatory signaling molecules —cytokines.
ultimately reduce the risk of long-term complications.
WHEN TREGS GO WRONG Caladrius’ Type 1 diabetes product, CLBSO3, consists of
Defective or too few Tregs can cause the attack cells of an individual’s own Treg cells that health care providers
the immune system — think killer T-cells, macrophages modify and readminister.
— to go into overdrive, causing severe inflammatory
disease. Too many Tregs aren’t good either. Tregs are
TREGITOPES
present in abnormally high numbers in different types of Researchers at EpiVax (Providence, RI) hope to bypass
solid tumors. This excess of Tregs makes it harder for the cell therapy altogether by relying on Tregitopes — the
immune system to recognize and fight off cancer cells. short amino acid sequences that activate subsets of
Tregs. The idea is to give patients Tregitopes with an
AMPING UP TREGS antigenic protein, in hopes of triggering antigen-specific
Like CAR-Ts, CAR-Tregs are engineered T-cells. However, Tregs within the patient. EpiVax is now conducting
instead of unleashing tumor-killing power, the goal is to preclinical studies for Type 1 diabetes and allergies. The
unleash the calming power of Tregs by activating them to company anticipates their approach will help prevent
release anti-inflammatory cytokines. immune reactions to interventions like therapeutic
proteins, organ transplants, and gene therapies.
To make CAR-Tregs, researchers isolate Tregs from a
patient’s blood and then engineer them to contain a SWITCHING ON THE TREGS
special receptor. Once reinfused into the patient, the
Nektar Therapeutics (South San Francisco, CA), in
new receptor directs the Tregs to recognize a protein
partnership with Eli Lilly (Indianapolis, IN), is developing
from inflamed areas of a patient suffering from an
NKTR-358. This biologic drug activates Tregs by
autoimmune or inflammatory disorder. When they
interacting with an activation receptor on the surface of

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Tregs. NKTR-358 is currently in Phase I clinical trials for • Bristol Myers Squibb (New York, NY) is developing
systemic lupus. small molecule Treg inhibitors.

TREGS VS. CANCER TREGS VS. OBESITY & ITS CRONIES

An overabundance of Tregs contributes to the way the In recent years, researchers have realized that
tumor microenvironment subdues the immune system. inflammation, obesity, and Type 2 diabetes are linked.
This could mean that suppressing Tregs themselves may This relationship points to the possibility that Tregs may
someday treat cancer. Here are some possibilities: be able to modulate obesity-associated inflammation.
• Tizona Therapeutics (South San Francisco, CA) is Still years away from the clinic, scientists at the Mathis
developing a monoclonal antibody therapeutic that Lab at Harvard are investigating.
targets Tregs, with the goal of removing them from The immune system is a powerful tool that has already
the tumor’s “neighborhood.” been enlisted as a potent weapon against cancer. As
• AbbVie (North Chicago, IL), in partnership with researchers develop more ways to harness our bodies’
Argenx (Belgium), is working on an inhibitor of natural defenses, the biotech industry may revolutionize
GARP, a Treg surface protein that enables anti- other areas of medicine en route to optimizing the
inflammatory signaling molecules to be produced. balance between overactive and under-active immunity.
Hampering GARP promises to reign in Tregs’
immune-suppressing power.

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AUTOPHAGY: THE INCREDIBLE, EDIBLE CELL? Life Science Training from Industry Experts

AUTOPHAGY: THE INCREDIBLE, Autophagy can also be also induced by cellular stress.
EDIBLE CELL? Sometimes the stress arises from nutrient deficiencies.
Sometimes it comes from a lack of oxygen (ischemic
True, weird fact: our cells are cannibals. Right now, these
injury), infection, or the presence of damaged proteins
itty-bitty sacs of vital fluids inside your body are eating
or organelles.
themselves. This phenomenon is called autophagy, or
“self-eating.” CELLULAR CANNIBALS & THE BRAIN
AUTOPHAGY IS AWESOME Scientists suspect that autophagy protects our brains by
flushing out toxic proteins that accumulate in disorders
This seemingly odd phenomenon is old news to
like Huntington’s disease, Alzheimer’s disease and
biologists. They already know that autophagy is really
Parkinson’s disease.
about cellular cleanup and recycling, not cannibalism.
It’s a highly regulated process by which cells disassemble Defects in autophagy have been associated with
unnecessary or broken bits into their component the aggregation of mutant huntingtin proteins in
building blocks. Our bodies then use what used to be Huntington’s disease and with the buildup of amyloid-
junk to make new parts. beta plaques in Alzheimer’s disease. And in Parkinson’s
disease (PD), defects in mitophagy are commonly
Alas, like many bodily functions, autophagy sometimes
associated with inherited types of PD. “Mitophagy” refers
run amok. Scientists have implicated problems with
to autophagy directed at mitochondria – an important
the process in a range of health issues including
process for keeping these energy-producing factories
neurodegeneration, cardiac disorders and cancer. That’s
healthy enough to fuel neuron function.
bad news for those of us with bodies.
The good news is that biotech firms have noticed that CANCER’S FOE; CANCER’S FRIEND
out-of-whack autophagy can wreak havoc on our bodies. Autophagy is a double-edged sword when it comes to
Many are now working on interventions that activate or cancer. It helps suppress tumor formation thanks to its
inhibit autophagy. The efforts may yield new approaches ability to protect cells from stress. However, if a tumor
to disease and illness. does end up developing, autophagy may end up feeding
the tumor’s cells, instead of the body’s. In fact, most
CELLULAR RECYCLING & REUSE tumor cells display increased rates of autophagy.
Autophagy occurs continuously at low levels in most
cells. Here’s what happens: damaged or defective CARDIAC HEALTH
cell parts such as proteins, lipids, and even entire Heart attack blocks oxygen flow to the heart, resulting
substructures (organelles) such as mitochondria, the in tissue death. Recent studies show that increased
energy-providing “powerhouses” of the cell, are enclosed autophagy can help minimize this damage. Abnormal
in lipid vesicles – bubble-like containers composed of the autophagy has also been associated with other problems
same material as the cell membrane. The loaded vesicles including muscular problems, liver disease, and
then fuse with an organelle, called a lysosome, which inflammatory disorders.
has an acidic pH and contains digestive enzymes. The
lysosome works somewhat like a stomach—its caustic IN THE PIPELINE
“juice” breaks down cellular junk into its constituent bits.
Biotech companies have begun to look at ways of
The “cellular stomach” then releases these back into the
harnessing the powers of autophagy for good. Let’s take
cell to form new components. Autophagy thus serves
a look at some of the possibilities:
two purposes: “cleaning out” old or defective parts, and
providing new building blocks. • Casma Therapeutics (Cambridge, MA) was
launched earlier this year to find ways to boost

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 autophagy to treat a range of diseases. The function of lysosomes. Obstructing their function
company is now conducting clinical trials in muscle decreases autophagy. Relieving the inhibition should
disorders, liver disease, inflammatory disorders, increase autophagy.In 2015, a small Phase I study of
and neurodegeneration. Tasigna in Parkinson’s patients showed significant
• PhoreMost (Cambridge, U.K.), in collaboration improvement in these patient’s symptoms. A new,
with the University of Cambridge, is conducting Phase II study will demonstrate whether these
preclinical studies to identify drug targets to increase promising results hold true for a larger group
autophagy in neurodegenerative disorders. of patients.

• Phase II clinical studies are currently being Autophagy promises to be an important mechanism for
conducted on Novartis’ (Basel, Switzerland) cancer a range of different therapeutic areas. These are still
drug Tasigna as a treatment for Parkinson’s and early days in translating these findings from the lab to
Alzheimer’s disease. Originally approved to treat the clinic, but stay tuned – this complex story has only
chronic myelogenous leukemia, the drug works by just begun.
inhibiting an enzyme that promotes cell division.
Tasigna also blocks a protein that interferes with the

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IMMUNE CHECKPOINT INHIBITORS Life Science Training from Industry Experts

A KILLER(-T) NOBEL PRIZE IN MEDICINE suppressing proteins, making it almost impossible for
T-cells to mount an attack.
On October 1, James Allison, now at the M.D.
Anderson Cancer Center in Houston, and Tasuku PRIZE-WINNING RESEARCH
Honjo, now at Kyoto University, won the 2018 Nobel
In the mid-1990s, Dr. Allison hypothesized that inhibiting
Prize in Physiology or Medicine. The two scientists
the CTLA4 protein on T-cells would enable those T-cells
discovered the basis for today’s hugely successful
to become more fully activated in their attacks on
immune checkpoint inhibitor therapies. Let’s look at
malignant cells. Allison then developed a monoclonal
their discoveries and how they have revolutionized
antibody to bind CTLA4, blocking its ability to put the
cancer treatment.
brakes on our immune system. Injecting the antibody
KEEPING THE IMMUNE into mice eradicated established tumors, providing
SYSTEM IN CHECK the rationale for future checkpoint inhibitor therapies.
Dr. Honjo performed similar work with the PD1
In the early 1990s, Allison and Honjo independently
checkpoint protein.
discovered two proteins that serve similar functions in
our immune system. They both shut down killer T-cells,
a type of white blood cell. We want our killer-Ts working
hard to defend us against possible threats. But we also
need to make sure they don’t attack our own healthy
tissue. So, our immune systems need a mechanism that
“checks” a dangerous overactivation of T-cells.
Both proteins, CTLA4 and PD1, are displayed on the
surface of T-cells. When these proteins encounter
proteins on the surface of our own, healthy cells, their
deactivation power switches on. This biological safety
feature prevents overactive T-cells from attacking our
own organs and tissues. Thus, CTLA4 and PD1 are known FROM MICE TO PEOPLE
as immune system checkpoints.
The experiments in mice were the first step towards
groundbreaking new cancer drugs. To translate this
CRAFTY CANCER
basic research into the clinic, Princeton-based biotech
The medical community has long believed that the company Medarax (later acquired by Bristol-Myers
immune system plays an important role in eliminating Squibb) stepped in to develop antibody inhibitors of
cancerous cells as they emerge and before they develop checkpoint proteins that would be safe for humans. The
into a significant problem. It does this by recognizing first of these, ipilumumab (later named Yervoy), targeted
“tumor antigens” or proteins on the surface of overly- CTLA4 and began clinical trials in 2000. Yervoy was
mutated cells that T-cells and other immune cells will the first antibody to target the immune system in the
react to as threats. treatment of cancer rather than targeting the malignancy
However, cancer cells begin their lives as normal cells. itself. In 2011, Yervoy was approved to treat melanoma.
So they also display proteins that instruct the immune Since 2011, the FDA has approved several other
system to ignore them or, in other words, activate checkpoint inhibitors, including those for PD1, a
immune system checkpoints. Some tumor cells have checkpoint protein on the surface of T-cells, and PD-L1, a
actually evolved to make extra copies of these immune- protein on the surface of cancer cells that activates PD1
and is overexpressed on some types of tumors.

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The table below lists checkpoint inhibitors that have • They promise to fight a range of cancers, unlike most
subsequently received approval: antibody therapies, which target one specific type
of cancer. However, their efficacy will vary among
different subcategories of patients.
Researchers are working on identifying the next round
of immune system checkpoint inhibitors by deciphering
proteins such as LAG-3, TIM-3, TIGIT, VISTA.

FINDING OUT
Physicians try to identify the patients most likely to
respond to a particular drug. For example, the best
criteria for giving someone a PD-L1 inhibitor drug is if
their bodies overexpress the PD-L1 protein on the tumor
surface. Agilent Technologies (Santa Clara, CA) markets
NEW AND POSSIBLY IMPROVED
a companion diagnostic to identify patients whose
PD-1 or PD-L1 inhibitors may have one crucial tumors overexpress that particular protein. There may
advantage over CTLA-4 inhibitors: fewer of the potential also be other ways to find patients right for checkpoint
autoimmune-like side effects seen in CTLA-4 inhibitor treatment. Early studies indicate that genetic signatures
drugs. PD-1/L1 inhibitors’ decreased side effects may of tumors may help identify who will respond well to
result from the fact that they seem to primarily activate PD-1 inhibitors. The diagnostics company NanoString
T-cells already present in tumors’ tissues, rather than Technologies (Seattle, WA) is developing a genetic
those present in healthy tissues that would be damaged test that predicts who is most likely to benefit from
by a T-cell attack. checkpoint inhibitor therapies. Another possibility lies
in looking at tumor biopsies to see if T-cells are already
LOOKING AHEAD present. If so, it’s likely that switching them on via
Oncologists are excited about these new therapies for checkpoint inhibitors will treat their cancer.
two main reasons: The past few years have yielded a revolution in medicine.
• They are proving to be longer-lasting than other Clinical researchers are finally translating the insights of
treatments, even when compared to the highly a few brilliant immunologists into a clinical reality. As a
effective, targeted monoclonal antibodies. Once new generation of researchers and biotech companies
activated by checkpoint therapy, the immune system continues to push the boundaries of knowledge
can evolve and change with the cancer, unlike and application, expect to see even more exciting
static therapies. revolutions ahead.

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INFLUENZA: SO S’NOT FUNNY Life Science Training from Industry Experts

INFLUENZA: SO S’NOT FUNNY B IS FOR BODY SLAM

Happy Flu Season! A throbbing headache, fever, chills—if The flu virus uses proteins on its surface to dock onto
you’ve ever enjoyed a few days off courtesy of influenza, and invade our lung cells. Once inside, the pathogen
you’ve got good reason to embrace the simple flu shot. highjacks cellular machinery to replicate itself. The
Every year, millions of us roll up our sleeves and take our copies are then released to invade the nearest
medicine. This WEEKLY reviews the ABCs of the flu virus neighbors. This process does damage lung cells
and what you need to know about the vaccine. somewhat. But most symptoms actually stem from
The US Centers for Disease Control (CDC) (Atlanta, the immune response itself. Viral replication attracts
GA) estimates that about 170 million doses of influenza T-cells. To stamp out the threat, these warrior cells do
vaccine will go to doctors’ offices, health departments, what they do best: kill infected cells. The cellular death
and even the corner drugstore to help keep people damages our lungs and kicks mucus production into
flu-free during 2018-19. Drug companies deal with this high gear. Voila—cough, cough hack, sneeze, yuck. All of
high demand by making the vaccines six to nine months this unpleasantness is good though. It means your poor
before flu season starts in October. aching body is trying to get rid of flu filth.
Activated T-cells also release inflammatory cytokines
A IS FOR AWFUL – signaling molecules that further rev up immune
Last year’s flu season was terrible—and not just for the response, sending more white blood cells to attack
young, elderly, and immunocompromised, all of whom the infection. Cytokines also contribute to symptoms.
are especially prey to the illness. The CDC tallied a For example, some affect the region in the brain’s
record number of flu-related hospitalizations. Sadly, the hypothalamus that controls temperature, leading to
season also killed 80,000 people in the US alone. That’s fever and headache. Others get into our muscles. In fact,
the highest number of flu-related deaths since the CDC research indicates that inflammatory cytokines break
began tracking flu mortality in 1976 (CDC). down muscle tissue by switching on muscle-degrading
genes – leading to, you guessed it, muscle aches.
Like all viruses, a flu bug has a simple structure. It
consists of eight strands of RNA – its genetic material - Although our immune response makes us miserable,
surrounded by a protein capsid, all nestled within a lipid we are far better off with than without it. The elderly or
envelope. With influenza, the RNA encodes 11 genes, other immunocompromised flu sufferers experience
each of which produces a different protein. more damage to lung cells than younger, more robust
folk. Unfortunately, that means they are far more likely
to suffer complications, such as secondary infections like
bacterial pneumonia.

NOT ALL BODY SLAMS ARE ALIKE

A few years ago, you may have heard of the H1N1 flu.
What’s up with the letters? “H” refers to the protein
hemagglutinin, which is present on the virus surface.
It facilitates entry into host cells. “N” stands for
neuraminidase, another surface protein. It enables the
release of new viral particles from infected cells.

How does a seemingly inconsequential little microbe Hemagglutinin and neuraminidase both have variations.
wreak such havoc? Irritatingly, most of the illness’s Particular combinations are identified by number. Hence
misery results from our own bodies trying to defend us. H1NI or H3N2, for example. Certain combinations seem

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to make some flu seasons worse. Scientists are still season; however, these predictions are notoriously
trying to figure out why. Theories include the possibility tricky. Regardless though, flu vaccines offer protection.
that it’s more difficult to develop a reliable vaccine They do not, repeat, do not –make people sick, no matter
against certain influenza strains or the fact that some what you may hear otherwise at the water cooler. Some
types of hemagglutinin target more tissues, resulting in people do feel minor symptoms associated with the
nastier symptoms. vaccine, such a slight fever or headache. These actually
mean that the vaccine is kicking in and prodding the
C IS FOR COMMON SENSE: body’s immune response into action.
GET YOUR VACCINE
The best way to avoid the flu is to get vaccinated. The GOOD NEWS ON THE HORIZON
idea behind vaccination is simple. Exposing the immune A number of different biotech companies are working
system to an inactivated virus “trains” it to quickly towards a “universal flu vaccine.” This shot could be
recognize the real thing in the future. given once every five years or so and be more effective
Most vaccines are close to 100% effective. Alas, flu than annual vaccines. Next week, we look at the
vaccines are more typically about 60% effective. This technology behind these vaccines, as well as review
is because the flu mutates considerably from season some new flu drugs in development. In the meantime,
to season. CDC epidemiologists work hard to predict sneeze into your elbow, wash your hands and get that
the most prevalent strain of the virus for the upcoming flu shot!

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UNIVERSAL FLU VACCINE: COMING
SOON TO A PHARMACY NEAR YOU? Life Science Training from Industry Experts

SQUASH THAT BUG

The flu. Need we say more? Why yes. Yes, we do!
Last week, we examined all things flu: we talked
about the influenza virus structure, how the pesky
pathogen makes us feel so lousy, and why the vaccine’s
effectiveness isn’t always up to sniff. Er, snuff. This
WEEKLY delves deeper into the season’s favorite virus.
More specifically, we look at some high-profile efforts to
transform the flu vaccine into the pandemic-crusher of
the WHO’s (World Health Organization) dreams.

The nine epitopes selected are conserved, which

AN ALTERNATE FLU-NIVERSE
means they are present on different HA proteins across
In February 2018, the National Institute of Allergy and different strains. So if the BiondVax vaccine successfully
Infectious Disease (NIAID) unveiled a strategic plan to prompts an immune response, it should protect against
create a vaccine that better protects against multiple different strains of the flu over multiple seasons.
strains of the virus for multiple years. In other words, a
Preliminary data suggests that this mixture of epitopes
universal flu vaccine. Just a few months later, the Gates
will induce not only an antibody response, as most
Foundation (Seattle, WA) announced plans to earmark up
vaccines do, but also a T-cell response. The current flu
to $12 million to support the development of a universal
shot doesn’t. Once activated, T-cells quickly kill virus-
flu vaccine. Let’s check out some of the science behind
infected cells. And to frost the immunity cake, the
these initiatives.
vaccine also produces Memory B- and T-cells that can
quickly respond to infection by a live virus. M-001 is in
THE EPITOPE HOPE
Phase III clinical studies.
Flu prevention today relies on whole-virus vaccines.
Researchers at the Icahn School of Medicine at Mt. Sinai
They deliver an inactivated version of the virus, which
(New York, NY), in collaboration with GlaxoSmithKline
elicits an immune response in the person rubbing her
(Brentford, United Kingdom), are also working on a
or his sore arm. Vaccines typically create antibodies
vaccine that induces an immune response against the
against the outermost portion of the hemagglutinin (HA)
stalk region of the HA protein. They’ve produced a
protein. That’s because our immune system recognizes
genetically-engineered virus that provokes a strong
the “head” of the protein most readily. Regrettably, that’s
antibody response against this stalk when tested in
also the spot that mutates most rapidly – meaning that
ferrets. The vaccine is now in Phase I clinical studies.
an immune response against it will typically only be good
for one flu season.
BEYOND THE STALK:
Scientists at BiondVax ( Jerusalem, Israel) are now INTERNAL PROTEINS
targeting the HA protein’s “stalk” instead. This bit of the
Not all efforts focus on the HA stalk. Other viral proteins
protein mutates much less frequently than the head.
also mutate somewhat infrequently. The trick is to
Slower mutation means the stalk is much more likely
identify which protein epitopes the immune system
than the head to remain the same from year to year.
recognizes. Scientists at Imutex (London, U.K.) think
BiondVax’s experimental vaccine, M-001, is a peptide
they’ve got it. The company’s FLU-v vaccine includes
vaccine. It consists of short stretches of the HA stalk
conserved, immune-inducing peptides from a range of
protein called “epitopes,” or sequences known to induce
viral proteins, including M1, M2, NP-A, and NP-B.
an immune response.

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The M2 protein is imbedded in the flu virus membrane Because mRNA molecules are relatively straightforward
and helps maintain the pathogen’s preferred internal to synthesize, this type of vaccine could incorporate
pH. The NP and M1 proteins surround viral RNA. Imutex mRNA encoding proteins from several different strains of
scientists determined which protein epitopes are most the flu, making it more universally protective. Moderna
likely to be displayed on the surface of the infected (Cambridge, MA) has an mRNA-based flu vaccine in
cell through a process known as antigen presentation. Phase 1 clinical development, while BioNTech (Mainz,
In this process, viral proteins inside of infected cells Germany) and CureVac (Tubingen, Germany) both have
are chopped up, and the resulting fragments are then mRNA flu vaccines in preclinical development.
“presented” or displayed on the cell surface, where they
elicit an immune response. Imutex is preparing TERM OF THE WEEK:
FLU-v for Phase III clinical studies.
PANDEMIC STRAINS
The word “pandemic,” is pretty scary, even if you didn’t
watch the dreadful 2016 movie of that name (we don’t
recommend it) or play the amazing board game or read
about the 1918 Spanish Flu Pandemic, which killed an
estimated 50 million people worldwide.
What is a pandemic? Simply put, it’s an outbreak of
disease that spreads rapidly over a wide geographic
area, affecting an exceptionally high proportion of the
population. Pandemic strains of the flu virus typically
result from a pathogen that normally infects animals
such as pigs or birds, but that mutate to infect people.
Since these strains are often brand new to the human
CAN MRNA SAVE THE DAY? population, few if any people have immunity. Thus, the
Another approach to generating a powerful immune illness spreads widely and rapidly. Protection against
response lies with viral messenger RNA (mRNA). mRNA is these potentially devastating strains is on the agenda of
the molecule that cells translate into a protein. When an any universal vaccine researcher.
mRNA encoding a viral protein is delivered to someone’s
Meanwhile, your best shot at protection from the
cells, they translate it into viral proteins. White blood
seasonal flu remains getting an annual flu vaccine from
cells then learn to recognize that protein, resulting in
your doctor, a health clinic, or even your neighborhood
immunity to the virus. Encouraging cells to produce
pharmacy. One day, and the sooner the better, we may
a viral protein very closely mimics natural infection.
get several years of protection against different flu
This pathogenic provocation should produce a strong
strains from just one needle. Now that sounds almost as
immune response – including by T-cells.
good as a pot of homemade chicken soup!

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CRISPR BABIES Life Science Training from Industry Experts

CRISPR BABIES Here at the WEEKLY, we’d decided to take the many
CRISPR headlines this week as a chance to review the
Earlier this week, a research team at the Southern
basics of how this technology works.
University of Science and Technology in Shenzhen,
China, announced the birth of twin girls whose genomes WHERE DID CRISPR COME FROM?
had been edited using CRISPR/Cas9. The work was done
CRISPR is a key immune response in bacteria. Like
at an in vitrofertilization clinic prior to implantation of
us, these microorganisms fall prey to viral infection.
days-old embryos. The targeted gene, CCR5, produces a
They’ve evolved a fascinating way to repel the invaders.
protein that the HIV virus uses to enter cells. Disrupting
In the 1980s, scientists observed a pattern in bacterial
this gene means that the virus will not be able to infect
genomes: repeating, palindromic sequences, with
the girls’ cells.
unique sequences — “spacers”— between repeats.
Not surprisingly, the work is controversial. Although They bestowed a tongue twister of a name, “clustered
CRISPR holds great promise for the treatment of genetic regularly interspaced short palindromic repeats,” on the
diseases, as with any experimental therapy, there mechanism, which we happily call CRISPR. Scientists also
are also risks involved. The U.S. National Academy of noticed CRISPR sequences always occur near genes that
Sciences has issued a report concluding that, while code for an enzyme that cuts DNA. This enzyme became
one day editing embryos at risk of serious genetic known as Cas, or “CRISPR-associated.”
diseases may be appropriate, at the present time the
In the mid-2000s, scientists realized these spacers
experimental technology should not be used in that
matched the DNA sequences of infecting viruses. The
manner. CRISPR has been used before to edit the
sick bacteria were stashing bits of the offending viral
genomes of human embryos by researchers in China as
DNA between its own CRISPR sequences! These viral
well as in the U.S., but this is the first time that an edited
DNA snippets create a “genetic memory,” which enables
embryo has been implanted and led to a live birth. In
the bacteria to fight back if reinfected. Here’s how:
the U.S., there is a law explicitly preventing the FDA
from approving medical treatments aimed at creating • Viral DNA present in the spacer sequences is copied
heritable genetic modifications. In China, the law is less into viral RNA.
explicit, but just days after the announcement of the • The bacteria make the DNA-cutting enzyme Cas,
CRISPR-modified babies, scientist He Jiankui is under which attaches to the new viral RNA.
investigation by the Shenzhen City Medical Ethics Expert
• The resulting viral RNA/Cas complex finds its match
Board, which claims to have no recollection of approving
on the invading viral DNA.
the work.
• The RNA attaches to the DNA and the Cas enzyme
CRISPR holds great potential for treating genetic
cuts up the foreign DNA, destroying the virus.
diseases, but most experts in genetic medicine and
bioethics are prescribing caution. The co-discoverers of • Voila — “healthy” bacteria.
the technology, Jennifer Duodna of U.C. Berkeley and
Feng Zhang of the Broad Institute, have both issued CRISPR EXPLAINED
statements urging a moratorium on the implantation of In 2013, researchers adapted this defense for use in
edited embryos until a global consensus on acceptable human cells. By adding a “guide RNA” and Cas enzyme to
risks is reached. This week, the second annual target a specific DNA sequence, scientists demonstrated
International Summit on Genome Editing is taking place the system could be used to cut human DNA in precise
in Hong Kong. Hopefully this meeting will be the starting locations! This original Cas protein came from the Cas9
point for reaching that consensus, so the scientific Streptococcus bacteria — hence the moniker CRISPR/
community can move forward and bring the life-saving Cas9.
potential of CRISPR to fruition.

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What makes CRISPR/Cas9’s ability to cut human DNA in By engineering double-stranded breaks at specific
precise locations so cool? The protein creates double- locations, scientists trigger the NHEJ or HDR cell
stranded breaks (DSB) in the specified DNA sequence. repair pathways.
Double-stranded breaks cut both strands of the • Activating NHEJ disrupts a disease-associated
DNA helix. gene. This prevents the production of the protein
Think of DNA as a two-lane bridge. Now imagine an that causes the disease.
earthquake takes place causing one section to break off, • Activating HDR fixes mutated genes by
falling away. DSBs can repair the damaged DNA bridge in simultaneously delivering a “repair template” that
two ways: contains the correct gene sequence.
• Homology Directed Repair (HDR) relies on a highly Both scenarios present possible cures for different
similar DNA segment to repair the break. In this case, types of disease. For example, CRISPR is currently in
workers build a new section of the genetic bridge Phase I clinical trials at Sichuan University (Sichuan,
offsite and then helicopter it into place. China). Here, researchers are delivering CRISPR/Cas9
• Non-Homologous End-Joining (NHEJ) closes components to cancer patients’ white blood cells to
the gap using another strategy. Visualize workers disable the PD-1 gene. The PD-1 gene inhibits these
pushing the two remaining sections of the bridge immune cells. By deactivating the PD-1 gene the immune
back together. NHEJ can result in a sequence error, system is left intact and the patient should be able to
just as sections of a repaired bridge often don’t line more aggressively fight cancer. Clinical researchers at
up properly. If the repair occurs in the middle of a the University of Pennsylvania (Philadelphia, PA) have
gene, it typically disrupts gene function and halts the also begun a Phase I trial using CRISPR to knockout PD1,
production of the matching protein. while CRISPR Therapeutics, in partnership with Vertex
Pharmaceuticals, has begun enrolling patients for a
Phase 1 trial using CRISPR to treat beta thalassemia. Next
on the research agenda come trials aimed at correcting
gene sequences mutated in hereditary blindness (Editas
Medicine, Cambridge, MA; expected 2019).

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BIOMANUFACTURING: HOW
BIOLOGICS ARE MADE Life Science Training from Industry Experts

PUTTING NATURE TO WORK Step two is for the manufacturer to establish a master
cell bank that supplies genetically identical cells for
There’s more than one way to skin a cat. Or make
future products. Companies create cell banks by
medicine, for that matter. It all depends on the kind—
transferring the production cell line to a bioreactor.
tabby or calico? Small molecule or biologic?
Though they may sound scary, bioreactors are simply
One key feature that distinguishes biologic drugs such vessels filled with a growth medium -- a “broth” with
as monoclonal antibodies from small molecule drugs the required nutrients brewing in optimal conditions
such as aspirin is their production. This highly complex of temperature, pH, and oxygen concentration for
process, biomanufacturing, requires much more time and cell growth.
expense than needed for small molecules.
The cells are left to simmer, or multiply for a few
In this two-part WEEKLY series, we’ll revisit key generations, creating hundreds of millions of identical
components of biomanufacturing. First, we look at cell copies. The manufacturer collects this slough and
bank production and examine the different types of cells portions them into small vials. Each of the several
used in biomanufacturing. Next week, we zoom in on the hundred receptacles contains about a million (million!)
step-by-steps of biomanufacturing. cells. The vials are then frozen with liquid nitrogen,
cooling them to -196 degrees Celsius. The deep freeze
TERM OF THE WEEK: stops cell growth; In other words, if some future scientist
BIOMANUFACTURING thawed one of the vials in twenty years, she or he would
Biomanufacturing is the production of biological find the cells inside exactly as they were at storage—
products from living cells. Companies use the process barring apocalypse or someone tripping over the power
to make biologic drugs such as antibodies and enzyme strip. This stable longevity is key, as product consistency
replacement therapies. Small molecule drugs can over the lifetime of the product is critical to drug safety.
be synthesized chemically. As their name suggests, Manufacturers typically divide the master cell bank for
biologics require living cells. storage in three separate locations so that disaster in
Biomanufacturing isn’t just about medicine though. For one place doesn’t wipe out this important resource.
instance, companies use the process to make enzymes In each location that a product is manufactured, a
for bioremediation—that is, cleaning up toxic stuff in the manufacturer creates a working cell bank by thawing
environment. The food processing industry also uses one vial from the master cell bank and “expanding it,” or
biomanufactured products. allowing it to multiply for a few generations -- and then
freezing several hundred vials for storage. Each new
Step Number One: Establishing the Cell Bank
biomanufacturing campaign starts by thawing a vial of
First things first. Biomanufacturing involves engineering cells from the working cell bank.
a cell to produce a specific protein. Using well-
established techniques, scientists transfer a gene
encoding the desired protein into a “production cell.”
The two most commonly used production cells are E. coli
bacterial cells and Chinese hamster ovary cells, or CHO
cells. Once a manufacturer successfully manipulates a
cell to produce said protein, the cells multiply. Scientists
call these genetically identical cells the production
cell line.

Step Two: The Master Cell Bank

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MAMMALIAN VS. BACTERIAL HAMSTER OVARIES? WHAT???
The first biologic drug, insulin, was produced using E. Why Chinese Hamster Ovary cells? Well, since you had
coli cells. Researchers soon realized, however, that they to ask… When scientists first realized they needed some
couldn’t produce every therapeutic in bacterial cells. cell type other than E. coli to produce complex biologic
Highly complex proteins, such as monoclonal antibodies drugs, CHO cells were handy. Scientists worldwide
and certain enzymes, present two main obstacles. were already using them in many experiments and they
Bacterial cells are unable to correctly fold these complex made a convenient platform for biologics. More than
proteins, nor are they able to confer required post- thirty years of data has made it clear it’s safe to produce
translational modifications - chemical and physical drugs in these cells. As a result, the FDA has granted
changes made to a protein by cellular enzymes after the them “generally-regarded-as-safe” (GRAS) status for
protein is produced. therapeutic protein production. That is, drug companies
can use them to manufacture product without first
NO, NOT THAT E. COLI demonstrating their safety.
Life-saving drugs grown in E. coli bacteria seems a bit Of course, it’s important to remember that no matter
sketchy, does it not? The same bug that causes food what type of cells are used to produce the therapeutic
poisoning? Not exactly. While it sounds unsavory, it’s protein, the “nursery” cells don’t make it into the final
important to remember that there are many different product. After manufacturers grow therapeutic protein-
strains of E. coli, most of which are benign. In fact, E. coli producing cells for several days or weeks, the next step
bacteria make up a big chunk of healthy gut microbiota. is to purify the therapeutic protein away from other
cellular proteins and the cells themselves. Look for more
in Part 2 of this series next week!

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WADING DEEPER INTO BIOMANUFACTURING Life Science Training from Industry Experts

WADING DEEPER or other microorganisms. To coin a phrase, one bad

INTO BIOMANUFACTURING microbe spoils the bunch. The FDA has established
very strict protocols for maintaining aseptic
Last week, we began our exploration of
manufacturing conditions.
biomanufacturing--defining terms, introducing
the different types of cells used in production, and DOWN THE STREAM: HARVEST,
explaining how manufacturers create cell banks to PURIFICATION, FORMULATION
provide raw materials for the medicines some of us know
The downstream side of biomanufacturing takes the
and love. This week, we turn to how those cells ultimately
scaled-up “stuff” from the bioreactors and turns it into
result in finished products.
the end product.
UP THE STREAM: SCALE UP & Harvest
TESTING, TESTING, TESTING
Once production cells reach maximum density, it’s
Like biotechnology, manufacturing has its own lingo. harvest time. Different types of drugs require different
For instance, the overall manufacturing process can be harvesting techniques. Most biologics, such as
described in terms of “upstream” and “downstream” antibodies, insulin, and growth hormones, are secreted
activities. Think of upstream like this: getting the stuff from the production cells right into the growth media.
necessary to make stuff to sell. Thus, the first step in harvesting our therapeutic
Once technicians thaw a vial of cells from a working proteins is to separate the growth medium from its vat
cell bank, scaling up can begin. Scaling up, in this mates—aka the production cells. This is done by high-
case, means transferring cells into successively larger speed centrifugation, which physically separates the
bioreactors to accommodate enough growing cells for solid cells from the liquid growth media.
production. No scale up, no product. That’s because cells
require a certain critical density to thrive. Just pouring a
Separation Anxiety
smallish vial of cells from a master cell bank into a 20,000 The growth medium contains a “soup” of proteins – not
liter vat won’t cut it. only the therapeutic protein that the production cells
were engineered to produce, but also other proteins
A final bioreactor volume varies in size--anywhere from
secreted by the cell, as well as proteins added to the
3,000 liters to 100,000 liters. It all depends on the type of
growth medium as nutrients. The manufacturer needs to
production cell and the desired amount of end product.
separate the therapeutic protein from all of these other
For example, a typical manufacturing campaign based
proteins. Column chromatography is used to accomplish
on CHO cells might last two weeks and result in a final
this purification. This process teases out individual
volume of 20,000 liters. In contrast, a standard E. coli
proteins based on size, structure, or electrical charge.
campaign takes only a few days, yielding an ultimate
They all pass through a cylindrical column packed
volume of 80,000 to 100,000 liters. The differences in
with a solid resin that catches different proteins. The
time and final volume stem from how quickly different
proteins travel the column under gravity and with the
cell types divide. Under ideal conditions, E.coli splits once
application of moderate or high pressure (fast protein
every 20 to 30 minutes or so, while the pokier CHO cells
liquid chromatography or FPLC) or high pressure (high
divide about once a day.
performance liquid chromatography or HPLC) to increase
Along the way, manufacturers examine cell viability, speed and resolution. Some of them flow through the
product concentration, and product activity. They column; others remain inside—to be washed off (eluted)
also monitor the physical environment to optimize under different conditions.
temperature, pH, nutrients, and oxygen. Finally, it’s
For example, isolating a negatively-charged protein
critical to test for contamination by bacteria, yeast,
requires a resin with positive charges. When a protein

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soup goes into a “positively-resined” column, the FORMULATION, FILL, AND FINISH
neutral and positively-charged proteins pass through;
After purification, the manufacturer formulates the
the resin snares those that are negatively charged and
therapeutic protein according to its established research
makes them stay put. These “captured” proteins are
and development specifications. These requirements
later washed off the column with a salt solution--the
take into account factors such as product stability
positively-charged sodium ions peel the negatively-
and delivery method. The company may use product
charged proteins from the resin. The proteins pulled
excipients--pharmacologically inactive ingredients to
off the column in this wash step are collected for
enhance the drug product. These may include color
further processing.
additives, time release factors, and bulking agents such
as mannitol. The manufacturer may also add ingredients
that increase a product’s stability, such as antioxidants,
buffers, and surfactants (to decrease clumping). Finally,
the company establishes product fill concentration, adds
the appropriate label, and packages the product for
large-scale distribution.
From engineering a production cell line to creating cell
banks, then growing up large volumes of therapeutic
protein-producing cells, to finally purifying and
formulating the final product, biomanufacturing is a
highly complex process. And of course, all of these
Of course, the protein soup contains “extra” proteins
steps must be carried out with careful attention to
other than the target protein that bear a negative charge.
following current Good Manufacturing Practices (cGMP).
Typically, the proteins captured by the positively-charged
A thorough understanding of the process and variables
resin will next be loaded onto another type of resin that
that may affect it are critical to any company embarking
sifts them out based on other characteristics such as
on biologics production.
shape or size. In some cases, a third round of this protein
straining is necessary. Each therapeutic requires its own
specific purification protocol.

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STUMP THE CLUMPS: NEW DRUGS
TO TREAT AMYLOIDOSIS Life Science Training from Industry Experts

STUMP THE CLUMPS: NEW DRUGS the nerves that assist in organ function, digestion and
TO TREAT AMYLOIDOSIS kidney complications follow.

We at the WEEKLY aim to keep you up on the coolest, Let’s look at the new drugs approved to prevent
latest innovations in biotech. Bonus: we might also help this problem.
you win Jeopardy. Ready? The category is Rare Diseases
for $1,000. The clue is “Amyloidosis.”
ONPATTRO: GENE SILENCING IS GOLDEN
The first drug approved to treat polyneuropathy
Ding ding ding! Alex, what is a group of diseases caused
associated with hATTR was Alnylam Pharmaceutical’s
by the abnormal buildup of proteins in tissues and
(Cambridge, MA) Onpattro in August 2018. Onpattro
organs? You win!
was also the first FDA-approved drug to work by RNA
Maybe some reader out there will win Jeopardy. interference (RNAi), or gene silencing. This approach
Meanwhile though, it’s the patients who suffer from aims to reduce how much protein a specific gene
maladies associated with these protein clogs and their produces – in this case, the mutated transthyretin gene.
families who are really poised to win. Because last year, Onpattro does this by destroying the gene’s RNA – recall
the FDA approved two new drugs to treat hereditary that the information in genes is first converted to RNA,
transthyretin amyloidosis (hATTR). 2019 may see another which is then translated to a protein. Less mutated
approved as well. This week, we explore the science transthyretin protein means fewer amyloid fibrils, which
behind this uncommon but debilitating illness, and look means reduced polyneuropathy.
at the brand-spanking new treatments for it.
How does gene-silencing work? RNAi takes advantage
First, more about amyloidosis. What the heck are of existing cellular pathways that target and destroy
amyloids? Short answer: proteins that build up in organs double-stranded RNA (dsRNA).
like the liver, spleen and in other tissues. Scientists
To activate the pathway, researchers introduce a
call the collections amyloid fibrils--clumps of proteins
double-stranded or “hairpin” shaped RNA. In the case
that fold into a shape that allows many copies to
of Onpattro, this RNA hairpin is the drug. The enzyme
stick together.
DICER cuts it up to produce a “short interfering RNA”
Many different proteins form amyloids. Perhaps the best (siRNA). The siRNA binds to a second enzyme, RNA-
known comes from amyloid-beta, and is associated with induced silencing complex (RISC). RISC alters the RNAi
Alzheimer’s disease. so that only a guide strand which is complementary to
the target RNA remains. This guide strand/RISC complex
A NERVE-WRACKING PROBLEM then attaches to the problematic transthyretin mRNA
The protein transthyretin can also cause amyloidosis. and destroys it.
Produced in the liver, transthyretin transports thyroid
hormones and vitamin A throughout the body.
Mutations cause transthyretin to form amyloid fibrils,
which accumulate, especially in the heart and nervous
system. These deposits result in a range of symptoms,
including cardiomyopathy--congestive heart failure
and abnormal heart rhythms. Amyloid accumulation
in nerves can cause tingling, numbness, or burning
pain anywhere in the body. It also sometimes leads to
weakness and loss of mobility. Doctors call this assembly
of problems polyneuropathy. If protein clumps damage

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Patients receive Onpattro through IV infusion every Patients self-administer Tegsedi weekly. This is more
three weeks. convenient than the Onpattro infusions. However, some
patients and doctors may prefer the Alnylam medication
TEGSEDI: STOP MAKING SENSE due to its better safety profile.
Tegsedi, from Ionis Pharmaceuticals (Carlsbad, CA)
and Akcea Therapeutics (Boston, MA), was the second NEXT IN LINE: TAFAMIDIS
drug approved for hATTR-associated polyneuropathy, The European Medicines Agency (EMA) has approved
in October 2018. Tegsedi is an antisense drug. Like Pfizer’s (New York, NY) drug tafamidis for hATTR
Onpattro, antisense drugs activate a cellular pathway polyneuropathy. The FDA is expected to okay the
that destroys a targeted RNA. They are short, synthetic drug in 2019 for hATTR cardiomyopathy. Unlike the
pieces of RNA whose sequence complements the RNA previous treatments which act on RNA, tafamidis is a
that codes for a disease-associated protein. When the pharmaceutical chaperone. In other words, it’s a small
drug enters a patient’s cells, it binds to the disease- molecule drug that helps transthyretin fold correctly,
causing RNA. This binding triggers an enzyme called resulting in fewer garbled proteins to form amyloid
RNAse H to wipe out the antisense-target RNA duo. Once fibrils and cause disease. Like most small molecule
again, no RNA means no protein to form amyloid fibrils. drugs, it is administered orally.

NON-HEREDITARY
AATR CARDIOMYOPATHY
Although hAATR is very rare – estimated to affect about
50,000 people worldwide – non-mutated transthyretin
forms amyloid fibrils in ten to twenty percent of the
elderly, contributing to cardiomyopathy.
Next week, we’ll widen our focus from a rare disease
to a growing public health epidemic: Nonalcoholic
Steatohepatitis (NASH).

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THE RACE AGAINST NASH Life Science Training from Industry Experts

FIGHTING A SILENT EPIDEMIC WHEN WHITE CELLS RUN AMOK

Consider the liver. It’s just one of the jumble of stuff Livers can only handle so much fat. Too much, and liver
inside that makes us tick, right? What do you really know cells release inflammation-inducing signaling molecules,
about it though—other than it’s “vital”? In fact, the liver known as cytokines. These proteins trigger a series
is your largest internal organ, and plays vital roles in of events, the end result of which is the Attack of the
neutralizing toxins, fighting infections, manufacturing White Blood Cells. We think of these hemocytes as
proteins and hormones, controlling blood sugar, and “good,” but not so with NASH. White blood cells, such as
helping to clot the blood. It may just be time to start macrophages, invade the liver, increasing the chances
giving our livers a little more thought. We don’t just that liver cells will die and damaging scar tissue will form.
mean by taking it easy on the cocktails either. Liver • Boehringer Ingelheim’s (Ingelheim, Germany)
disease is a growing health concern worldwide. A big investigational NASH drug, BI146735, is a small
part of the problem is that that the most common liver molecule inhibitor of the vascular adhesion protein 1
disease in developed countries, non-alcoholic fatty (VAP1). VAP1 helps white blood cells to migrate into
liver disease (NAFLD), often has no symptoms. Its onset the liver. Inhibiting VAP1 reduces this migration. Ta-
is associated with obesity and type 2 diabetes—two da—decreased inflammation! BI146735 in in Phase
conditions that are also on the rise. II clinical studies. BI146735 was originally discovered
In 2017, NAFLD was estimated to affect nearly one by Sydney, Australia-based Pharmaxis.
quarter of people worldwide. About 30 to 40 percent of • Gilead’s (Foster City, CA) small molecule drug
people in the United States are thought to be affected, selonsertib, in Phase III trials, takes a slightly
and about 3 to 12 percent of American adults suffer different approach to inflammation. It inhibits
from the advanced form of the condition, non-alcoholic the activation of two enzymes involved in cellular
steatohepatitis (NASH). Currently, its only treatment is pathways leading to inflammation, liver cell injury,
liver transplantation. However, a number of biopharma and scarring. Gilead is also testing selonsertib in
companies are tackling this growing concern. Let’s take a combination with its Phase II NASH candidates,
closer look. GS-0976 and GS-9674. The first is a small molecule
that puts the brakes on an enzyme involved in
EASILY CONFUSED: NAFLD VS. NASH producing fatty acids. The other is an FXR-nuclear
NAFLD occurs when excess fat accumulates in the liver receptor inhibitor.
of people who drink little to no alcohol. NASH, the more
• Allergan’s (Dublin, Iralend) cenicriviroc takes aim
severe form of the disease, is characterized by liver
at liver inflammation by inhibiting receptors on
inflammation and scarring. As scar tissue accumulates,
the surface of white blood cells called chemokine
it impairs liver function. It’s the number one cause of
receptors. Chemokines are chemical messengers
non-alcohol related cirrhosis (severe, late stage scarring)
that stimulate movement of cells towards the
which in turn can lead to liver failure.
source of their release—typically damaged or
People with diabetes, obesity, or metabolic syndrome infected tissue. This chemokine APB often helps
run the risk of developing NAFLD and eventually, the body fight infection. However, with NASH, this
NASH. Treatments in the works largely focus on just damages the already inflamed liver further.
reducing inflammation and improving how the liver Cenicriviroc is in Phase III clinical studies.
metabolizes fats.

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FIGHT THE FAT Researchers hope this change may disrupt the
progression of NASH.
An alternate approach to treating NASH works on
improving a patient’s lipid metabolism. Control the fat • Yet another nuclear receptor activator is Genfit’s
that accumulates in the liver, control the disease. (Loos, France) investigational elafibranor. This drug
works on the receptors PPAR α/σ. Activating these
• Intercept Pharmaceuticals (San Diego, CA) has
receptors switches on genes that increase the
begun Phase III clinical studies on Ocaliva—a small
metabolism of fatty acids, decreasing liver fat and
molecule drug. This product works by binding the
improving lipid profiles—as well as increasing insulin
nuclear receptor FXR. This is a type of receptor
sensitivity and anti-inflammatory activities. The
protein that is present inside of cells, rather than
drug in now in Phase II clinical studies for advanced
on their surface, like most other receptor proteins.
NASH patients.
When activated by the appropriate signaling
molecule, the nuclear receptor moves inside the Several other NASH drugs are coming down the pike,
cell’s nucleus, where it binds DNA at a specific including Galmed’s (Tel Aviv, Israel) Aramchol (inhibits
location, turning on the expression of particular fatty acid synthesis; Phase II) and San Diego-based
genes. Ocaliva specifically binds to and activates Conatus’ Emricasan (inhibits inflammation; Phase II).
FXR, which modulates the expression of genes Let’s hope that 2019 sees at least one FDA approval from
involved in lipid metabolism and glucose regulation. the handful of Phase III candidates out there to step up
the fight against this silent epidemic.

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AN EGG-CELLENT IDEA Life Science Training from Industry Experts

AN EGG-CELLENT IDEA can researchers ensure that the drug is only produced
in egg cells? By including an egg-specific promoter with
“The incredible, edible egg.” This advertising slogan
the gene that is transferred to the eggs. A promoter is
created for the American Egg Board in the 1970s rings
a stretch of DNA that controls whether or not a gene is
truer than ever. Here’s why: scientists at the Roslin
turned on or off in a specific cell type. An egg-specific
Institute (Midlothian, Scotland), part of the University
promoter turns on the gene only in egg cells.
of Edinburgh, recently published a paper describing
a novel, potentially cost-saving approach to the There’s also some good news for animal lovers. Life
production of biologic drugs. The crucial element? Eggs. proceeds as normal for the birds– maybe even a little
One estimate states that the new process could make better. These very valuable, genetically enhanced
manufacturing biologics one hundred times cheaper chickens receive extra special care. They just go about
than existing methods. But how? their egg-laying business, completely unaware that their
ova may in fact be golden.
Take some chickens, fiddle with their genetics so that
they lay eggs containing human proteins that fight MORE THAN JUST CHICKENS
cancer. This WEEKLY provides some details.
The drugs described above are not the first medicines
BREEDING FEATHERY BIOREACTORS brought about through animal husbandry. In 2009,
the FDA approved ATRyn, a recombinant version
Roslin Institute scientists genetically modified chickens
of the antithrombin protein. This anti-clotting and
to lay eggs with either the protein interferon alpha-2a
anti-coagulation factor is produced in goat milk.
(IFN2a) or macrophage-CSF (M-CSF). Both are cytokines,
Manufactured by rEVO Therapeutics (Framingham, MA)
a type of immune system signaling molecule. IFN2a
ATRyn treats certain blood clotting disorders.
treats hairy cell leukemia, malignant melanoma, AIDS-
related Kaposi’s sarcoma, and other cancers. The In 2015, the FDA approved Kanuma (Alexion
interferon packs a two-fisted anti-cancer punch. First, Pharmaceuticals; Boston, MA), a recombinant version
it interferes with malignant cells’ ability to divide (this is of lysosomal acid lipase. This enzyme helps break down
actually the origin of the name “interferon”). Second, it fatty acids. The drug is produced in rabbit milk. It was
revs up the immune system to fight cancer. The other approved to treat lysosomal acid lipase deficiency, an
cytokine, M-CSF, is being investigated as a therapy to inherited disease which affects a number of organs.
stimulate damaged cells to repair themselves. ATRyn and Kanuma are medicines made in a small
To get the hens to lay biologic-producing eggs, Roslin quantities for rare diseases. The Roslin group’s platform
scientists deliver genes for each protein to chicken may be better suited to producing large quantities of
embryos via a by viral vector. Viral vectors are viruses drugs, due to the prodigious number of eggs one hen
that have been designed to deliver a specific gene produces annually—over three hundred! Chickens also
to a cell, incorporating it into the target cell’s DNA. require less room and board compared to goats or
Integrating the gene for the protein into an embryo rabbits. Lastly, there’s a lot of protein in each egg white.
means it ends up in all of the embryonic cells, including Three eggs contain enough cytokines for one dose
those for the egg. of medicine.
No matter what animal, remember that neither its egg
KEEPING THE PROTEINS IN CHECK nor milk is the actual drug. Just as in “old-fashioned”
Scientists want the chicken to produce the new protein biomanufacturing, therapeutic proteins need to be
only in her egg cells. Cytokines can be powerful purified through column chromatography before they
medicine. That means that they can actually harm an can be used in medicine.
animal by creating an overactive immune system. How

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FLU VACCINE—A DIFFERENT COCKTAIL FODDER: REMEMBER DOLLY?
KIND OF EGG-CELLENCE Roslin Institute scientists have a history of making
You may have heard about flu vaccine produced in eggs. scientific breakthroughs. In 1996, the famous cloned
Cooking up flu vaccine, however, differs from making sheep, Dolly, was created there. Dolly was the first
IFN2a or M-CSF. Instead of tweaking a chicken embryo’s mammal to be cloned from an adult cell.
genetics, technicians inject different strains of flu virus
into eggs to grow. Once enough virus has propagated
inside the egg “cocoon,” it’s removed, purified,
inactivated, and formulated into vaccines.

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AN EPIGENETIC LIQUID BIOPSY Life Science Training from Industry Experts

A NEW TYPE OF LIQUID BIOPSY The end result: methylation reduces or even blocks
gene expression. Areas of the genome that have
Biopsy. It can bring to mind painful cutting, punching and
methyl groups attached to them are referred to as
poking. Thankfully, these important and potentially life-
methylated. DNA methylation is a normal part of cellular
saving procedures have moved into the new millennium
development. However, variations in the “normal”
along with the rest of us.
methylation pattern are associated with disease.
Liquid biopsies have become a hot area of development
in cancer diagnostics. These tests arise from the simple THE CANCER METHYLSCAPE
fact that tumors shed DNA and cells. It’s possible to Last fall, researchers at the University of Queensland
detect these tumor leftovers in bodily fluids such (Brisbane, Australia) published work describing a
as blood, urine, or saliva. So liquid biopsies present “methylscape” (methylation pattern) that is common
a much more attractive alternative than traditional to most cancers. They found that in many cancer cell
and often repeated tissue biopsies to screen for genomes, regulatory regions—which help control gene
cancer and monitor treatments. However, one major expression—are methylated significantly more than
drawback exists. Current versions use pricey, lab- normal. In contrast, other parts of the genome display
based technologies such as DNA sequencing to detect much less methylation than normal.
cancer markers.
Such differences in methylation patterns change the
Fortunately, there may soon be a new liquid biopsy in structure of DNA. This, in turn, determines what else
town. The new tests detect epigenetic markers, rather interacts with the DNA. In their study, the University of
than genetic mutations. They could prove to be less Queensland team found that DNA that exhibits a cancer
expensive, faster, and easier to administer. methylscape binds and sticks to gold nanoparticles much
better than DNA from healthy cells.
TERM OF WEEK: EPIGENETICS
Epigenetic modifications are changes to DNA that A GOLDEN TEST
don’t alter the actual gene sequence themselves, like Any identifiable difference between healthy and
mutations. Instead, they are chemical modifications unhealthy tissue can provide a basis for a diagnostic
to the DNA. These alterations typically affect gene test. Find the disease-associated variance, diagnose the
expression, that is, how often a gene gets read by illness. The University of Queensland researchers have
the cell. used the gold nanoparticle affinity difference as the
One of the most common types of epigenetic basis of a potential new way to identify the presence
modifications is methylation—the addition of a methyl of cancer from cancer cell DNA that may be present
(CH3) group to cytosine (C) nucleotides. in a blood or other fluid sample—i.e., a liquid biopsy.
It’s officially called the “salt-induced gold nanoparticle
aggregation system,” or gold nanoparticle test for short.
It relies on the fact that adding salt to a gold nanoparticle
solution causes the nanoparticles to aggregate, resulting
in a color change from reddish to blue. If DNA is
attached to the nanoparticles, however, no aggregation
occurs, and thus no color change. This test is used to
differentiate between cancer cell DNA and healthy DNA
as follows:
• DNA is added to the nanoparticle solution.

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 • Cancer methylscape DNA sticks to gold huge advantage over other biopsies. It could make
nanoparticles, preventing aggregation; DNA from an ideal screening for many sorts of cancer and for
healthy cells doesn’t. monitoring treatment.
• In the next step, salt is added. If the nanoparticles Other advantages include the test’s 10-minute
fail to aggregate (cancer DNA), the solution’s color turnaround time and lower cost. Despite its reliance on
remains the same. If they do (healthy DNA), the gold, researchers anticipate a significant cost savings.
solution’s color changes. That’s because the test uses a miniscule amount of the
Using this technique, the University of Queensland team shiny stuff, especially in comparison with the cost of
identified malignant cell-derived DNA with a stunning current liquid biopsies that rely on DNA sequencing.
90 percent accuracy. They examined more than two Finally, the new test is much more portable than large
hundred tissue and blood samples. The types of cancers sequencing machines, meaning that technicians may be
tested included breast, prostate, bowel and lymphoma. able to administer it right in a doctor’s office.
The methylation pattern that underlies the test is The researchers are working with UniQuest, the
consistent across most types of cancer, so it’s likely that University of Queensland’s commercial partner,
the test could detect many other cancers as well. to identify a commercial partner and move into
clinical testing. Only time will tell, but this innovative
UNIVERSAL, INEXPENSIVE, new test could dramatically change the cancer
FAST, PORTABLE! diagnostics landscape.

There is potentially much to love about the new
gold nanoparticle test. Its universality gives it a

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PARP1 INHIBITOR TAKES ON
PANCREATIC CANCER Life Science Training from Industry Experts

PARP1 INHIBITORS would normally do their job are prone to sustaining

DNA damage at a much higher rate than normal. This
PARP1 inhibitors are back in the headlines this week,
higher rate of DNA damage increases the chances of
as Astra Zeneca’s (Cambridge, England) and Merck’s
cancer developing in those cells. BRCA1/BRCA2 positive
(Kenilworth, New Jersey) Lynparza posted positive Phase
cancer is cancer that is associated with mutations in the
III results from a trial with pancreatic cancer patients—a
BRCA1/BRCA2 genes. The mutations are most strongly
notoriously difficult cancer to treat. Lynparza has been
associated with breast and ovarian cancer, but are also
shown to help patients with BRCA-mutated metastatic
associated with increased risk of developing stomach,
pancreatic cancer live longer without their cancer
pancreatic, prostate, melanoma, leukemia, lymphoma,
progressing. The drug has already gained two FDA
and colon cancer.
approvals for BRCA-mutated breast and ovarian cancers.
This week, we’ll take a look at what PARP1 inhibitors are THE POINT OF PARP
and their connection to BRCA-mutated cancers.
PARP1 is a DNA repair protein. By stopping the PARP1
DNA DAMAGE RUNS DEEP repair pathway in cells already deficient in BRCA1/
BRCA2-mediated repair, cancer cells become extremely
PARP1 inhibitors work by exploiting the cellular pathways
vulnerable to DNA damage. Because of this, DNA damage
found in DNA damage repair. So, how exactly does DNA
accumulates and triggers apoptosis. A PARP1 inhibitor is
get damaged?
usually administered in combination with chemotherapy
DNA incurs approximately 10,000 to 1,000,000 or radiation therapy, which increases the incidence of
“molecular lesions” per day from breaks or “nicks” to the apoptosis-triggering DNA damage. Healthy cells, which
double helix, or chemical modification to the A, C, G, or still have BRCA repair pathways intact, are less sensitive
T bases. This may sound high—but remember, our DNA to additional DNA damage.
contains six billion bases (three billion base pairs), so this
is equivalent to .001% to .1% of the total DNA in each cell.
This damage occurs as a result of normal DNA replication
errors and environmental exposures, such as ultraviolet
radiation, X-rays, and chemicals.
The good news is our cells have mechanisms to fight
against this damage before it causes harm. DNA repair
proteins find and fix different types of DNA damage.
If DNA damage exceeds a threshold amount (beyond
which repair is possible) a protein called p53 triggers cell
death—also known as apoptosis. DNA repair proteins
prevent errant cells from turning into cancerous cells,
a likely outcome if the damage accumulates in genes BEYOND CANCER
important for regulating cell growth and division. Preclinical research suggests that PARP1 inhibitors
may also be relevant to other disease areas, such as
BEHIND BRCA autoimmune and inflammatory disorders. PARP1 has
Arguably the most famous DNA repair proteins, BRCA1 been shown to play a role in activating proteins that
and BRCA2, were first discovered to be active in drive inflammation. Preclinical models demonstrate
breast tissue, hence the moniker “breast cancer type that in cases without the PARP1 gene, subjects were
1/2 susceptibility,” or BRCA. If these repair proteins less vulnerable to rheumatoid arthritis than with the
themselves are non-functional, the cells in which they gene. Inhibiting PARP1 resulted in reduced signs of

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inflammation in models of multiple sclerosis, irritable always pairs with a “G” base. These base-pairing rules
bowel disease, and allergic airway inflammation. are what enable DNA to replicate faithfully from one
generation of cells to the next. However, uncorrected
EASILY CONFUSED: DNA DNA damage may cause that “A” base to mistakenly pair
DAMAGE VS. DNA MUTATION with a “G” during replication; or a “C” to pair with a “T.”
BRCA1, BRCA2, PARP1, and other DNA-repair proteins This results in a sequence change —a mutation—in the
correct DNA damage, but they don’t fix mutations. replicated DNA. The gene now provides incorrect genetic
What’s the difference? information to the cell.

DNA damage refers to alterations in the chemical

structure of DNA. This may mean a break in the
DNA strand, a substitution to one of the bases that
make up DNA (A, C, G, or T), or even a missing base.
These changes are detected and corrected by DNA
repair enzymes.
A DNA mutation is a change to the actual base sequence
(A, C, G, or T). Mutations can arise if DNA damage
is not corrected. Recall that in undamaged DNA, an
“A” base always pairs with a “T” base, and a “C” base

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THE MOLECULAR CAUSES OF OBESITY Life Science Training from Industry Experts

A BIG FAT PROBLEM We all know obesity is bad for us—it’s associated with
a slew of health problems, including non-alcoholic fatty
Bacon-wrapped meatloaf, chicken and waffles, cronuts! liver disease, hypertension, coronary heart disease, and
Mmm…but no wonder some of us are getting a little stroke. It’s also the primary cause of type 2 diabetes.
pudgy. According to the Center for Disease Control
(Atlanta, GA), 40 percent of adults and nearly 20 percent
THE FACTS OF FAT’S LIFE
of children and adolescents in the U.S. are obese.
Fortunately, the pharmaceutical industry is working Traditionally, fat was thought of as a way to store energy.
on it. The body converts extra calories into adipose tissue, aka
fat. The fat can then be broken down into fatty acids,
Current drug interventions attempt to limit the amount
serving as fuel when needed. Now doctors, nutritionists
of food patients eat, which has led to good results for
and other believe that adipose tissue plays a more
some. However, a large need still exists for therapeutics
complicated role in our bodies’ routines.
that target the underlying molecular causes of obesity—
starting with adipose (fat) tissue.
THE DREADED MUFFIN TOP
In this WEEKLY, we explore the emerging view that GETS MORE DREADFUL
adipose tissue is a metabolic organ that undergoes
Researchers have made a critical discovery. They’ve
pathological changes when we become obese. We also
found that where we gain weight matters too: increased
introduce some of the new products researchers are
adipose tissue within our abdomen is linked to metabolic
developing to help fight the fat.
disease. Not so with the fat just beneath our skin.

TERM OF THE WEEK: OBESITY So what happens as abdominal adipocytes get bigger?
Short answer: fat cell communication runs amok.
The word “obese” isn’t just another way of saying
Whaaa?
somebody is fat. In fact, it’s a medical term. The public
health community defines obesity as a medical condition Yes, people, our fat cells talk to our bodies. Adipose
in which a person has accumulated so much body tissue has its chemical messengers, called adipokines.
fat that it will likely affect his or her health. Clinically, These metabolic signaling molecules allow adipose tissue
doctors consider someone obese when their body mass to communicate with our brain, liver, immune system,
index (BMI) is over 30 kg/m2. (If you’re curious, you can and other organs.
calculate your own BMI by dividing your weight by the
square of your height.) AN INFLAMMATORY CASCADE
Interesting factoid: the number of the cells that make Getting fatter changes our adipokines. These molecules
up adipose tissue—adipocytes—remains the same once normally suppress inflammation. When we gain fat,
we reach adulthood. When we gain weight, we don’t adipokines turn on us to become molecules that
gain more adipocytes. Instead, they just get bigger. This promote inflammation. The inflammation then attracts
adipose-cell elasticity may partially explain why it’s hard macrophages, a kind of white blood cell, to the fat tissue.
to keep those pounds off. Fat cells are always there, just They promote even more inflammation, which in turn
waiting for a refill. results in decreased insulin sensitivity throughout the
body. Decreased insulin sensitivity means less ability to
Obesity is more than a case of too many cheeseburgers.
regulate blood glucose levels, and is a hallmark of type
It typically stems from a combination of lifestyle and
2 diabetes.
genetics, although a few forms do result entirely from
genetics. Some people can lose and control their weight When adipocytes “fatten up” they outpace the ability of
by changing their diet and increasing how much they the surrounding blood vessels to feed them. This results
exercise. However, maintaining a healthy size is so in low oxygen levels in the fat tissue. This “asphyxiation”
difficult that most people regain any lost weight within a leads to even more inflammatory adipokines. This
few years. promotes yet more insulin insensitivity.

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What does that mean for our health? Ultimately, • TNF inhibitors: One of the key drivers of
abdominal adipocytes become so dysfunctional that inflammation is the signaling molecule TNF.
they can grow no bigger. Then fat begins to accumulate Remicade ( Janssen Biotech, Horsham, PA) and
where it doesn’t belong. It shows up in the liver, skeletal Enbrel (Pfizer, New York City, NY) both inhibit TNF.
muscle, and pancreas and disrupts their functions. Though already approved for other inflammatory
In the pancreas, it interferes with insulin production, conditions, preliminary studies suggest that
potentially leading to diabetes. In the liver, it’s associated both drugs may help with glycemic control and
with non-alcoholic fatty liver disease. Finally, in skeletal insulin resistance.
muscles, misplaced fat can impede mobility. • IL-1 beta inhibitors: Another important pro-
This deeper understanding of how adipose tissue inflammatory signaling molecule is IL-1 beta.
functions has led to some new pharmaceutical Anakinra is an IL-1 beta inhibitor developed
approaches to treating obesity. The therapeutic goal: by Swedish Orphan Biovitrum (Stockholm,
to reduce some of the systemic effects such as insulin Sweden). It’s being studied to treat obesity and
resistance and inflammation. type 2 diabetes. Anakinra is already approved for
rheumatoid arthritis. Novartis’ (Basel, Switzerland)
ADJUSTING ADIPOKINES IL-1 beta inhibitor Ilaris is also being investigated for
treating insulin resistance.
Adiponectin is one of the adipokines which normally
increases insulin sensitivity and decreases inflammation. • Blocking macrophages: Macrophages move into
Researchers at the Catholic University of Korea adipose tissue in response to signaling molecules
(Seoul, Korea) are conducting preclinical testing of called “chemokines.” Researchers think they may be
AdipoRon, a small molecule activator of the adiponectin able to block macrophages by inhibiting chemokines,
receptor present on the surface of muscle and liver and so disrupt the cycle of inflammation.
cells. Scientists have demonstrated AdipoRon’s ability to
decrease insulin resistance, the abnormal distribution CHANGING BAD FAT TO GOOD
of adipocytes, and glucose intolerance in mice that have White adipose tissue (the type described earlier) is linked
been bred to be a model for obesity and type 2 diabetes. to obesity and insulin resistance. Brown adipose tissue,
Another adipokine, leptin, may sound familiar. This in contrast, regulates body temperature by burning
hormone promotes feelings of satiety. In other words, it calories to release heat. Unlike white fat, brown appears
tells the brain that you are not hungry. Myalept is a leptin to benefit us. In most adults, however, brown adipose
analog that has been approved by the FDA to treat the makes up only about five percent of total body fat.
rare disorder lipodystrophy, or the irregular distribution Scientists at the University of Pennsylvania
of adipose tissue within the body. Researchers at (Philadelphia, PA) have identified molecular signaling
Novelion Therapeutics (Vancouver, Canada) are studying pathways in white adipose tissue that converts them
the effect of Myalept in obese patients with low leptin to energy-burning brown fat. Learning to activate that
levels. Preliminary data suggests that people taking pathway pharmaceutically may eventually yield another
Myalept lost more weight than those taking a placebo. obesity-fighting drug.
Previous studies have suggested that Myalept may not
be useful in treating obesity. However, this research A team at Columbia University (New York City, NY))
didn’t specifically focus on people with very low natural has taken a more surgical approach to the problem.
levels of leptin. These encouraging results suggest that They’ve removed white adipose tissue from mice, treated
leptin deficiency may account for at least some types it in the lab to stimulate its conversion to brown fat,
of obesity. and then grafted it back onto the mice. Amazingly, the
“rehabilitated” tissue survived and functioned as brown
fat for the two-month duration of the experiment.
MITIGATING INFLAMMATION
And the Columbia research success goes beyond rodent
Inflammation appears to be a root cause of adipose
fat. They’ve also figured out how to convert human white
tissue dysfunction. Consequently, anti-inflammatory
fat into brown fat in the lab. However, they have not yet
medication presents an attractive option. A number of
carried out any grafting experiments in humans.
companies are testing approved or newly developed
therapeutics that take this approach: Taken together, these various approaches should yield
more effective treatments for obesity and insulin
resistance in the coming years.

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CRACKING THE CIRCADIAN CODE Life Science Training from Industry Experts

CRACKING THE CIRCADIAN CODE rhythms. Like the rest of our body, the digestive system
works on a 24-hour clock. Prime fat-burning starts six
The most recent WEEKLY described the molecular basis
to eight hours after our last meal and continues until we
of circadian rhythm, the internal 24-hour clock that
break the “fast.” Establishing a regular, time-restricted
governs many of our biological processes. Disrupting this
pattern of eating may optimize our digestion, nutrient
cycle over the long term is linked to diseases including
absorption, and fat-burning.
obesity, diabetes, heart disease—even cancer. Now we
turn our focus on some daily habits that can profoundly Eating on schedule may also promote a healthy gut
influence our internal clocks and therefore our health. microbiome. Researchers at the Weizmann Institute
of Science (Rehovot, Israel) have shown that the
TIME TO EAT microbiome composition of people who are jet-lagged
starts to resemble that of obese people. This alteration
To maintain a healthy weight, science tells us to watch
suggests that messing with circadian rhythms messes
how many calories we take in. Of course, it’s critical
with the microbiome of shift workers and travelers who
to think about the content of those calories too—
regularly change time zones. Further research from
unfortunately a half pound of hot fudge sundae does
Panda’s lab suggests that sticking to a time-restricted
not equal a half pound of chicken breast and steamed
eating schedule may offset this disruption.
broccoli. Interestingly, the latest research also suggests
that when we consume those calories matters too.
TIME TO EXERCISE
Scientists at the Salk Institute for Biological Studies (San
Nutrition and the timing of our meals constitute only
Diego, CA) took two groups of genetically identical mice
part of what’s needed for maintaining a healthy weight.
and gave them access to the same high-fat foods. Sadly
Exercise is another critical component. Could circadian
for one group though, they could enjoy the chow only
rhythms also influence the benefits of exercise?
during an eight-hour time period.
Studies suggest that exercising first thing in the morning
All of the rodents ate the same amount but the similarity
or in late afternoon best enhances its positive effects.
ends there. Like many of us in the U.S., the 24-hour
Early morning workouts take advantage of early morning
access group “grazed,” munching small amounts of food
daylight exposure (or bright lights at the gym). They
regularly. The other mice, meanwhile, ate fewer but
offer a direct way to synch the brain’s “suprachiasmatic
larger meals within the eight-hour window. Remarkably,
nucleus,” the master clock that governs the cells in the
the “eight-hour” mice didn’t gain excess weight or
rest of your body.
experience unhealthy blood glucose or blood cholesterol
levels. Their “24-hour” siblings did. Late afternoon exercise corresponds to the natural
circadian peak in muscle tone, or at-rest muscle
Follow-up studies also suggest benefit from limiting
contraction. Thus people may find high-intensity
when we eat to nine, ten, or even 12 hours. These longer
exercise or weight-training easier in the late afternoon
windows may prove easier for people to handle. The Salk
than at other times. In general, exercise physiologists
researchers think that the effort may be worth it. One
counsel against intense exercise close to bedtime
small study focused on a group of ten overweight people
because it raises levels of the stress hormone cortisol,
(BMI of greater than 25). They were told to consume
which can interfere with sleep.
all calories— including drinks and snacks!—within a
daily ten-hour window. Happily for them, they all lost a Moderate activity, such as walking after dinner, is
significant amount of weight after four months without another story. It may help regulate blood sugar levels.
changing the amount or type of food they ate. This is because insulin levels and glucose tolerance drop
nightly as part of a 24-hour cycle. Light to moderate
According to Dr. Satchin Panda, who is director of the lab
exercise also causes muscle cells to absorb glucose
involved, the participants’ success is linked to circadian

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from the blood stream, compensating for reduced that we work with our circadian clocks to obtain optimal
insulin levels. health. For even more information about this topic, Dr.
Regular sleep deprivation, whether from shift work, jet Satchin Panda’s new book, The Circadian Code, delves
setting or simply not taking care of ourselves can do more deeply into the science behind time-restricted
more than make us a little cranky—it can damage our eating and the interaction of our circadian rhythms and
health in serious ways. Maintaining a regular schedule our overall health.
for sleeping, eating, and exercise can help to ensure

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BREAKING DOWN DRUG METABOLISM Life Science Training from Industry Experts

DOWN THE HATCH! THEN WHAT? concentrations correspond directly with a decrease in
how well a drug works. So if you enjoy an occasional
When popping a pill, how often do we think about what
glass of grapefruit juice, double check your medication
happens next—to the pill, or to our bodies? Maybe we
labels. Examples of medicines that mix poorly or even
assume the body welcomes any extra help to soothe our
dangerously with the big citrus include:
headache or control our blood pressure. This WEEKLY
looks into the mystery of what comes after the swallow. • Statins, which help lower blood cholesterol levels,
and include medications such as Zocor (Merck;
Our bodies immediately respond to any pill (also known
Kenilworth, NJ) and Lipitor (Pfizer; New York City, NY)
as a small molecule drug) we ingest as an unwelcome
can result in increased levels via CYP3A4 inhibition.
intruder. It tries to get rid of the alien lickety split.
The process by which the body attempts to oust the • High blood pressure medications, such as Adalat
foreign chemicals forms the basis of drug metabolism. (Bayer Pharma; Berlin, Germany) may cause toxicity.
Understanding this process is a critical part of • The anti-depressant/anti-anxiety medicines Zoloft
drug development. (Pfizer) or BuSpar (Bristol-Myers Squibb; New York
The liver functions as the most important organ when it City, NY). Drug levels may go up.
comes to drug metabolism. So it makes sense to focus on • Erectile dysfunction medications such as Viagra
the liver enzymes that metabolize small molecule drugs (Pfizer) and Cialis (Eli Lily; Indianapolis, IN) may
and how they work. become toxic.
• Allergy medicines such as Benadryl ( Johnson &
ALL JUICED UP (OR DOWN) Johnson; New Brunswick, NJ) and Allegra (Sanofi;
Have you ever noticed the warning label on certain Paris, France) may potentially lose some of
prescriptions advising the patient to avoid grapefruit their effectiveness.
juice? It contains chemicals that can inhibit proteins
essential to drug metabolism. Depending on the protein, TYBOST: MAKING A PATHWAY
the beverage either increases the amount of the drug OUT OF A BYWAY
in a patient’s blood stream to potentially toxic levels or Inhibiting CYP3A4 is not always a bad thing. Gilead
decreases how much medicine reaches its target in the Science’s (Foster City, CA) Tybost is a drug that inhibits
body. Bad news either way. cytochrome P4503A enzymes, another group of drug-
Scenario 1: Too much drug.
 metabolizing liver proteins. The inhibition increases the
efficacy of certain antiviral drugs. Normally the enzymes
The small intestine and the liver are both home to one
break down these drugs. A combination of Tybost and
such protein, CYP3A4. Grapefruit juice inhibits CYP3A4,
the antivirals boosts the long-term concentrations of
which makes it accumulate in the patient’s bloodstream.
the antiviral medication, boosting their effectiveness.
These higher concentrations may cause direct toxicity
Tybost is approved both as a stand-alone drug and as a
or damage the liver over time. In general, higher than
component of a four-drug-combination anti-retroviral
prescribed amounts of a drug force the liver to work
therapy for HIV.
harder. Only medicines broken down by CYP3A4
enzymes are potentially affected by grapefruit juice. EASILY CONFUSED: PHARMACOKINETICS
Scenario 2: Not enough drug. VERSUS PHARMACODYNAMICS
Other medications use transporter proteins—proteins The field of pharmacokinetics (PK) focuses on how the
on cellular surfaces that allow molecules to enter cells. body affects a drug. In contrast, pharmacodynamics (PD)
Grapefruit juice inhibits these cellular gatekeepers, relates to how a drug affects the body.
which results in lower drug concentrations. Reduced

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Drug development includes studies, called ADME, to The continued sale of Plavix hinges on prospective
analyze a new product’s pharmacokinetics: patients taking a genetic test to see if they have the
• Absorption: the process of a drug entering anti-Plavix mutation. Doctors prescribe patients who
blood circulation. have the mutation another medicine that acts by a
• Distribution: the dispersion of the drug throughout different mechanism.
the body. GENOTYPING METABOLISM
• Metabolization: the body’s recognition and Two enzymes—CYP2D6 and CYP2C19—play a
subsequent breakdown of the parent compound into fundamental role in the metabolism of an estimated 25%
daughter metabolites. of all prescription drugs. CYP2C19 has three variants,
• Excretion: the elimination of the drug from the body. while the CYP2D6 gene has over 90 different variants.
Depending on a patient’s combination of variants, he
TERM OF THE WEEK: PRODRUG or she may metabolize the drug too quickly, too slowly,
Some drugs are designed to be prodrugs—given to or in just the right way. Roche Diagnostics (Basel,
patients in an inactive or somewhat inactive form. The Switzerland) markets a test that lets physicians quickly
patient’s own metabolism then fully activates the drug. determine a patient’s “metabolizer type” based on their
The anti-clotting medicine Plavix (Bristol-Myers Squibb) specific combination of genes. The test, AmpliChip
makes an excellent example of a prodrug. CYP450, uses SNP chip technology to assess a patient’s
When it first came to market, Plavix faced a recall. Nearly gene variants and ability to safely process certain
14% of patients on the medication suffered strokes prescription drugs.
or heart attacks due to blood clots anyway. Further Livers. They’re not all the same. Understanding the
investigation revealed patients with a mutation in the differences in how they handle prescription drugs is just
liver enzyme CYP2C19 couldn’t activate the Plavix, one more piece of the precision medicine puzzle.
making it ineffectual.

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SWALLOWING A BIOLOGIC DRUG? Life Science Training from Industry Experts

SURVIVING THE STOMACH IS KEY pH-sensitive and stays intact at the low pH of
the stomach.
Over the past two decades, biologic drugs — drugs
composed of proteins produced by living cells — have • When it enters the intestine, its outer coating
become the safest, most effective top sellers within the dissolves at the slightly higher pH of the intestine.
pharmaceutical industry. • This pH change also triggers a chemical reaction to
Approved to treat a variety of diseases including occur, which releases carbon dioxide which blows up
rheumatoid arthritis, diabetes, multiple sclerosis, a tiny “balloon” found within the pill.
Crohn’s disease, and a whole range of cancers, these • This supplies the force needed to drive the
drugs include monoclonal antibody therapeutics, drug-containing needle into the intestinal wall —
hormones, and immune system signaling molecules. delivering the biologic to the bloodstream.
Their safety and efficacy depend largely on the complex, • The needle itself then dissolves, and the balloon
three-dimensional structure of the protein product is excreted.
itself — which is incredibly delicate and time-consuming
This may sound like the stuff of science fiction — and
to develop on a large scale. The majority of these are
indeed, the device is still in preclinical trials — but the
administered via injection directly into the bloodstream
robotic pill has attracted big name pharma partners,
for maximum potency.
including AstraZeneca (London, UK) and Novartis
So, why can’t we just swallow a biologic pill? The simple (Basel, Switzerland), as well as investment from
answer: biologics would not survive the acidic pH and Google Ventures (Mountain View, CA) among other
digestive enzymes of the gut. Even if survival were venture funds. If the device ultimately succeeds, it will
possible, the next issue to contend with is absorption; revolutionize the delivery of biologic drugs.
if the protein is not broken down properly, it will not
be absorbed into the bloodstream. Companies seeking ENGENE’S GUT CELL FACTORIES
to develop oral delivery of biologics must overcome
enGene (Vancouver, Canada) is bypassing the delivery
both hurdles.
obstacle by attempting to turn the cells of the gut into
Injectable delivery is cumbersome at best. It has a drug-producing factories. The trick is to deliver a gene —
significant impact on a patient’s quality of life, which the instructions for a specific therapeutic protein — to
in turn affects compliance to drug therapy regimes. those cells. enGene is using tiny carbohydrate-based
In many cases, patients must make regular trips to an nanoparticles to encase the gene. The carbohydrate
infusion center to receive required treatments. Thus coating protects the gene as it passes through the
oral administration is the holy grail in terms of delivering stomach, yet allows it to be absorbed into the intestines.
biologics. In this WEEKLY, we’ll take a look at the different The protein is then made in the intestines.
paths being pursued to make this ambition a reality.
This approach is especially appealing for diseases
affecting the colon and small intestine, such as
THE ROBOTS OF RANI THERAPEUTICS
inflammatory bowel syndrome or Crohn’s disease.
One of the hottest companies in the oral delivery of enGene is currently using this platform in the preclinical
biologics space is Rani Therapeutics (San Jose, CA). development of the anti-inflammatory protein IL-10, for
Their product is a “robotic pill” — a small device capable the treatment of inflammatory bowel disease.
of traversing the intestinal tract and injecting a biologic
drug directly into the body. It works in the following way: APPLIED MOLECULAR TRANSPORT
• After the pill is swallowed, it makes its way through Applied Molecular Transport South San Francisco,
the digestive process, keeping its fragile biologic CA) is using a protein scaffold adapted from pathogenic
drug cargo tucked safely inside. Its capsule is

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microbes such as salmonella, which colonize the gut by Swallowing biologic drugs is no longer a pipe dream.
secreting immune-crippling proteins into our body. Sensing the opportunity to make a significant impact
Proteins from these “gut bugs” work by tricking the on the biopharma industry, innovative companies are
intestines into absorbing toxic proteins in the same way approaching the drug delivery problem with a range of
that they absorb nutrients from food. Applied Molecular strategies. It is likely that no single strategy will work
Transport scientists have tweaked these microbial for all types of biologics, but any success with a handful
proteins to carry a therapeutic payload, rather than of products would represent a major breakthrough for
the toxins. Development efforts are currently aimed at the industry.
delivering anti-inflammatory proteins to the intestines.

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TAKING A SWING AT ALLERGIES Life Science Training from Industry Experts

HOW DO ALLERGIES DEVELOP? TERM OF THE WEEK: HISTAMINES

Watching a game at the ballpark and digging into a Histamines are small molecules produced by mast cells.
bag of peanuts is a source of entertainment for many Once released, histamines bind to receptors on the
Americans. For the 15 million who suffer from peanut surface of blood vessels, increasing their permeability —
allergies, the idea of being taken out to the ballgame the ease with which fluid moves out of the blood vessels.
elicits concern — or even anxiety. Histamines also bind to receptors on certain types of
Food allergies — think tree nuts, milk, eggs, wheat, nerve cells, resulting in muscle contraction. A host of
soy, fish, and shellfish — are on the rise. The mere dust symptoms can be triggered, ranging from annoying to
particle of a freshly cracked peanut can be responsible deadly. Typical signs of histamine release include:
for an unpredictable cascade of reactions, including • Increased blood vessel permeability resulting in a
death brought about by anaphylaxis. runny nose and watery eyes.
This WEEKLY takes a swing at explaining how allergies • Increased muscle contraction leading to throat
develop, the current treatments, and what new products constriction and difficulty breathing.
might change the way allergen desensitization therapy • Extreme fluid release from tissues causing a sudden
is delivered. drop in blood pressure, potentially bringing on a
heart attack.
SOMETHING TO SNEEZE AT
• Difficulty breathing and swallowing, swelling, heart
The host of symptoms dubbed “allergies” are the
palpitations, and unconsciousness — sometimes
end result of the immune system’s response to a
causing death.
normally harmless substance, as if that harmless
substance were a threat. An initial allergen exposure Mild allergy symptoms such as runny nose and watery
results in the production of a class of antibodies called eyes can often be successfully controlled by the use of
Immunoglobulin E (IgE). A second exposure to the an over-the-counter antihistamine, a drug that works by
allergen results in an “allergen-IgE antibody complex” blocking the interaction of histamines with receptors on
These newly produced complexes bind to and activate nerve and muscle cells. However, once anaphylaxis — a
mast cells — a type of immune cell. As the image below severe allergic reaction that includes difficulty breathing
shows, activated mast cells send out chemical alarms in and heart palpitations — has occurred, it is too late for
the form of histamine. antihistamines to be effective. These symptoms can only
be treated with an injection of the hormone epinephrine
— and the sooner, the better in cases where a life is on
the line.
Epinephrine helps to reverse histamine’s effects by
decreasing blood vessel permeability, relaxing muscle
cells, and stimulating the heart. People at risk for
anaphylaxis need access to an epinephrine auto-injector
— a spring-loaded syringe that makes the lifesaving shot
readily available. This type of product is referred to as
a “combination product” because it combines a device
(the auto-injector) with a medicine (epinephrine). Mylan
(Canonsburg, PA) EpiPen is an epinephrine auto-injector,
as is Amedra’s (Horsham, PA) Adrenaclick.

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THE HYGIENE HYPOTHESIS allergy was assessed – and an 81 percent reduction
in allergy was found in the children who regularly
Epidemiologists have noticed an interesting trend as
consumed peanuts when compared to those who
countries rise from developing to developed status:
avoided them
an improvement in sanitation and access to antibiotics
means less pathogenic exposure and lower infection ALLERGENS BY THE DOSE
rate. As the infection rate drops, the incidence of
The current allergen immunotherapy market includes
allergies shoots up.
allergy shots (which require monitoring by a physician)
Many scientists think early exposure to infection helps and drops or tablets dissolved under the tongue
shift the immune response towards fighting pathogens (which can sometimes be taken at home). Aimmune
while minimizing the production of IgE antibodies. Therapeutics’ (Brisbane, CA) AR 101 is a dissolvable
Exposure to potential allergens (while the immune tablet made of pharmaceutical grade peanut protein that
system is still developing) helps to desensitize the can be mixed with food as a means of delivery.
allergic response.
AR 101 Phase II studies report patients becoming
THE STRATEGY BEHIND desensitized to doses at least twenty times greater
DESENSITIZATION THERAPY than the original allergy-inducing dose — and in some
cases, more than 100-fold greater. The goal is to reach a
The idea of allergy desensitization through controlled
tolerance level that offers protection against accidental
exposure has been around for decades. Desensitization
eating of peanuts. The product was awarded Fast Track
is the principle behind allergy shots shown to be
status by the FDA and is now in Phase III. Aimmune has
effective against pet dander, dust mites, and pollen.
plans for clinical studies on immune system training for
Desensitization therapy was once considered to be too
egg and milk allergies to begin later this year.
risky for food allergies, but a number of new studies
support the idea that gradual exposure to food allergens DBV Technologies (Bagneux, France) is also in the
may be beneficial. allergy fight. They are currently conducting clinical trials
on their Viaskin skin patch which delivers low doses of
In January 2017, the National Institutes of Health
either peanut, milk, or dust mite allergens. By delivering
(NIH) released new guidelines recommending the early
an allergen through the skin instead of the blood,
introduction of peanuts into children’s diets, including
the body will react less severely, reducing the risk of
those considered to be at high risk for developing peanut
anaphylaxis as a side effect. The Viaskin peanut patch,
allergies because they already have severe eczema or
currently in Phase III clinical testing, has been awarded
egg allergies. These new guidelines are based on the
Fast Track designation by the FDA.
results of the NIH-funded Learning Early About Peanut
Allergy (LEAP), a study which randomly assigned 600 As allergen desensitization treatments continue to make
high-risk infants to either peanut-avoidance or the their way through the drug pipeline, allergy sufferers
regular inclusion of small amounts of peanut products in remain hopeful for better and easier treatment options,
their diet for the first five years of life. At age five, peanut and maybe even one day — a cure.

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NEW HOPE FOR SPINAL MUSCULAR ATROPHY Life Science Training from Industry Experts

FIRST THERAPY APPROVED FOR SMA is non-functional and degrades shortly after being
produced. Patients with less severe forms of the disease
Squeaking by on December 23rd as the last new drug
usually have extra SMN2 copies because ultimately,
approval of 2016, Biogen’s (Cambridge, MA) Spinraza
even tiny amounts of SMN protein provides some motor
now provides hope for the thousands of families
nerve function.
affected by a debilitating neuromuscular disorder known
as spinal muscular atrophy (SMA). SMA robs people of The four generally accepted classifications of SMA are:
their ability to walk, eat, and ultimately, breathe. • Type 1: The most severe and the most common.
In addition to Spinraza, there are 13 other new therapies Babies do not move, but lay perfectly still in their
making their way through the clinic, according to the cribs. As the disease progresses, toddlers have
patient advocacy group Cure SMA. The increase is trouble with swallowing and respiratory function.
largely due to a better understanding of the disease SMA Type 1 is usually fatal by age two.
and a surge in funding for basic and clinical research. • Type 2: Symptoms manifest between six and
SMA affects about 1 in 10,000 babies born in the eighteen months. These children can typically sit
United States. but not stand or walk. Respiratory function is often
In this weekly, we’ll decipher the science behind SMA, compromised, however with the help of machines
explain the novel mechanism of action used by Biogen’s many of these patients live into adulthood.
new drug, and find out how other drugs in development • Type 3: Symptoms occur after age one. These kids
are zeroing in on this genetic disease. are usually able to walk, but may lose that ability as
the disease progresses. Respiratory function is less
SMA PRIMER impaired, and life expectancy is often near normal.
Our nervous system consists of the brain, spinal cord, • Type 4: This is the adult-onset form, typically
and a vast network of nerves that feed into every tissue developing at age 30 or later. Muscles gradually
of the body. Motor neurons are a type of nerve cell weaken, and the patient often needs to use
that sends messages from the spinal cord to muscles, a wheelchair later in life. Life expectancy is
enabling movement. not affected.
In order for the motor neurons to do their job, a SMA Type 1 is the most common and most severe,
functional protein called the survival motor neural (SMN) making up 60% of cases. As a result, many companies
protein is necessary. The survival motor neuron 1 (SMN1) are looking to tackle this segment of the disease
gene is responsible for producing most of the SMN population. Below find a few treatment approaches for
protein used by the body. A second, closely related SMA Type 1.
gene is the survival motor neuron 2 (SMN2) gene, which
produces a much smaller amount of SMN protein and is ON THE MARKET: ANTISENSE THERAPY
seen as a sort of “back-up” version to SMN1. Developed by Biogen in partnership with Ionis
SMA is caused by a variety of mutations in the SMN1 Pharmaceuticals (Carlsbad, CA), Spinraza is one of a
gene. Without functional SMN protein, the neurons do small but growing class of drugs: antisense therapeutics.
not work correctly and eventually die. How soon they It is a synthetic mRNA molecule that binds to the
die depends on the extent of the SMN deficiency, which naturally occurring SMN2 mRNA in such a way that more
correlates with the severity of the disease: the less SMN of the mRNA is used to make the protein. The result is
produced, the more severe the disease. greater amounts of full-length, functional SMN protein.
The back-up gene, SMN2, produces a small amount of Recall from high school biology that mRNA provides the
functional SMN protein. However, differences in the way instructions to make proteins. If the mRNA is working
SMN2 functions means most (but not all) of the protein properly, the correct, functional protein is made.

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IN DEVELOPMENT: SMALL that has been stripped of its disease-causing ability,
MOLECULE ENHANCERS and modified to carry a correct version of the mutated
gene. The viral vector can also be engineered to drop its
PTC Therapeutics (South Plainfield, NJ) — in partnership
genetic cargo into specific cells. In the case of SMA Type
with Roche (Basel, Switzerland) — has begun Phase
1, the AAV9 vector crosses the blood-brain barrier and
II clinical studies on its proprietary small molecule
delivers corrected copies of the SMN1 gene into motor
drug, RG67800. RG67800 is similar to Spinraza in that it
neuron cells in the brain.
changes the way nerve cells process the SMN2 mRNA,
resulting in increased production of functional SMN AveXis (Bannockburn, IL) has a gene therapy candidate,
protein. However, a notable difference is the delivery AVXS-101, in Phase I/II clinical studies. The company
mechanism—Spinraza is an injectable while RG67800 reported promising results in mid-2016, noting that
is a pill. Novartis (Basel, Switzerland) is preparing for the drug appears to be safe and effective. Babies
Phase I clinical testing of a small molecule modulator of who received this gene therapy showed marked
SMN2 mRNA. increases in SMN production and in movement. Also
in the gene therapy mix is Voyager Therapeutics
IN DEVELOPMENT: A GENE (Cambridge, MA) with their treatment currently in
THERAPY CURE? preclinical development.

As a single gene disorder, SMA is an ideal candidate for a With additional approvals hopefully to follow in the wake
gene therapy approach. Scientists deliver the corrected of Spinraza, SMA patients and their family members are
version of the mutated gene by a viral vector — a virus hoping to kiss this devastating illness goodbye.

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GMO APPLES HIT MARKETPLACE Life Science Training from Industry Experts

COMING SOON TO A Recall that RNA serves as the intermediary between DNA
SUPERMARKET NEAR YOU and proteins. The information in DNA is converted to
RNA, which is then translated into a protein by organelles
Coming soon to a supermarket near you: apples that
called ribosomes. One way to stop PPO production is to
have been genetically modified to resist browning.
get rid of the PPO RNA before it enters the ribosome.
Dubbed Arctic Apples, the plants were approved two
Enter RNAi.
years ago by the USDA and will begin appearing on select
retailers’ shelves in Midwestern U.S. states this month. RNAi is a short, complementary RNA sequence that
attaches to a target RNA, in this case the target is PPO
Developed by Okanagan Specialty Fruits (Summerland,
RNA. This creates a double-stranded RNA, which is
B.C., Canada), the Arctic Apple also represents a new
universally recognized as viral RNA and is destroyed
type of genetically altered food — one that has been
by another enzyme known as RNA-induced silencing
engineered to directly appeal to and benefit the
complex (RISC). In the case of the Arctic Apple, RNAi
consumer rather than the farmer. Their debut may also
attaches to the PPO RNA and is destroyed by RISC. No
help ease lingering safety concerns voiced by GMO critics
PPO RNA ever makes it to the ribosome, so no PPO is
due to the innovative technology used to create Arctic
ever produced by the apple. No enzyme, no chemical
Apples. Let’s take a look at what causes apple browning,
reaction, no browning.
how science is able to produce a remedy, and preview
what’s on the horizon for consumer-oriented genetically WHY LESS BROWN?
modified food.
Why go to all this trouble to make apples less brown?
OXIDATION CAUSATION This modification is expected to benefit growers,
packers, shippers, and retailers by limiting the amount
The browning observed in conventional apples is the end
of bruising and other visual imperfections in their
result of an oxidation reaction — a chemical reaction
apples — meaning less waste and more product to sell.
between oxygen and another substance. Typically,
Food processors, likewise, will be able to produce more
oxidation reactions result in a color change (think rusting
consistent juices, sauces, and sliced apple products,
car — the oxidation of metal). In apples, browning is the
without relying on antioxidant treatments currently
oxidation of “phenolic compound” by an enzyme called
in use.
polyphenol oxidase (PPO).
And since Arctic Apples contain no foreign DNA, they
In order for oxidation to occur, PPO has to come into
are likely to be more palatable to GMO-wary consumers.
contact with the phenolic compounds. Within the apple
The apples have undergone 10 years of field testing,
cells, phenolic compounds are typically sequestered
and do not differ in any significant respect from non-
inside of membrane-bound compartments. If the cell
modified apples, apart from the lack of PPO enzyme.
is disrupted, by slicing or dropping the apple, the seal
Based on field studies, blocking the production of PPO
of these compartments is broken and the phenolic
does not make apple trees more susceptible to pests,
compounds exposed. PPO is then able to act, oxidizing
and so their adoption should not result in any increased
the compounds to create browning.
pesticide use.

SILENCING THE GENE ON THE MARKET

Scientists at Okanagan Specialty Fruits chose to tackle
The first two varieties of modified Arctic Apples
apple browning by blocking the production of the PPO
appearing on the market are Golden Delicious and
enzyme. No enzyme, no chemical reaction, no browning.
Granny Smith. If successful, they could pave the way
They used a gene-silencing technique known as RNA
for other consumer-oriented products such as non-
interference (RNAi) to block the production of PPO.

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browning cherries and pears, also in development at to knock out a second protein, asparagine synthetase-1.
Okanagan Specialty Fruits. When potatoes are cooked at high temperatures,
A similar product — the Innate Potato — was approved in asparagine synthetase-1 reacts with the potato sugars
November 2014 by the USDA. Developed by J.R. Simplot to produce a chemical called acrylamide — which has
(Boise, Idaho), Innate also uses RNAi technology to been linked to cancer in rodents. The Innate potatoes are
decrease production of the PPO protein, again with the already on the market in the U.S. at select retailers.
goal of reduced browning. Innate potatoes also use RNAi

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STEM CELL SNAPSHOT Life Science Training from Industry Experts

INDUCING STEM CELLS TO HEAL Let’s take a look at companies trying to boost the
regenerative capacity of these tissues less adept at self-
Headlines touting stem cells often claim the therapies
repair.
heal everything from hair loss to hearing loss. While
many of these treatments are not FDA approved, there PUSHING THE PROGENITOR
are some promising innovations winding through
Progenitor Therapeutics’ (Stevenage, UK) name hints to
preclinical and clinical development. Here at WEEKLY
the company’s main technology. A progenitor cell is one
headquarters, we like to tease out the science behind
that has taken the first steps to becoming a specialized
the scene, so let’s review regenerative medicine basics
cell, such as a cardiac cell. Once these steps are taken,
and survey the companies attempting to repair damaged
the stem cell is “committed” to becoming that specialized
tissue with these high potential cells.
cell type.
STEM CELL PRIMER Progenitor Therapeutics is focused on discovering small
Stem cells are unspecialized cells that have the ability molecule drugs that will nudge adult stem cells down
to develop (differentiate) into 1 of 200 cell types in the the differentiation pathway. They do this by testing
body. There are two general classifications: hundreds of thousands of drugs on stem cells to see
which bring out progenitor cell types. Although still in
• Embryonic stem cells, found only in developing
preclinical development, their work offers a tantalizing
embryos, can become any cell type within the
future of popping a pill to regenerate damaged tissue.
adult body.
• Adult stem cells, found within organs of a fully IGNITING NEURAL STEM CELLS
developed body, can become only certain cell types. Neuronascent (Clarksville, MD) and Neuralstem
Typically these cell types come from the organ in (Germantown, MD) are leading the charge in developing
which they are found. small-molecule activators of neurogenesis, or the
Embryonic stem cells have the most therapeutic generation of neurons (brain cells) from stem cells.
potential. A main focus of research is sussing out which By screening large chemical libraries, scientists at
combo of hormones cause a stem cell to commit to these companies have identified compounds that
becoming a specific tissue; for example, spinal cord activate neurogenesis from adult neural stem cells
tissue vs. brain tissue. The billion dollar market: grow both in the lab and in mouse models of various
and transplant stem cells into patients who have lost neurodegenerative disorders.
tissue due to acute trauma. Neuralstem’s lead neurogenesis candidate, NSI-189,
Since embryonic stem cells may come from unrelated increased the size of the hippocampal region of mice
donors, there is the likelihood of tissue rejection. Thus brains by 20%. Neuralstem is beginning Phase II of NSI-
there is considerable interest in developing therapies 189 for major depressive disorder. The drug is already in
using a patient’s own adult stem cells. preclinical development for Alzheimer’s disease.
In certain tissues such as bone marrow, muscle, liver, Neuronascent’s first focus is the development of small
and skin, damaged or worn out areas are naturally molecule compounds that promote neurogenesis for
and regularly replaced by activated adult stem cells. In the treatment of Alzheimer’s disease. In an Alzheimer’s
other tissues such as heart and brain/spinal cord, adult mouse model, the lead compound NNI-362 promoted
stem cells exist in such small numbers, our body cannot the growth of new hippocampal neurons that not only
readily activate them to replace tissue. Enter stem migrated to the correct functional location, but also
cell therapy. differentiated and survived long enough to reverse
previously observed cognitive declines. Neuronascent is
preparing for Phase I of NNI-362.

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ON THE FAST TRACK
WITH ANGIONETICS
Last month, Angionetics (San Diego, CA) won fast track
designation for its gene therapy candidate, Generx,
which is currently in Phase III clinical studies.
Generx is a viral vector — a virus that has been modified
to deliver a specific gene to a target cell type — that
delivers a gene for fibroblast growth factor 4 (FGF4) to
cardiac tissue damaged due to lack of oxygen during
a heart attack. The damaged tissue then produces
the FGF4 protein, which promotes the formation of
new blood vessels in damaged regions, increasing the HEAR, HEAR
heart’s ability to deliver oxygen-rich blood. The FGF4
Good hearing depends on thousands of tiny hair cells
gene delivered by Generx does not incorporate into the
within the inner ear that mechanically amplify low-
cardiac cell’s genome. This means the growth factor is
level sound. As these hair cells are damaged or lost
only produced for a few weeks — just long enough to
through exposure to excessive noise, toxic drugs, and
jump start the growth of new blood vessels.
normal aging, hearing loss occurs and we are not able to
generate new hair cells.
That may change. The leaders behind Cambridge-based
startup Frequency Therapeutics reported the discovery
of a small-molecule activator of hair cell precursors —
LGR5+ cells — which are a type of adult stem cell. Once
activated, LGR5+ cells differentiate into hair cells. The
work was done on animal models in the lab using cochlea
– the portion of the inner ear which contains the hair
cells. The findings suggest Frequency may be on the path
to developing a drug to restore hearing loss.

WANTING THE WNT PATHWAY

NAKED DNA
Just last month, a new regenerative medicine company,
Juventas (Cleveland, OH) is also developing a gene Surrozen, was launched in South San Francisco.
therapy for heart disease. However, instead of using The company founders include scientists who have
modified virus, Juventas relies on plasmids — small, spent decades deciphering a cell signaling pathway
circular pieces of DNA engineered to contain the known as the Wnt pathway, which plays a central role
therapeutic gene. In this case, the plasmids are injected in stem cell maintenance and tissue regeneration.
directly into damaged cardiac tissue. Advantages to this Surrozen’s focus is on developing drugs that activate
“naked DNA” approach include reduced patient immune this pathway, potentially leading to treatments for
response and lower manufacturing cost. degenerative diseases.
Dubbed JVS-100, the Juventas plasmid contains a If any of these companies are successful a shot of stem
gene for stromal-cell derived factor-1 (SDF-1), which cells — not major surgery — may repair previously
recruits cardiac stem cells and promotes new blood unfixable tissue damage. We’ll keep a close eye on these
vessel growth. Like Generx, JVS-100 does not integrate exciting developments and be sure to report them back
into the recipient’s own DNA, making the production to you.
of SDF-1 temporary. JVS-100 has completed Phase I
clinical studies.

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THERAPEUTIC ANTIBODY PRIMER Life Science Training from Industry Experts

BASICS & INNOVATIONS cancer cell. When injected into a patient’s body, this
binding triggers the patient’s immune system to attack
Monoclonal antibody (mAb) therapeutics burst onto the
the cancer.
healthcare scene 20 years ago, and today they remain
one of the most versatile and effective therapeutics EASILY CONFUSED: MONOCLONAL
available. The tried and true mAbs are still in high VS. POLYCLONAL ANTIBODIES
demand, and we suspect this first wave of derivative
Commercially available antibodies are divided into two
products clamoring their way through the pipeline will
categories: Monoclonal and polyclonal. Monoclonal
be equally as successful. In this WEEKLY, we’ll review the
antibodies are antibodies that are all produced by the
basics of monoclonal antibodies and highlight some of
same B-cell, or by the clones (descendants) of that B-cell.
the recent innovations within this therapeutic modality.
This means they all recognize the same epitope on a
MAB PRIMER target immunogen. Therapeutic antibodies are always
monoclonal — this is required to ensure a consistent
Antibodies are proteins produced by B-cells, a type of
therapeutic effect.
white blood cell, in the immune system. Each antibody
has a distinct shape that recognizes a unique target, Polyclonal antibodies are produced by a collection of
often that unique target is a protein on the surface different B-cells, and recognize multiple epitopes on
of a pathogen. These foreign proteins are known as the same immunogen. Polyclonal antibodies are used in
immunogens. When an antibody recognizes its specific diagnostic and research applications, where detection of
immunogen, it physically binds to it. This binding action a specific immunogen is the only requirement, and the
alerts the immune system to attack and eliminate specific epitope recognized doesn’t matter.
the pathogen.
A BISPECIFIC CONNECTION
The specific protein that triggers antibody recognition
Antibodies are Y-shaped proteins. In antibodies
is called an immunogen, and the exact part of the
produced by our own immune system, as well as most
immunogen that is recognized by the antibody is called
therapeutic antibodies, the two “arms” of the Y are
an epitope.
identical, meaning that each antibody recognizes only
one target. Bispecific antibodies, in contrast, have been
genetically engineered by splitting and fusing the genes
for two different monoclonal antibodies together to
make a new Y. This way, the bispecific antibody is able
to recognize two different targets and bring them in
contact with one another.
For example, one arm might recognize a cancer cell,
while the other arm might recognize and bind to a killer
T-cell — a type of white blood cell that has the ability
to inject toxins directly into the cancer cell. By bringing
a cancer cell into direct contact with a killer T-cell,
This ability of antibodies to recognize and bind the T-cell is activated to kill that cancer target. This is
to a specific target is what makes them effective the mechanism of action used by Blincyto (Amgen;
therapeutics. And researchers have expanded on this Thousand Oaks, CA), the first and so far only bispecific
concept — for example, scientists may identify an antibody approved by the FDA.
antibody that binds to a protein on the surface of a

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 Currently, there are two ADCs on the market: Seattle
THE ABCS OF ADCS Genetics’ (Seattle, WA) Adcetris for the treatment of
Antibody-drug conjugates (ADCs) are highly potent, Hodgkin’s lymphoma, and Roche’s (Basel, Switzerland)
targeted therapeutics that work by combining the Kadcyla for the treatment of HER2-positive breast
targeting capabilities of monoclonal antibodies with the cancer. There are dozens more in clinical development
by companies including AstraZeneca (Cambridge, UK),
cancer-killing ability of toxic drugs, enabling the killing of
Roche, Immunogen (Waltham, MA), Novartis (Basel,
cancer cells with less impact on healthy cells. ADCs have
Switzerland), and Fortis Therapeutics (San Diego, CA).
three key components:
• A monoclonal antibody that is highly specific for a PHOTOIMMUNOTHERAPY
tumor-associated immunogen that has little to no DELIVERS A PUNCH
expression on healthy cells.
Photoimmunotherapy delivers the latest twist on
• A small molecule drug that is highly toxic and antibodies. Developed by the National Institutes of
designed to kill the cancer cell once internalized. Health, this technology uses light to deliver a punch.
• A chemical linker that connects the small molecule • A light sensitive, non-toxic drug is attached to the
drug to the antibody. The linker is stable in blood end of an antibody. This drug becomes toxic when
circulation but releases the toxin once inside exposed to infrared light.
the tumor.
• Infrared light is shone, activating the drug and
How does it work? damaging the cancer cell membrane. This damaged
• The antibody binds to its target immunogen on the membrane allows water to enter, causing the cancer
surface of the cancer cell. cell to burst and die.

• The antibody-drug conjugate is then taken up or • Upon bursting, the cancer cell releases tumor
internalized by the cancer cell. immunogens that serve to activate the immune system
to further recognize and target more tumor cells.
• Once inside the cancer cell, the linker is degraded
and the active drug released. One caveat to using photoimmunotherapy is the
tumor must be accessible to an infrared laser in
The ability to target only cancer cells allows drug
order to activate the photosensitive drug. Aspyrian
designers to use drugs that are much more toxic
Therapeutics (San Diego, CA) is in Phase II clinical
than traditional chemotherapy. The ADC has a higher
studies of photoimmunotherapy antibodies that target
specificity and only attacks cancer cells, avoiding
head and neck cancer.
nearby healthy tissue which is often destroyed
by chemotherapy. Over the years, monoclonal antibodies have proven
to be safe and effective therapeutics in a number of
different indications, most notably various cancers
and autoimmune disorders. Innovations in antibody
technology ensure monoclonal antibodies will
remain one of the most powerful tools in the drug
development arsenal.

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THE MECHANICS OF MELANOMA Life Science Training from Industry Experts

SPECTRUM OF THERAPIES GENETIC FACTORS: P53 & BRAF

Melanoma accounts for less than 1% of skin cancer Although most cases of skin cancer are traceable to
cases yet causes the vast majority of skin cancer deaths. excessive UV radiation from sun exposure, about
If detected early enough, melanoma is almost always 10% are likely due to genetic factors. The gene most
curable. If not, its ability to metastasize makes it difficult commonly mutated in familial melanoma is p53. p53 is
to treat. a “tumor suppressor” — it detects DNA damage in cells
Melanoma is more common in young adults than many and triggers either DNA repair pathways or activates
other types of cancer, with 25% of new cases occurring cell death if the damage cannot be repaired. Another
in people under age 45. Its prevalence is growing — gene, known as the BRAF gene, regulates cell growth
the number of new cases per year relative to the total and is mutated in inherited forms of melanoma. About
population has doubled since 1973. In 2017, there will half of all genetically-based melanomas have the
be an estimated 87,110 new cases of melanoma in the BRAF mutation.
U.S. and 9,730 melanoma-related deaths, according to Let’s take a closer look at BRAF. BRAF codes for a protein
the Aim at Melanoma Foundation. Let’s review the required for the transmission of a growth signal from a
basics and find out the latest treatments in the battle cell surface receptor to the cell nucleus (growth signal
against melanoma. transduction). Growth signaling is initiated by a growth
factor binding to its receptor. This binding transmits
MELANOMA’S METHOD a signal through the membrane, causing the internal
Melanoma is the uncontrolled growth of melanocytes, portion of the receptor to interact with and activate
the pigment-producing cells located in the bottom layer a protein inside of the cell. This activation is then
of the skin’s outermost layer (the epidermis). In skin transferred to the next protein in the pathway, and so on
cancer, a tumor initially grows and spreads within the until the signal reaches the last protein in the pathway.
epidermis due to DNA damage, which is usually caused When this protein is activated, it enters the nucleus,
by ultraviolet (UV) radiation. where it turns on specific genes that make proteins
which initiate cell division. BRAF is one of the proteins
If melanoma is detected during the epidermal level
in this pathway. In BRAF-associated melanoma, the
growth stage, it can often be surgically removed.
mutated BRAF is always turned on even when no growth
Penetrating the deeper layers of the skin as it grows,
factor is present.
it will eventually come into contact with lymph and
blood vessels — which act as a cancer highway. When
melanoma spreads to other parts of the body it is known
as metastatic melanoma.
Lighter-skinned people are at higher risk for melanoma
because the increased skin pigmentation found in darker
skin tones helps to block UV rays from penetrating and
damaging skin cell DNA. However, darker-skinned people
can and do get skin cancer. Another general risk factor
for melanoma is atypical moles, or moles with irregular
shape, color, or size. Moles are clusters of melanocytes
and so sudden changes in them may be an early warning
sign of melanoma. The small molecule drugs Zelboraf (Genentech; South
San Francisco, CA) and Tafinlar (Novartis; Basel,

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Switzerland) inhibit overactive BRAF and are approved INCYTE’S INSIGHT
for the treatment of late-stage melanoma.
Incyte (Alapocas, Delaware) has a new type of small
IMMUNOTHERAPIES IN THE FIGHT molecule immunotherapy in Phase III called Epacadostat
– it inhibits the enzyme IDO1. IDO1 helps regulatory
A few different checkpoint inhibitor therapies are on
T-cells to develop and become activated. Regulatory
the market for metastatic melanoma. These therapies
T-cells suppress the immune response, and therefore
enable killer T-cells – immune system cells that recognize
help cancer cells to escape immune surveillance.
and kill threats such as cancer cells – to become fully
Inhibiting IDO1 should suppress the development of
activated and able to kill tumor cells. Keytruda (Merck,
regulatory T-cells, bolstering the immune response
Kenilworth, NJ) and Opdivo (Bristol-Myers Squibb, New
against melanoma.
York, NY) both target PD-1, an inhibitory protein on the
surface of T-cells; Yervoy (Bristol-Myers Squibb) targets THE IMPLICATIONS OF MICRORNA
a second inhibitory protein, CTLA-4. Both PD-1 and
Researchers at Tel Aviv University examined how
CTLA4 essentially act as “off switches” for killer T-cells.
melanoma metastasizes in an article last summer:
By inhibiting these off switches, the killer T-cells become
the melanoma cells release tiny vesicles containing
fully activated, and able to kill melanoma cells.
microRNA, a type of regulatory RNA produced by all cells.
A second type of immunotherapy approved for
The microRNA-filled vesicles induce changes in the
melanoma is Amgen’s (Thousand Oaks, CA) oncolytic
layer of skin just below the epidermis, where melanoma
virus therapy, talimogene laherparepvec (T-VEC). An
originates. That lower layer of skin, known as the dermis,
oncolytic virus is a virus that infects and kills cancer cells.
contains blood vessels which cancer cells are able to
The cancer cells are killed through cell lysis: as the virus
access. The Tel Aviv team is identifying drug candidates
multiplies inside of the cells, it causes them to burst
that may interfere with this process, preventing the
open. This in turn releases new infectious particles that
metastasis that makes melanoma so deadly.
can target remaining tumor cells. In addition to killing
cancer via lysis, the presence of an actively replicating Increased understanding of the molecular pathways
virus triggers the immune response to target the tumor. that contribute to melanoma’s development and
T-VEC was designed to infect and kill melanoma cells, and spread will provide additional tools to fight those cases
is so far the only FDA-approved oncolytic virus. of metastatic melanoma that inevitably will continue
to arise.

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THE INTRIGUE OF HIF Life Science Training from Industry Experts

THE VERSATILITY OF HIFS response to reduced circulating red blood cells, leading
to anemia.
Quite a few headlines touting a term called “HIF
compound” or “hypoxia-inducible factor compound” Erythropoietin production can be enhanced by
have intrigued us here at WEEKLY headquarters. increasing the amount of HIF present.
GlaxoSmithKline (London, England), Akebia Under normal oxygen conditions, small amounts of
Therapeutics (Cambridge, MA) and more have HIF- HIF are produced, but are quickly degraded though the
inducing drugs in the pipeline which may prove to be action of an enzyme called HIF prolyl-hydroxylase (HIF
attractive alternatives to Amgen’s (Thousand Oaks, CA) PHD). HIF PHD is inhibited in low-oxygen conditions,
injectable Epogen currently on the market. In earlier enabling HIF levels to increase. Several companies are
stages of research, HIFs are being studied for their developing small molecule, orally available drugs to
connection to tumor growth. How can one compound inhibit HIF PHD, facilitating the activation of HIF under
be versatile enough to affect both anemia and cancer? normal oxygen levels. HIF PHD inhibitors in clinical
Without further ado, let’s take a look at the story behind development include:
hypoxia-inducible factors. • Akebia’s vadadustat, Phase III clinical development
for CKD anemia
TERM OF THE WEEK: HYPOXIA-
INDUCIBLE FACTOR • FibroGen’s (San Francisco, CA) roxadustat, Phase III
clinical development for CKD anemia
Hypoxia occurs when the amount of oxygen reaching a
person’s cells and tissues is inadequate. Hypoxia may • GlaxoSmithKline’s daprodustat, Phase III clinical
be triggered by lower oxygen concentrations at higher development for CKD anemia
altitudes or by disease processes seen in pulmonary
disorders, anemia, or circulatory deficiencies.
DISRUPTING ANGIOGENESIS IN CANCER
HIF is also thought to contribute to the process of
The low-oxygen environment in hypoxia causes cells to
angiogenesis — the growth of blood vessels into tumors.
make a protein called hypoxia-inducible factor (HIF). HIF
Most solid tumors have a hypoxic environment, due to
is a transcription factor — a protein that binds to cellular
their high cell density and lack of supporting vascular
DNA in a defined location and “turns on” specific genes
networks. This hypoxic environment causes cancer cells
(which then make their intended proteins).
to produce HIF, which in turn activates the secretion
HIF activates genes involved in the production of oxygen- of vascular endothelial cell growth factor (VEGF). VEGF
carrying red blood cells and the formation of blood triggers angiogenesis, which provides a way for tumors
vessels (angiogenesis). Both of these processes assist in to get oxygen and nutrients, enabling the tumor to
increasing oxygen delivery to hypoxic (oxygen-deprived) continue growing. Angiogenesis also provides a possible
tissues. route for individual tumor cells to exit the tumor and
spread to other parts of the body.
ACTIVATING HIF FOR ANEMIA
Because of this relationship between HIF, angiogenesis,
Anemia is the decrease in the total amount of red blood
and cancer, there is considerable interest in developing
cells in the body, resulting in a lowered ability of the
HIF inhibitors in an attempt to block angiogenesis.
blood to transport oxygen. In healthy people, when
The drugs Torisel (Pfizer; New York, NY) and
the number of red blood cells in circulation drops,
Zortress (Novartis; Basel, Switzerland) stop mTOR, a
the kidney releases a hormone called erythropoietin,
protein that activates HIF. It is thought that the anti-
which stimulates the bone marrow to produce more red
angiogenesis effects of these mTOR inhibitors are a
blood cells. In chronic kidney disease (CKD), the kidneys
result of suppressing HIF. Torisel is FDA-approved for the
don’t produce adequate amounts of erythropoietin in
treatment of renal cell cancer; Zortress is FDA-approved

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for advanced kidney cancer, metastatic pancreatic reduced atmospheric pressure. Every breath taken at
neuroendocrine tumors, hormone-positive, and HER2- higher elevations delivers less oxygen than it would at
negative breast cancer. lower elevations, creating a slightly hypoxic environment
inside the athlete’s cells. This hypoxia increases the
COCKTAIL FODDER: THE levels of HIF, leading to more erythropoietin and
HIF ADVANTAGE subsequent red blood cell production. The enhanced
You may have heard of “altitude training” — the oxygen-carrying capacity lasts for about ten to twenty
practice of training at high altitudes in order to increase days after returning to lower elevations, so an athlete
performance, especially in endurance events such as who trains at a higher altitude and then competes at sea
long distance running or cycling. Altitude training works level will have an advantage over those who complete all
because at elevations higher than about 5,000 feet there of their training at sea level.
are fewer oxygen molecules per volume of air due to

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THE PARP RACE IS ON Life Science Training from Industry Experts

PARP1 INHIBITOR LINEUP the cells in which they would normally do their job are
prone to sustaining DNA damage at a much higher rate
PARP1 inhibitors are making a strong statement!
than normal. This higher rate of DNA damage increases
Tesaro’s (Waltham, MA) just-approved Zejula
the chances of cancer developing in those cells. BRCA1/
has garnered predictions of blockbuster status.
BRCA2 positive cancer is cancer that is associated with
AstraZeneca’s (Cambridge, UK) Lynparza was the first
mutations in the BRCA1/BRCA2 genes. The mutations are
PARP1 inhibitor to make it to market back in 2014, and
most strongly associated with breast and ovarian cancer,
their recent clinical trial results showed significant
but are also associated with increased risk of developing
survival benefit in ovarian cancer. Clovis Oncology
stomach, pancreatic, prostate, melanoma, leukemia,
(Boulder, CO) achieved the second FDA approval of a
lymphoma, and colon cancer.
PARP1 inhibitor with Rubraca in December 2016. AbbVie
(North Chicago, IL) and Medivation (San Francisco, CA) THE POINT OF PARP
both have PARP1 inhibitors in late-stage development.
Poly ADP ribose polymerase 1 (PARP1) is a DNA repair
The race is in full swing, so let’s pick up the science of protein. By stopping the PARP1 repair pathway in cells
PARP1 inhibition. already deficient in BRCA1/BRCA2-mediated repair,
cancer cells become extremely vulnerable to DNA
DNA DAMAGE RUNS DEEP damage. Because of this, DNA damage accumulates
Simply put, PARP1 inhibitors work by exploiting the and triggers apoptosis. A PARP1 inhibitor is usually
cellular pathways found in DNA damage repair. So, how administered in combination with chemotherapy or
exactly does DNA get damaged? radiation therapy, which increases the incidence of
DNA incurs approximately 10,000 to 1,000,000 apoptosis-triggering DNA damage. Healthy cells, which
“molecular lesions” per day from breaks or “nicks” to the still have BRCA repair pathways intact, are less sensitive
double helix, or chemical modification to the A, C, G, or to additional DNA damage.
T bases. This may sound high — but remember, our DNA
contains 6 billion bases (3 billion base pairs), so this is
equivalent to .001% to .1% of the total DNA in each cell.
This damage occurs as a result of normal DNA replication
errors and environmental exposures, such as ultraviolet
radiation, X-rays, and chemicals.
The good news is our cells have mechanisms to fight
against this damage before it causes harm. DNA repair
proteins find and fix different types of DNA damage. If
DNA damage exceeds a threshold amount (beyond which
repair is possible) a protein called p53 triggers cell death
— also known as apoptosis. DNA repair proteins prevent
What cancers are PARP1 inhibitors aiming to fight?
errant cells from turning into cancerous cells, a likely
AstraZeneca’s Lynparza and Clovis Oncology’s Rubraca
outcome if the damage accumulates in genes important
are both approved for ovarian cancer. Zejula (Tesaro)
for regulating cell growth and division.
targets ovarian, fallopian tube, and primary peritoneal
cancer. The Phase III clinical pipeline includes:
BEHIND BRCA
• Lynparza: Prostate, gastric, breast, and pancreatic
Arguably the most famous DNA repair proteins, BRCA1
• Rubraca: Prostate
and BRCA2, are found in breast and ovarian cells. If
• Talazoparib (Medivation): Breast
these repair proteins themselves are non-functional,
• Veliparib (AbbVie): Breast, lung, and ovarian

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 • Zejula: Breast A DNA mutation is a change to the actual base sequence
(A, C, G, or T). Mutations can arise if DNA damage
BEYOND CANCER is not corrected. Recall that in undamaged DNA, an
Preclinical research suggests that PARP1 inhibitors “A” base always pairs with a “T” base, and a “C” base
may also be relevant to other disease areas, such as always pairs with a “G” base. These base-pairing rules
autoimmune and inflammatory disorders. PARP1 has are what enable DNA to replicate faithfully from one
been shown to play a role in activating proteins that generation of cells to the next. However, uncorrected
drive inflammation. Preclinical models demonstrate DNA damage may cause that “A” base to mistakenly pair
that in cases without the PARP1 gene, subjects were with a “G” during replication; or a “C” to pair with a “T.”
less vulnerable to rheumatoid arthritis than with the This results in a sequence change – a mutation - in the
gene. Inhibiting PARP1 resulted in reduced signs of replicated DNA. The gene now provides incorrect genetic
inflammation in models of multiple sclerosis, irritable information to the cell.
bowel disease, and allergic airway inflammation.

EASILY CONFUSED: DNA

DAMAGE VS. DNA MUTATION
BRCA1, BRCA2, PARP1, and other DNA-repair proteins
correct DNA damage, but they don’t fix mutations.
What’s the difference?
DNA damage refers to alterations in the chemical
structure of DNA. This may mean a break in the
DNA strand, a substitution to one of the bases that
make up DNA (A, C, G, or T), or even a missing base.
These changes are detected and corrected by DNA
repair enzymes.

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PILLS, PROTEINS & PEPTIDES Life Science Training from Industry Experts

PROMISING PEPTIDE THERAPIES which include sensitivity to digestive enzymes, delivery

by injection, and high specificity for their target.
The front runners in the game of drug delivery include
small molecule and large molecule drugs, but there is
another class that lands right in between: peptides.
Several companies, including Rhythm Pharmaceuticals
(Boston, MA), Kalos Therapeutics (San Diego, CA),
Aileron Therapeutics (Cambridge, MA), and Bicycle
Therapeutics (Cambridge, MA) have emerged as
prominent players in the peptide arena.
Let’s review the differences between the drug classes
and explain where peptides fit into the picture. Then
we’ll take a spectator’s interest in the companies and
products leading the charge in peptides therapeutics.
Examples of peptide drugs on the market today include
EASILY CONFUSED: SMALL MOLECULE glucagon-like peptide-1 (GLP-1) receptor activators, such
VS. LARGE MOLECULE VS. PEPTIDE as Byetta (AstraZeneca; Cambridge, England), Victoza
(Novo Nordisk; Bagsvaerd, Denmark), and Trulicity
Small molecule drugs are chemically synthesized — made
(Eli Lilly; Indianapolis, Indiana). These peptide drugs
by a series of chemical reactions in the lab. They are
work by interacting with a receptor on the surface of
typically taken as a pill or capsule. The pill or capsule
pancreatic beta cells and stimulate the release of insulin
dissolves in the gastrointestinal tract and the active
for diabetes.
ingredient is easily absorbed into the bloodstream
via the intestinal wall. The molecules that make up
IN THE RHYTHM
these drugs are so tiny they are able to penetrate cell
membranes and get inside of cells. Rhythm Pharmaceuticals is prepping to enter Phase
III clinical studies of their anti-obesity peptide drug
In contrast, large molecule drugs — protein-based
setmalnotide. Designated as a breakthrough therapy
therapeutics known as biologics — are made by living
by the FDA, early clinical trial results in rare genetic
cells. They must be administered via injection because
forms of obesity were promising, helping to attract $41
they will be destroyed by digestive enzymes in the
million from key investors including Pfizer Venture
gastrointestinal tract if given orally. Their large size,
Investments and Third Rock Ventures to fund the
anywhere from 50 to 1,000 times larger than a typical
upcoming Phase III.
small molecule drug, makes it impossible for them to
penetrate cells. On the flip side, large molecules are Setmalnotide works by activating the melanocortin-4
highly specific for their target — typically a cell-surface receptor (MC4R), a receptor present on the surface of
receptor on the outside of the cell. cells in the hypothalamus of the brain, a region involved
in regulating both appetite and satiety. Mutations in
The FDA defines a peptide therapeutic as a chain of
MC4R that result in reduced activation are the most
amino acids (the building blocks of proteins) containing
common genetic cause of obesity, and account for
40 amino acids or less, and regulates them as small
approximately 6-8% of obesity cases.
molecules. Peptide therapeutics are similar to small
molecule drugs in that they can be synthesized in the
KALOS FIGHTS CANCER
lab using a peptide synthesis machine — a machine that
links amino acids together in a specified order. Peptides Kalos Therapeutics has a peptide drug in development
also share key characteristics with large molecule drugs based on a straightforward observation: despite

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constant activity, heart muscles don’t get bigger, and cancer cells but is inactivated in a range of malignancies.
cancers of the heart are extremely rare. At least part of ALRN-6924 is in Phase II clinical studies for lymphoma.
the reason for this is a peptide known as atrial natriuretic The company is pursuing the development of stapled
peptide (ANP), which is produced in the heart. It helps peptides in a range of therapeutic areas, including
to control cell growth and division, making sure that inflammation and endocrine and metabolic diseases.
the heart doesn’t get too big for the chest. Since cancer Bicycle Therapeutics also uses chemically linked
is caused by out-of-control cell growth and division, a peptides to increase stability, target interaction, and
connection was made: perhaps these peptides could play penetrate cells. Their peptides are formed — using a
a role in controlling tumor cell growth. chemical linker — into the shape of a bicycle.
Kalos Therapeutics has identified a portion of ANP and Bicycle’s lead candidate, BT1718, is a “bicycle drug
is synthesizing and testing it as a potential anti-cancer conjugate” — a bicyclic peptide with a toxic drug
agent. Dubbed KTH-22, the agent is cytostatic, meaning attached. The peptide targets a protein called
it halts the growth and division of cancer cells, but “membrane type 1 matrix metalloproteinase” (MT1-
does not directly kill them as a cytotoxic (toxic to cells) MMP) which is overexpressed in many tumors. BT1718
agent would. KTH-22 is in preclinical research, with data delivers its toxic payload to tumors overexpressing MT1-
supporting its use in the treatment of pancreatic and MMP. Preclinical trials have shown high efficacy against
ovarian cancers. these tumors, and clinical trials are expected to start by
the end of 2017.
STAPLES & BICYCLES
Most peptide therapeutics do not penetrate cell
membranes. Designing peptides that could enter cells
would truly endow them with the best characteristics
of both large and small molecule therapies. Aileron
Therapeutics and Bicycle Therapeutics are aiming to
do just that.
Aileron Therapeutics is developing “stapled peptides.”
These peptides are synthesized according to an
optimized amino acid sequence. Next, a chemical linker
is used to connect two amino acids within the chain,
creating a folded or “stapled” version of the peptide.
These stapled peptides still recognize their target Already capable of affecting a range of therapeutic
protein, are more stable, and better able to penetrate targets with high specificity, continued innovations in
cell membranes than the linear versions. peptide design and delivery should make this class of
drug an important player.
Aileron’s leading stapled peptide candidate, ALRN-
6924, activates p53, a protein that triggers cell death in

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PUTTING THE NA IN DNA Life Science Training from Industry Experts

NUCLEIC ACID THERAPEUTICS complementary base pairs — “A” complements “T”

and “C” complements “G.”
Small molecule, peptide, and biologic drugs aren’t the
only players in the game of drug development. A fourth • Bases include Adenine (A), Cytosine (C), Guanine (G),
class of therapeutics differs from all three of these: Thymidine (T).
nucleic acid-based drugs. These drugs are rising in RNA
prominence due to their potential to specifically target a • The ribose sugar group is a less chemically stable
wide range of diseases, including various types of cancer, sugar group because “ribose” has a highly reactive
autoimmune, and infectious diseases. Companies like oxygen atom.
Moderna (Cambridge, MA) are garnering unprecedented
• RNA is typically single stranded.
investor interest, while improvements in delivery
methods have increased the efficacy of nucleic acid- • Bases include Adenine (A), Cytosine (C), Guanine (G),
based therapies. Uracil (U).
In this WEEKLY — the first of a two part series — we’ll
dig up the different types of nucleic acids and unearth
mRNA-based therapeutics in development.

TERM OF THE WEEK: NUCLEIC ACID

Nucleic acids are long chains of repeating units of
nucleotides. Nucleotides are made up of a phosphate
group, a sugar group, and a base.

WHY MRNA?
You probably recognize DNA as the molecule of heredity,
and may recall that it provides cells with the instructions
for making proteins. Enter messenger RNA (mRNA) — the
literal messenger that relays the DNA instruction to the
ribosome where the protein-making process takes place.

There are two types of nucleic acids: DNA and RNA.

The nucleotides (or building blocks) of these two
varieties of nucleic acids are quite similar, but there are
marked differences.
DNA
• The deoxyribose is a more chemically stable sugar
group because “deoxyribose” lacks the highly
reactive oxygen atom.
• Two individual strands of linked nucleotides join
together to make the double helix by forming

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So, why all this talk about mRNA? Well, protein IN THE PIPELINE
therapeutics — the injectable protein-based drugs
Moderna Therapeutics (Cambridge, MA) has
discussed last week — have revolutionized the
received nearly $2 billion to fund ongoing mRNA drug
treatment of a range of diseases, from diabetes to
development. The company now has five different
cancer to autoimmune disorders. However, they are
products in Phase I clinical studies. Four of these are
time consuming and expensive to produce. Cells must be
vaccine candidates: two against different strains of
engineered to develop the desired protein, then grown in
influenza virus, one against Zika virus, and one against
large (thousands of liters) tanks. Finally, the therapeutic
an undisclosed target in partnership with Merck
protein must be painstakingly purified away from other
(Kenilworth, New Jersey). A mRNA-based vaccine uses
proteins and cellular debris in the cell.
lipid nanoparticles to deliver the instructions for making
What if we could eliminate the huge biomanufacturing a particular viral protein to a cell. The cell then makes the
tanks and just have the patient make the therapeutic viral protein and displays segments of it on its surface,
protein using their own cells? That is the idea behind activating an immune response to fight infection.
mRNA therapeutics — figure out a way to provide the
The fifth drug for which Moderna has initiated clinical
information contained in mRNA directly to the patient’s
trials — in partnership with AstraZeneca (Cambridge,
ribosomes and let the patient’s cells do the work. Not
UK) — is a mRNA that codes for the protein known as
only would this be more efficient, it would also enable
vascular endothelial growth factor, or VEGF. This protein
therapeutic proteins to be introduced directly inside cells
promotes the growth of blood vessels, and may help
or embed into the cell membrane. Recall that protein
to improve blood supply in cardiac tissue after a heart
therapeutics injected into the bloodstream are too large
attack, or in diabetic wound healing.
to enter cells and are limited to interacting with proteins
on the surface of cells or in the blood. CureVac (Tubingen, Germany) is focused on mRNA
vaccines as well, with a prostate cancer therapeutic
THEORY VS. REALITY vaccine in Phase II clinical studies. Therapeutic vaccines
Like much in biotech, the concept of mRNA-based train the patient’s immune system to recognize a specific
therapeutics is elegant in theory, yet rough in reality. cancer associated protein, priming immune cells to
attack the tumor that bears those proteins. CureVac
• Reason 1: The relative instability of the mRNA
also has a mRNA-based rabies vaccine in Phase I clinical
molecule itself; mRNA traveling through the
studies, with several more infectious disease and
bloodstream would typically be degraded by
therapeutic cancer vaccines in preclinical development.
nucleases — enzymes that break down nucleic acids.
Other companies to watch in this space include:
• Reason 2: “Foreign mRNA” coming from outside of the
cell could trigger an immune response; our immune • BioNTech (Mainz, Germany): Phase I studies
systems have evolved to recognize foreign mRNA completed on a mRNA-based therapeutic vaccine
as bad. for melanoma; preparing to enter clinical studies
on therapeutic cancer vaccines for head and neck
• Reason 3: Delivery of mRNA therapy is difficult.
cancer and personalized vaccines.
Right now the approach that appears to be having
the most success is encasing the mRNA in a lipid • Arcturus (San Diego, CA): Preclinical development of
nanoparticle for delivery to cells. mRNA drugs to treat protein deficiency disorders.

Bringing mRNA drugs to market involves designing • RaNA (Cambridge, MA): Preclinical development of
chemically modified mRNA that is more stable (resistant mRNA drugs to treat protein deficiency disorders.
to nucleases) and less visible to immune cells than mRNA drugs show much promise and we will continue
unmodified mRNA. These modified mRNA molecules are to closely follow this area for new developments. Next
called “nucleotide analogs” because they are similar but week, we’ll continue our discussion of nucleic acid-based
different from naturally occurring nucleotides. therapeutics as we look at additional types of RNA and
DNA based drugs.

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RNA THERAPEUTICS MARCH ONWARD Life Science Training from Industry Experts

TAKING STEPS WITH ANTISENSE renewed itself in 2013 thanks to the FDA approval of
Ionis Pharmaceuticals’ (Carlsbad, CA) Kynamro for the
With their high specificity and relative low manufacturing
treatment of familial hypercholesterolemia.
cost, RNA therapeutics may be tomorrow’s biotech
sweetheart. In fact, chances are good that previously Kynamro targets apolipoprotein B, a key component of
“undruggable” targets that cannot be accessed by small LDL cholesterol, to lower cholesterol levels:
or large molecule drugs, are now within reach. However,
the main roadblock continues to be delivery—getting
the RNA drug where it needs to be, in high enough
concentrations, to be effective.
Last week we looked at the emerging class of
nucleic acid-based drugs known as messenger RNA
therapeutics. This week, we’ll continue our discussion
by examining the RNA therapeutic known as antisense.
Let’s march!

RNA RELAY
Messenger RNA (mRNA) is a molecule that carries a
copy of the protein-making instructions from DNA in the MEET INTRONS & EXONS
nucleus of a cell to the ribosomes located just outside Exondys 51 (Sarepta Therapeutics; Cambridge, MA)
of the nucleus. The information contained in mRNA is and Spinraza (Ionis and Biogen; Cambridge, MA) are
used by the ribosomes to assemble a protein. Without recently-approved antisense drugs that work by altering
mRNA relaying the instructions, there would be no how the cell processes pre-mRNA — an immature
protein produced. template of mRNA. Pre-mRNA is a long strand of RNA
from which specific pieces are cut out or added in before
ANTISENSE EXPLAINED becoming the mature mRNA that is able to relay the
Antisense drugs are short (between 10 and 30 protein-making instructions from DNA to the ribosomes.
nucleotides), synthetic pieces of nucleic acid whose Think of pre-RNA as a master grocery list that includes
sequence is complementary to the mRNA that codes for all of the most common foods you buy over the year.
a disease-associated protein. When the antisense drug Envision mRNA as your adjusted grocery list that
enters a patient’s cells, it binds to the disease-causing includes only what you need to buy for this week.
mRNA. This binding triggers an enzyme called RNAse H
When pre-mRNA is first produced, it contains two types
to destroy the double-stranded antisense-mRNA hybrid.
of sections that are either cut out or left in depending on
Our bodies recognize that double-stranded RNA as a
the instructions:
mistake and destroys it. Without the mRNA, the disease-
associated protein simply is not made — stopping • Introns: regions cut out of the final mRNA; grocery
disease in its tracks. items you delete from your master list because you
don’t need them this week.
The history of antisense in drug development is quite
fickle. Over the years the inability to get antisense • Exons: regions included in the final mRNA; grocery
drugs to accumulate at therapeutically effective items you keep from your master list because you
concentrations in the target tissue was the undoing of need them this week.
many clinical trials. But the love affair with antisense

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 SMA patients don’t produce enough of a protein called
spinal motor neuron (SMN) because it is missing an exon;
the right item was left off during the list-making process.
Spinraza fixes this error, in other words, the antisense
drug glues itself to the exon which needs to stay on so it
can remain on the list. This results in the production of a
full-length mRNA and a more functional SMN protein.

Duchenne’s muscular dystrophy (DMD) and spinal

muscular atrophy (SMA) are diseases in which errors
occur in the processing of pre-mRNA, resulting in
proteins that are not fully functional. Harking back to our
grocery list analogy, this means your weekly grocery list
is incorrect. Some wrong items were left on and some right
were left off, resulting in a failed shopping trip.
Antisense drugs alter the way pre-RNA is processed
by either including or excluding exons to make a more
functional protein. Antisense technology fixes the weekly THE FUTURE OF ANTISENSE
grocery list. The future of antisense looks bright. By targeting the
In DMD, the wrong items were left on during the list- right tissues (the most promising targets are in the liver)
making process. Exondys 51 is an antisense drug that and developing more stable formulations, we can expect
binds to the “wrong item” to ensure it stays off. In to see more success stories. Check out some of the
reality, it cuts a mutated exon out of the dystrophin antisense drugs currently in clinical development:
gene. By cutting out this exon, DMD patients produce
a more functional dystrophin protein than the disease-
associated version. The dystrophin protein maintains
muscle integrity.

The complex yet fascinating world of antisense marches

forward. They promise several key advantages, including
high specificity, low manufacturing costs, and the
potential to target previously “undruggable” targets. We
here at the WEEKLY look forward to reporting on new
approvals in this arena.

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DECODING YOUR GENES Life Science Training from Industry Experts

THE SKINNY ON DNA TESTING Many SNPs have no significant impact on an individual’s
health, but other SNPs are associated with disease
23andMe (Mountain View, CA) recently found itself back
susceptibility; having one variant instead of another
in the limelight after the disease risk section of its mail-in
makes someone more likely to get a particular disease.
DNA kit received an OK from the FDA.
SNP genotyping is characterizing the SNP profile of
The Silicon Valley biotech had to halt sales of its
an individual — finding out which A, C, G, or Ts are
direct-to-consumer genetics testing back in 2013 after
in positions differing from the norm. In the case of
regulatory officials grew concerned over marketing
23andMe, that information is used to assess disease
claims and the possibility of consumers misinterpreting
risk, carrier status for certain genetic diseases, wellness
the test results. 23andMe rebooted a limited part of their
information, and ancestry.
genetics section in 2015 after a preliminary go ahead,
and last month marked a full return with the approval of A CHIP OFF THE OLD GENE
their Genetic Health Risk (GHR) reports.
23andMe’s core technology involves the use of DNA
In this issue, we’ll get the skinny on the science microarrays, also known as DNA “gene chips.” The
driving 23andMe. technology relies on the very specific base pairing rules
followed by double-stranded DNA: A’s always pair with
TERM OF THE WEEK: SNP GENOTYPING T’s, and C’s always pair with G’s.
23andMe’s moniker is a nod to the 23 pairs of
A DNA microarray is simply a tiny piece of glass or silica
chromosomes that make up the human genome. Recall
that has had a microscopic checkerboard etched into it
the human genome is all the DNA found in a human
— each square is about 11 micrometers (10-6 meters) by
cell. The tests, which are available to anyone with a
11 micrometers big, just big enough to hold one single-
mailbox and a credit card, rely on a technique called
stranded piece of DNA.
SNP genotyping.
“SNP” (pronounced snip) stands for “single nucleotide
polymorphism” — a change in a single DNA nucleotide
(A, C, G, or T) that occurs at a specific position in the
genome. An example of a SNP is demonstrated in the
image below: within a specific gene sequence the most
common variant for individuals is a “T” at a particular
location, but in some individuals the “T” has been
replaced by a “C” SNP.

Each square represents one gene, or gene variant, within

the human genome. The entire gene sequence need not
be present — typically, between 25 and 60 bases are
used (though we are only showing 6 in our image above).
Computer software keeps track of the location of gene
sequences within the array. In this manner, the specific
variants within each DNA sample can be identified.

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EASILY CONFUSED: GENOME Mutations in susceptibility genes, in contrast, do not
SEQUENCING VS. SNP GENOTYPING necessarily mean the patient has or will develop the
disease, it only means the patient is at risk for developing
The term DNA sequencing is often mistakenly used when
the disease. One well-characterized susceptibility gene
the correct term is SNP genotyping.
is apolipoprotein E (ApoE). The ApoE4 variant of the
• DNA sequencing means to determine the order ApoE gene is associated with a higher risk of Alzheimer’s
of every single base pair in a given gene (gene disease (AD). Those with two copies of ApoE4 may have
sequencing) or in an entire genome (whole as much as 20 times the risk of developing AD; however,
genome sequencing). some individuals with two copies never develop the
• SNP genotyping means to identify single base disease according to statistical studies of different ApoE
changes between a given gene sequence and a variants. On the other hand, another variant — ApoE2 —
reference sequence. may reduce the risk of developing AD.
By identifying which ApoE gene variant an individual
MUTATIONS & SUSCEPTIBILITY GENES has, researchers can say whether or not that person has
To qualify as a diagnostic test in the eyes of the FDA, an average risk, a higher than average risk, or a lower
genetic testing companies must show that a specific DNA than average risk of developing AD — but they cannot
sequence can confirm whether one has (or doesn’t have) definitively diagnose or predict AD onset.
a particular disease.
For example, Huntington’s disease is caused by a known
mutation in the huntingtin gene. If a specific mutation
is detected, a doctor can say with certainty whether the
patient has or will develop Huntington’s disease. It is a
dominant genetic disorder, meaning that one copy of the
disease-associated gene variant is enough to cause the
disease (recall that we have two copies of each gene, one
from each parent).
Most genetic disorders are recessive — meaning two
copies (one from each parent) of the disease-associated
gene must be present to actually develop the disease. 23andMe now offers 10 Genetic Health Risk tests that
If someone has only one copy of the gene variant that look at susceptibility genes, including ApoE4. Part of the
causes cystic fibrosis, they are deemed a “carrier” — they FDA’s earlier concerns about consumer misinterpretation
themselves do not have cystic fibrosis, but they have a involved the distinction between susceptibility and
25% chance of passing the disease on to their child if the diagnosis — making sure consumers understood the
other parent is a cystic fibrosis carrier. test results did not, for example, diagnose them with AD,
but merely reported their susceptibility in the form of a
statistical risk factor.
23andMe also offers carrier status tests for more than
40 different conditions. They also provide information on
gene variants associated with “wellness” — such things
as sensitivity to caffeine and lactose intolerance, as well
as non-health-related traits such as dry or wet earwax
and skin pigmentation.

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MORE THAN A SNP (in partnership with Genentech; South San Francisco,
CA), wherein they recruited patients with those diseases
Direct-to-consumer genetic testing is only a part of
to participate in identify SNPs more common in those
23andMe’s business. Since its founding in 2006, the
populations. In 2015, the company announced it
company has been collecting genetic data on patients
would begin to do its own drug discovery research and
along with their self-reports of symptoms and health
launched 23andMe Therapeutics.
status — about 85% of users from the 23andMe
database have opted in. From new options for consumer genomics to
breakthroughs in drug discovery, we expect 23andMe to
The company has also initiated research efforts around
stay in the news for some time to come.
specific diseases such as inflammatory bowel disease (in
partnership with Pfizer; New York, NY) and Parkinson’s

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PHAGE — MORE THAN JUST A PHASE Life Science Training from Industry Experts

OLD WAY OF FIGHTING other, resistance is much less likely to evolve as has been
BACTERIA RENEWED the case for antibiotics. Likewise, humans have safely
coexisted with bacteriophage for a long time, suggesting
One of the greatest public health challenges of the 21st
that there should be few safety issues with their use
century is antibiotic resistance, which occurs when a
as therapeutics.
few bacteria in a given population develop a genetic
mutation that enables them to survive — even in the Scientists have known about the bacteria-devouring
presence of antibiotics. ability of phage for about 100 years, but with the advent
of antibiotics in the late 1920s, medicine’s focus shifted
How do bacteria become drug resistant? Suppose a
to these new wonder drugs because they were easier to
particular antibiotic inhibits an enzyme required for
manufacture and test in controlled settings. Now that
bacterial replication. If one bacterium mutates so the
antibiotic resistance is emerging, so too is a renewed
enzyme has a slightly different shape, the antibiotic is
interest in bacteriophage, which are now starting to
no longer effective. The mutated bacterium lives on and
be manufactured and tested in a standardized way for
continues to replicate, even as all the others die off. Over
the first time. Let’s take a look at some of the biotech
time, this resistant strain becomes dominant, spreading
companies delving into the world of bacteriophage-
from person to person, remaining unchecked and
based therapeutics.
thriving. It is not uncommon for a strain of bacteria to
become resistant to several different antibiotics, giving THE COCKTAIL APPROACH
rise to the term multi-drug resistant bacteria.
The first multicenter clinical trial examining the use
In this issue of the WEEKLY, we’ll take a look at an of bacteriophage as antibacterial treatments was
entirely novel approach to fighting bacterial infections — initiated in 2015 by French biotech Pherecydes (Paris,
bacteriophage. France). Preparative work for the trial began in 2013 as
researchers established protocols for producing phage
TERM OF THE WEEK: BACTERIOPHAGE that met good manufacturing practice guidelines. The
A bacteriophage — also referred to as a phage — is a researchers are studying two different “cocktails” of
virus that infects bacteria. By attaching to a bacterium’s bacteriophage — mixtures of different bacteriophage
surface, a phage punches holes in the membrane and that have shown activity against different substrains
injects its own genetic material inside. The phage then of a particular bacteria in the lab. The first contains 13
replicates inside of the bacterium, creating so many new different phages targeting P. aeruginosa; the second, 12
viruses that the bacterium breaks open, releasing newly phages that target E. coli. Both are being evaluated for
produced viruses, which can then go on to infect other the treatment of burn wound-associated infections.
bacteria, continuing the cycle.
Other companies testing phage cocktails in human
The word “bacteriophage” is derived from the Greek patients include:
word phagein — “to devour.” So we can think of
• AmpliPhi Biosciences (Richmond, VA): Phage
bacteriophage as, literally, devouring bacteria — a
cocktail AB-SA01 is in Phase I clinical testing for
potentially very useful trait! Typically each phage is
antibiotic resistant S. aureus in two different clinical
specific for a type of bacteria, meaning that if adapted
settings: chronic rhinosinusitis as well as acute and
for therapeutic use, researchers can select viruses
chronic wound and skin infections.
that will only attack harmful bacteria, and leave the
many strains of “friendly” bacteria that make up our • TechnoPhage (Lisbon, Portugal): Phage cocktail TP-
gut microbiome alone. And since each type of phage 102 targets bacteria associated with chronic ulcers,
has coevolved for millennia with its chosen strain of respiratory and skin infections.
bacteria, each adapting and changing in response to the

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 • Intralytix (Baltimore, MD): Completed Phase I preclinical development include Synthetic Genomics
studies using a bacteriophage for the treatment of (San Diego, CA) and EnBiotix (Cambridge, MA).
infected wounds.
• EpiBiome (San Francisco, CA): Phage cocktails
THE COMPONENT APPROACH
targeting E. coli and S. dysenteriae diarrheal infections A third approach to tapping into the therapeutic power
in preclinical development. of bacteriophage lies in isolating the component or
components that make them toxic to bacteria. For
THE ENGINEERED APPROACH example, in order to inject their genome into bacteria,
Creating a phage cocktail can be a challenging phage must essentially punch a hole in the membrane,
biomanufacturing problem. An alternative approach to which is itself very damaging to the bacteria. The viral
combining beneficial characteristics of different viruses protein that creates these tears in the membrane
is to genetically engineer a synthetic virus that combines are called “lysins” — enzymes that essentially chew
the properties of multiple phages into a single genome. holes in the bacterial cell wall. ContraFect (Yonkers,
NY) has completed Phase I clinical studies of its drug
For example, scientists could insert genes into a phage
CF-301 — a lysin — for the treatment of S. aureus
genome to increase the range of bacteria subtypes
bloodstream infections.
an individual phage can attack, yet still maintain the
specificity that prevents the phage from raiding friendly Although still in the early days of clinical testing,
bacteria. Researchers could also add in genes to make bacteriophage offer the possibility of being a safe and
the bacteriophage’s antibacterial response even effective solution to the current antibiotic resistance
stronger. Companies with engineered bacteriophage in public health crisis.

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VIRUSES BLASTING CANCER Life Science Training from Industry Experts

ENGINEERING VIRUSES TO ATTACK against cancer. Let’s take a look at the unique features
of a selection of oncolytic viruses on the market and
Getting bacteria-eating viruses to combat antibiotic
in development.
resistance isn’t the only way viruses are being hacked
to defend team homo sapien. This week, we’ll turn our INSIDE OF IMLYGIC
attention to another benevolent use of viruses: cancer-
Viruses can be thought of as very simple packages of
fighters known as oncolytic viruses.
genetic material — DNA or RNA — encapsulated in a
ONCOLYTIC VIRUS PRIMER protein package. Like the human genome, viral genomes
code for proteins required by the virus. Some of these
Oncolytic viruses are an immunotherapy — a type of
proteins enable the virus to make copies of itself
therapy that harnesses the power of a patient’s immune
(replicate), or to evade the human immune response. It
system to combat a disease. Getting a virus to trigger
is often necessary to modify the viral genome in order to
the immune response to fight cancer is no easy task,
safely use a virus as a therapeutic, but how?
the process involves engineering the virus to selectively
infect and kill cancer cells. Oncolytic viruses are created Amgen’s (Thousand Oaks, CA) Imlygic is the only FDA
in the lab by genetically modifying existing viruses in at approved oncolytic virus, aiming to attack melanoma.
least two ways: The virus used in Imlygic is a modified herpes simplex 1
virus. The modifications made to Imlygic to ensure safety
• Making the virus safe by removing genes that cause
and efficacy include:
the virus to make people sick
• Deletion of viral gene ICP34.5. This gene codes for a
• Engineering viral surface proteins so the virus
protein that enables the virus to replicate in human
recognizes and binds to the cell receptors of
cells by blocking a human protein known as PKR.
cancerous cells, disregarding the healthy, non-
PKR prevents viral replication. It is less active in
cancerous cells
most tumor cells, so this makes the virus able to
The oncolytic virus follows the same life cycle as any selectively replicate in tumor cells.
virus—once inside the human body it hunts down,
• Deletion of viral gene ICP47. This gene codes for a
attaches to, and enters its host cell. In this case, the host
viral protein that thwarts the immune response by
happens to be cancer cells! The virally infected cancer
turning off a process called antigen presentation.
cells are destroyed via the process of cell lysis—as the
Normally, one of the key ways the immune system
oncolytic virus multiplies inside of the cells, it causes
“knows” to attack a virally-infected cell is by
the cancer cells to burst open which kills them. Spewing
recognizing antigens (or fragments of viral proteins)
from the burst cells are new infectious viruses that
displayed on the infected cell’s surface. Turning
further target remaining tumor cells. The presence of a
this process off helps the virus evade the immune
replicating virus also activates the immune response, so
system. Turning it back on prompts the immune
the cancerous area is further attacked.
system to attack virus-infected tumor cells.
Additional modifications may also be made to the virus,
• Activating the earlier expression of the viral gene
depending on the characteristics of the targeted cancer.
US11, resulting in increased viral replication in
For example, an oncolytic virus might be modified to
tumor cells.
produce proteins that stimulate the immune system or
directly attack the tumor. • Insertion of a gene for the human protein GM-
CSF, which activates the immune system, aiding in
Most oncolytic viruses are tested both as “stand-
the overall immune response toward the tumor
alone treatments” and “in-combination with other
triggered by viral infection.
immunotherapies” — such as checkpoint inhibitor
therapies — to help fully activate the immune response

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Taken together, these modifications create a virus that for “therapeutic payloads” — proteins that will boost
selectively replicates in tumor cells, resulting in their the patient’s immune response to the cancer, or even
direct destruction as well as activation of a host immune therapeutic antibodies that will then be produced inside
response targeting the virus-infected tumor cells. of the cancer cell. This approach is a clever response
to the fact that due to their relatively large size, most
ONCORUS ON THE OFFENSE therapeutic antibodies are not able to completely
Oncorus (Cambridge, MA) is also developing a modified penetrate solid tumors. Using an oncolytic virus to
herpes virus, ONCR-001, for the treatment of cancers, penetrate the tumor and deliver genes instructing the
including the notoriously difficult-to-treat brain cancer, tumor itself to make the antibody could be a game-
glioblastoma. Like Imlygic, ONCR-001 has been modified changing work around.
to selectively target tumor cells. Unlike Imlygic, ONCR- Finally, Genelux is also creating engineered virions that
001 retains all viral genes needed for viral replication. incorporate “imaging” proteins. For example, GL-ONC1
So, how is safety maintained in the presence of an delivers a gene for a fluorescent protein directly to tumor
oncolytic virus that is actively replicating? Oncorus cells. As the virus replicates inside of the tumor, the
scientists have figured out a clever way to take fluorescent signal increases. In preclinical animal testing,
advantage of a key difference: the types of microRNAs this has allowed non-invasive detection and imaging of
produced by healthy cells vs. cancer cells. tumor progression and regression in real time, and may
one day be a powerful guide to physicians monitoring
MicroRNAs are a type of “regulatory RNA” that promotes
cancer patients. GL-ONC1 has successfully completed
the degradation of a target messenger RNA (mRNA —
Phase I and is preparing to enter Phase II for a variety of
the RNA that gets turned into proteins). This means that
solid tumors including peritoneal carcinomatosis.
different target sequences will be recognized in healthy
cells vs. cancer cells. By engineering sequences that
ONCOLYTIC PIPELINE
microRNAs from healthy cells will recognize, Oncorus
scientists can ensure the viral mRNA will be destroyed With the approval of Imlygic in 2015, biotech companies,
in any healthy cells it infects. No viral mRNA, no viral investors, and regulatory officials have recognized the
proteins, no virus. Because these microRNAs are not strong potential of oncolytic viruses to treat cancer.
present in tumor cells, ONCR-001 is able to produce viral The race is on to get the next oncolytic virus through
proteins and new viral particles freely when it infects the clinic:
those target cells. ONCR-001 has shown strong ability in
fighting glioblastoma in preclinical models.

THE GENESIS OF GENELUX

San Diego-based Genelux is adapting the vaccinia virus
as an oncolytic virus for the treatment of a variety of
solid tumors. Vaccinia is the scientific name for the
cowpox virus — the virus that is used as a vaccine for
smallpox. Because of its decades-long use as a vaccine,
researchers have confidence the virus is safe to use
in humans, although the modified version must still
undergo safety testing.
We expect to see this new therapeutic class become
Their lead product, GL-ONC1, selectively replicates in increasingly common, opening up novel approaches for a
tumor cells and tumor-associated blood vessels, directly whole range of cancers.
killing tumors while cutting off their blood supply. The
company is also developing oncolytic viruses with genes

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NANOBODIES: THESE ARE NOT
YOUR MOTHER’S MABS Life Science Training from Industry Experts

THE DRUG KINGPINS the immune system to attack the tumor. Antibodies are
also selected for their ability to inhibit (stop the activity
Monoclonal antibodies (mAbs) are the undisputed drug
of) a particular protein. For example, the breast cancer
kingpins. In 2013, the mAb market raked in $75 billion in
drug Herceptin (Genentech, South San Francisco)
combined sales, covering a whole range of indications
inhibits the activity of the growth factor receptor HER2
from cancer and infectious disease, to autoimmune
by preventing it from interacting with the growth
disorders, and even high cholesterol.
factor HER2:
Despite the success, mAbs have one chink in their armor:
they cannot enter cells due to their large size, hampering
their range as therapeutics. To date, mAbs can only
target proteins on the surface of cells, such as receptor
proteins, or proteins circulating in the bloodstream, such
as inflammatory cytokines. The development of cell-
penetrating mAbs would open up a world of therapeutic
targets and patient benefits.
Let’s review the fundamentals of therapeutic antibodies
and explore a new type of therapeutic antibody that may
be able to go where no antibody has gone before.

MAB RECAP By developing antibodies that can enter cells, this

inhibitory power can be used against targets inside of the
Antibodies are proteins naturally produced by our
cell. Let’s take a look at the newest contender.
immune system to help defend against foreign invaders
such as viruses and bacteria. Each antibody produced
NANOBODIES
has a unique shape that enables it to recognize unique
targets (antigens) which are typically proteins on the Scientists at Ablynx (Ghent, Belgium) are developing
surface of pathogens. By binding to these pathogens, a new type of therapeutic antibody from an unlikely
antibodies act as a flag to alert the rest of the immune source — camels and llamas, members of the biological
system to attack. family Camelidae. These antibodies are structurally
and functionally very similar to human antibodies,
with a few important differences that could add up to
something big!
Like all antibodies, Camelidae antibodies work because
they have a specific shape that enables them to
recognize and bind to a specific target. However, they are
a tenth the size of other mammalian antibodies — giving
rise to the moniker “Nanobodies.” Nanobodies have the
ability to recognize targets hidden inside of cells. Their
small size may potentially also enable them to cross
the challenging blood brain barrier, or penetrate the
interior cells of tumors – two activities that conventional
Antibody therapeutics also rely on the ability of antibody therapies lack.
antibodies to interact with a specific target. Scientists
In addition to their small size, Nanobodies also exhibit
have developed antibodies that recognize and bind
a less complex structure overall. Because of this, they
proteins on the surface of tumor cells, thereby alerting

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have been successfully produced in bacterial cells. If that stimulates the immune response; inhibiting the
Nanobodies can be scaled-up, it would significantly immune system may prove a useful treatment for
reduce production costs as compared to standard autoimmune disorders. Vobarilizumab is being tested
antibody production in mammalian cells. Preliminary for the treatment of rheumatoid arthritis and lupus, in
studies in mice also suggest that Nanobodies can be partnership with AbbVie (North Chicago IL). Next up
maintained in the stomach and intestine — opening up is ALX-0171, which binds the fusion (F) protein on the
the possibility of oral delivery for some indications such surface of the respiratory syncytial virus (RSV). The F
as Crohn’s disease. protein enables RSV to lock onto lung cells. ALX-0171 is
expected to interfere with the interaction of F protein
IN THE CLINIC and lung cells, thereby preventing RSV infection.
We can expect to see the first Nanobodies on the In partnership with Boehringer Ingelheim (Ingelheim,
market this year. In February of 2017, Ablynx submitted Germany), Ablynx is also entering the oncology space
an application to the European Medicines Agency for with Phase I testing of a Nanobody that inhibits the
the first of its Nanobody therapeutics, caplacizumab. vascular endothelial growth factor (VEGF) protein. VEGF
Caplacizumab is being tested for the treatment is a growth factor secreted by tumor cells to encourage
for a rare disease known as acquired thrombotic the growth of blood vessels into the tumor, a process
thrombocytopenic purpura (aTTP), a blood-coagulation called angiogenesis. By inhibiting VEGF and angiogenesis,
disorder that results in extensive microscopic clots the flow of blood and nutrients into the tumor is
forming in small blood vessels throughout the body. stopped, essentially starving the tumor. It is hoped that
The disease is triggered by excess von Willebrand Nanobodies may be even better angiogenesis inhibitors
factor (vWF), a protein that initiates blood clotting. than monoclonal antibodies have proven to be, due to
Caplacizumab inhibits vWF, thereby preventing their enhanced tumor-penetrating abilities.
clot formation.
As Nanobodies continue to be tested for safety and
The company has two more Nanobody products in efficacy, a whole new kingdom of potential antibody
Phase II clinical testing: Vobarilizumab, which reduces targets may begin to emerge.
the activity of interleukin-6 (IL-6). IL-6 is a protein

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EPIGENOME: WRITING, READING & ERASING Life Science Training from Industry Experts

FOUNDATIONS OF EPIGENETICS A second type of modification is called acetylation — the

addition of an acetyl group (CH3 CO) to the histones.
Genetic mutations — changes in the order of the A, C, Acetylation loosens the association of the DNA with
G, and T nucleotide bases that make up a gene — have the histones, making the DNA more accessible to the
been the primary focus of cancer researchers over enzymes used in gene expression, ultimately increasing
the last several decades. By sussing out mutations protein production.
involved in regulating cell growth and division, scientists
better understand the molecular range of different
cancers and consequently develop more targeted and
effective therapeutics.
In recent years, another type of genetic variation has
captured the attention of researchers: epigenetic
modifications. Best characterized in cancer, epigenetic
changes are also thought to play a role in a range
of other diseases, including autoimmune disease,
cardiovascular disorders, diabetes, neurodegenerative
disorders such as Alzheimer’s disease, and potentially
even male infertility.
In this WEEKLY, we’ll tell the epigenetics story and discuss
Deacetylation — the removal of an acetyl group —
how it’s being used to develop new treatments.
increases the association or “grip” of the DNA around the
histones. Deacetylation makes the DNA less accessible
TERM OF THE WEEK: EPIGENETICS
to enzymes used in gene expression, thereby decreasing
Epigenetic modifications are changes to DNA that do the production of proteins.
not alter the actual gene sequence; they are chemical
modifications to the DNA itself. These changes typically ADDING IT ALL UP
affect gene expression, or how often the gene is read by the
cell. Epigenetic modification can occur either directly to the Epigenetic modification is a normal part of development.
nucleotide bases themselves (A, C, G, or T) or to the histones, This is in part why different genes are expressed in the
which are small proteins that package and order DNA. heart than, say, the liver — the two different tissue types
contain the same genome, but tissue-specific differences
in epigenetic modification lead to differences in gene
expression in the two tissues.
Problems may arise, however, if variations in epigenetic
modifications result in changes to gene expression.
If a cell or tissue type begins to make too much of a
protein that activates cell growth, for example, the cell
could begin to divide too often — potentially leading to
cancer. Alternatively, a cell could begin to make less of
a protective protein, for example, a “tumor suppressor”
protein (a protein that deactivates cell division), and
again cancer could ensue.
Epigenetic medicine seeks to identify disease-associated
One of the most common types of epigenetic
differences in epigenetic modifications, and to develop
modification is methylation — the addition of a methyl
drugs to restore the epigenome to that of healthy cells.
(CH3) group to cytosine (C) nucleotides. The end result:
methylation reduces or even blocks gene expression.

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BREAKING IT DOWN • Histone-methyltransferase (EZH2) inhibitors: EZH2’s
are also writers — these enzymes transfer methyl
Epigenetic drugs are small molecule drugs that target
groups to histone proteins. One EZH2 is associated
epigenetic regulators, or proteins that write, read, or
with over activity in a number of different cancers.
erase epigenetic modifications.
There are no EZH2 inhibitors currently approved, but
• Writers are the enzymes that make the chemical several are in development, including Constellation
modifications — methylation or acetylation
Pharmaceuticals’ (Cambridge, MA) CPI-1205
as described above — to DNA molecules or
in Phase I for advanced B-cell lymphomas, and
histone proteins.
Epizyme’s (Cambridge, MA) tazemat, currently
• Erasers are enzymes that remove these in Phase II for non-Hodgkin lymphoma, certain
chemical groups.
genetically-defined solid tumors, and mesothelioma.
• Readers are the proteins that detect and respond to
these modifications, causing the DNA to be more or THE NEW CLASS: READERS
less tightly wrapped around the histone protein. Any
A class of proteins called “Bromodomain and Extra
one of these proteins could be inhibited or activated
to affect changes in epigenetic modifications. Terminal motif” (BET) proteins are reader proteins. They
recognize and bind to specific patterns of acetylation on
OLD SCHOOL: WRITING & ERASING histone proteins. Upon binding, they recruit additional
proteins that regulate gene activity. Irregularities in
The disease that has been best classified in terms of
histone acetylation, then, may send the wrong message
epigenetic variations is cancer. Currently, there are five
to a BET protein. By inhibiting the interaction between
epigenetic drugs on the market to treat cancer, and more in
development. Those on the market fall into two categories: BET protein and histone proteins, researchers have
found that they can prevent incorrect messages from
• Histone deacetylase (HDAC) inhibitors: HDACs are
being received by the BET proteins. Currently, there
erasers — they remove acetyl groups from histone
are no BET inhibitors (BBI) approved, but several are in
proteins, resulting in increased expression of
clinical development. The farthest along is Resverlogix’s
associated genes. Three HDAC inhibitors have been
approved by the FDA: Zolinza (Merck; Kenilworth, (Calgary, Canada) apabetalone, which is in Phase III
NJ) and Istodax (Celgene; Summit, NJ) both treat testing for atherosclerosis and associated cardiovascular
cutaneous T-cell lymphoma, and Farydak (Novartis; disease. Additional BBIs in clinical development are
Basel, Switzerland) for the treatment of multiple shown in the table below:
myeloma. HDAC inhibitors in development include:

Epigenetics promises to change the way we look at the

• DNA-methyltransferase (DNMT) inhibitors: DNMT’s are human genome. Scientists have made great strides in
writers — they add methyl groups to DNA, resulting understanding how epigenetic modifications contribute
in decreased expression of associated genes. Two to both health and disease; however, a complete
DNMT inhibitors have been approved by the FDA: understanding of these modifications is still very much a
Vidaza (Celgene) and Dacogen (Otsuka; Tokyo, work in progress. As that work develops, researchers will
Japan). Both drugs are used to treat myelodysplastic undoubtedly uncover new drug targets and approaches
syndrome and acute myeloid leukemia. to disease management. Stay tuned!

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A SKIN CELL WITH STEM CELL DIVERSITY? Life Science Training from Industry Experts

INDUCED PLURIPOTENT STEM TERM OF THE WEEK: INDUCED

CELLS SHOW PROMISE PLURIPOTENT STEM CELL
Imagine being able to reprogram one of your own skin An induced pluripotent stem cell (IPSC) is a type of
cells to produce a functioning nerve cell or section of pluripotent stem cell that can be generated directly from
cardiac tissue. This may sound like science fiction — but adult cells. In theory, this means that a scientist could
the groundwork for this to become a reality is already in create a stem cell treatment using cells from a patient’s
the works as researchers expand their ability to create own body by following these steps:
and manipulate induced pluripotent stem cells. • Remove a skin or other easily accessible cell type
In this WEEKLY, we’ll get the details on what these from the patient.
multifaceted cells are all about and discover their • Manipulate the cell in the lab to produce an induced
therapeutic potential. pluripotent stem cell.

STEM CELL PRIMER • Add the required growth and differentiation

factors to get the induced pluripotent stem cell to
Stem cells are unspecialized cells that have the ability
differentiate into the desired tissue type.
to develop (differentiate) into 1 of 200 cell types in the
body. There are two general classifications: This newly differentiated tissue could then be
transplanted back into the patient’s body without fear of
• Embryonic stem cells (found only in developing
immune rejection since it is derived from their own cells!
embryos) can become any cell type within the adult
body. These are pluripotent stem cells. AN EASIER RECIPE
• Adult stem cells (found in the organs of an adult) The development of IPSCs was so significant that the
can only become certain cell types. Typically, these scientists at Cambridge University (Cambridge, UK) and
cell types come from the organ in which they are Kyoto University (Kyoto, Japan) who figured out how to
derived from. create them received the 2012 Nobel Prize in Medicine.
Due to their ability to differentiate (change into) into any Their discovery was based the observation that when
cell type in the adult body, pluripotent stem cells show pluripotent stem cells differentiate into specialized cell
the most promise as a therapeutic. The idea is they can types, certain genes are deactivated or switched off.
be induced in the lab to form a specific type of adult They wondered if the reverse might also be true — if by
tissue and then transplanted into a patient who needs reactivating or turning on those same genes, they could
that tissue. arrive at pluripotency. Through a series of experiments,
For example, someone who damages their spinal cord they hit upon the correct combination of genes to
may potentially benefit from a transplant of replacement reactivate, and succeeded in inducing pluripotency.
spinal cord tissue. In fact, these types of clinical trials
are currently ongoing and show promise. However,
RESEARCH APPLICATIONS
transplanting tissue derived from an embryo carries the The best way to decipher a disease is to examine the
same risks as an organ transplant — rejection by the affected cells from a patient — for example, a blood
immune system. Thus, patients undergoing this type of sample from a leukemia patient or a tumor biopsy from
therapy must receive immunosuppressive drugs, which a breast cancer patient. What if the affected tissue is
carries its own set of risks. impossible or dangerous to access — such as brain or
cardiac tissue? IPSCs to the rescue! To better understand
What could be a possible alternative that solves the
Alzheimer’s, researchers are creating IPSCs from
rejection issue?
Alzheimer’s patients’ skin cells, which are then induced
to become brain cells. In this manner, the disease models

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reflect the genetics of an Alzheimer’s patient without the develop IPSC-derived heart muscle cells and neurons
need to directly access their brain cells. These patient- for drug discovery applications.
based disease models are being used as both a way to
better understand the disease process as well as for CLINICAL APPLICATIONS
drug discovery research. Ultimately, scientists and physicians want to use patient-
Companies developing IPSC-based drug discovery derived IPSCs for individualized treatments as described
platforms include: above. Currently, there is only one ongoing clinical
trial using IPSCs, taking place at the RIKEN Institute
• Evotek’s (Hamburg, Germany) collaboration with
(Wako, Japan) for the treatment of wet age-related
Celgene (Summit, NJ) to use their IPSC platform
macular degeneration. The scientific community is
to discover and develop new drugs for a range of
closely watching this trial — if IPSC treatments prove to
neurodegenerative disorders, including Alzheimer’s,
be safe and effective, they will revolutionize an already
Parkinson’s, and ALS.
revolutionary field.
• Axiogenesis’ (Cologne, Germany) collaborations
with Metrion Biosciences (Cambridge, UK) to

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CELL SIGNALING EXPLAINED Life Science Training from Industry Experts

UNDERSTANDING THE
DISEASE PROCESS
Greetings from BIO 2017! It’s been a busy week here at
the BIO International Convention here in San Diego,
CA. At our convention booth, this year’s giveaway
was our book written especially for non-scientists:
The Biotech Primer: An insider’s guide to the biotech and
pharma industry. If you weren’t at the convention to
stop by and get a copy, please enjoy this excerpt below
on cellular communication – a topic fundamental to
understanding both normal and diseased cellular
processes, and a process that is modulated by many
drugs on the market today.
Another class of membrane proteins that aid in cellular
communication is channel proteins. These proteins
CELLULAR COMMUNICATION
act as molecular gates that allow the passage of small
In multicellular organisms, cells must communicate molecules and ions, for example, glucose and sodium,
with each other. Since cells don’t have mouths, ears, or across the membrane in response to a stimulus, such
access to email, they must rely on chemical messengers. as an electrical current in the case of ions or insulin
A chemical message – for example, a hormone – is signaling in the case of glucose. In neurons, ion transport
released by one cell, and received by a second cell – the between cells serves as a principle means of signal
target cell. The target cell receives the message through transduction. The influx of calcium ions (Ca++) into a
proteins inserted into its membrane known as receptors neuron results in the release of neurotransmitters –
– proteins that control the passage of molecules and the chemical messengers specific to the nervous system.
flow of information across the membrane. When the Different types of neurotransmitters regulate a variety
signaling protein binds its receptor, the receptor changes of brain functions, including muscular activity, memory
shape and transduces (converts from one form to and learning, and mood regulation.
another) the chemical message across the membrane to
the cell interior. This process of cellular communication
is known as signal transduction. The most common
end result of signal transduction, and a key step in cell
decision making, is the switching on, or off, of protein
production – more commonly called gene expression.

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 Alternatively, the signal and target may be in close
proximity and the signal can be transported by diffusion
through the intracellular space. After receiving a signal,
the target cell responds in a manner that is determined
by the nature of the signal received.

GROWTH FACTOR SIGNALING

Growth factors are proteins that signal a cell to multiply.
For instance, epidermal growth factor (EGF) stimulates
the proliferation of skin cells during wound repair.
Cells are constantly exposed to many different growth
factors, and the particular ones they respond to depends
on their cell surface receptors. Skin cells, as well cells
covering the gut, lung and breast, have or express
The regulation of blood sugar levels by the receptors for epidermal growth factor (EGF), while nerve
protein hormone insulin is an example of cellular cells express receptors for nerve growth factor (NGF).
communication. After you eat, beta cells in your
pancreas sense increased blood glucose and respond by
releasing insulin into the bloodstream. Insulin molecules
attach to specific insulin receptors on muscle cells and,
in doing so, deliver a signal to the inside of the muscle
cell to send glucose channels to the membrane, resulting
in glucose uptake. In this way blood glucose levels are
kept constant.

CELL SIGNALING: A CLOSER LOOK

Some cells send signals while others receive signals, but
most cells do both. The signals are chemical hormones,
such as adrenaline, or proteins, such as insulin. They
are produced within specialized cells (the signaling cell) After receiving the initial growth factor signal, the
and released to find their target cells. The signal is often enzymatic activity of the internal portion of the growth
called a ligand. In some cases, the signaling cell and factor receptor is activated. The particular type of
target cell may be the same cell. The target cell may be activity switched on is protein kinase activity – or the
in direct contact with the signaling cell, or it may be in a ability to transfer a phosphate group from one molecule
different part of the body and receive a signal that has to another. These types of receptors are sometimes
been transported through the bloodstream. referred to as receptor tyrosine kinases (RTKs), because
they selectively transfer phosphate groups to the amino
acid tyrosine on the recipient protein. This transfer, in
turn, causes a slight shape change in the protein which
received the phosphate group, typically leading to the
activation of that protein’s own kinase activity. This
newly activated protein kinase then goes on to activate
yet another kinase protein, and so on, in what is referred
to as a signal transduction cascade. The last element in
this cascade to be phosphorylated is typically a protein
called a transcription factor. Once phosphorylated, the
transcription factor enters the nucleus, where it binds to
the DNA at a particular location, activating expression of
a specific gene.

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 Defects in the growth factor signaling process are
associated with different types of cancer. A major
challenge in oncology lies in understanding the
complex signaling pathways that trigger cell division
and determining what has gone wrong in each type of
cancer. Once these signaling pathways are understood,
it is possible to develop targeted therapies for the
particular cancer.

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BREAST CANCER SUBTYPES Life Science Training from Industry Experts

FOUR MOLECULAR on the surface of HER2+ breast cancer cells also signals
VARIANTS EXPLAINED the patient’s immune system to attack that cell.

Hearing your doctor utter the words HER2-positive, Kadcyla, also made by Genentech, is an antibody-drug
HR-positive, triple-negative, or BRCA mutation can be conjugate — a monoclonal antibody that delivers a
devastating — even for the most resilient person. Simply highly toxic drug directly to HER2+ breast cancer cells.
put, breast cancer is a complex disease. A diagnosis can Kadcyla binds the HER2 receptor like Herceptin, but
be derived from any combination of the factors listed also delivers a toxic payload (which is actually attached
above — or, none at all. to the monoclonal antibody). As a normal part of the
cell’s lifecycle, cell-surface receptors get internalized or
The National Cancer Institute (Bethesda, MD) has
“taken up” by the cell on a regular basis. When Kadcyla
outlined four molecular subtypes of the disease.
is attached to a receptor that gets internalized, the
Each subtype is categorized by the cancer’s hormone
toxic payload is released from the antibody and kills the
receptor (HR) status and the level of expression from
cancer cell internally.
the HER2 gene. These cellular distinctions lead patients
on different treatment journeys because the cancer HR-POSITIVE
subtype determines the drugs used in a treatment plan.
About 70% of breast cancer diagnoses involve a
In this WEEKLY, we present a quick primer on the science significant number of receptors for either estrogen or
behind HER2-positive, HR-positive, triple-negative, and progesterone, making them hormone receptor positive
the BRCA gene. (HR+). HR+ cancers may respond positively to treatments
that block either the action or the production of
HER2-POSITIVE estrogen. In some cases, these treatments may continue
HER2-positive (HER2+) breast cancer patients — about to be used for up to five years after initial treatment in
20% of all breast cancer cases — have the most highly order to prevent recurrence.
effective therapies available on the market. HER2+
Two common types of medication for HR-positive breast
cancer cells produce, and therefore present, larger than
cancers are Tamoxifen and aromatase inhibitors. Both
normal numbers of the HER2 receptors on their cell
types of drugs may also be prescribed as a preventive
surface. These HER2 receptors capture growth factors,
treatment in women who are at high risk for breast
which trigger the cell to grow and reproduce more
cancer. In fact, Tamoxifen is named on the World
rapidly than normal. Mutations are more likely with rapid
Health Organization’s List of Essential Medicines, a list
reproduction and thus, a tumor is born.
of the most important medications needed in a basic
The overexpression of the HER2 receptor is a result healthcare system.
of having extra copies of the HER2 gene — known in
Tamoxifen works by inhibiting the estrogen receptor
the world of genomics as gene amplification. Gene
and was originally discovered by AstraZeneca (London,
amplification events are thought to be caused by
U.K.). On the other hand, aromatase inhibitors block
mutations that occur after a person is born — it is not an
the production of estrogen by inhibiting an enzyme
inherited form of cancer.
whose activity is required for estrogen production. The
Genentech’s (South San Francisco, CA) Herceptin is different aromatase inhibitors on the market include
a monoclonal antibody that binds to and blocks the Arimidex (AstraZeneca), Femara (Novartis; Basel,
activity of the HER2 receptor on cancer cells. When the Switzerland, and Aromasin (Pfizer; New York, NY).
HER2 receptor is blocked, the HER2 growth factor can no
Selective estrogen receptor degraders (SERDs) are drugs
longer bind and send a growth signal to the cell, so the
that bind to estrogen receptors and cause them to be
cancer cells stop dividing. The presence of an antibody
degraded. Fewer estrogen receptors mean that the cells
receive growth signals from estrogen. Currently, there is

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only one selective estrogen receptor degrader approved Mutated BRCA1/2 genes are non-functioning, so they
— Faslodex, marketed by AstraZeneca. A second SERD, cannot locate DNA damage, nor can they enlist DNA
Elacestrant, is in Phase I clinical testing by Radius Health repair. Testing positive for BRCA1/2 mutations may
(Waltham, MA). indicate there is an accumulation of DNA damage, which
Another new class of therapies for estrogen-receptor may eventually lead to cancer. BRCA is normally active in
positive breast cancer are small molecule inhibitors of breast and ovarian cells, which is why certain mutations
cellular enzymes known as cyclin-dependent kinases in BRCA1/2 are associated with a significantly increased
(CDKs). CDKs promote the development and division risk of developing breast or ovarian cancer. It must be
of cancer cells and inhibiting CDKs help to arrest stressed that BRCA1/2 mutations in and of themselves
cancer growth. do not cause cancer; they simply make it more likely
to occur.
The first CDK inhibitor, Ibrance (Pfizer) was approved in
2015. Kisqali (Novartis) was approved in March of 2017, A new class of drugs known as PARP1 inhibitors gives
and Eli Lilly’s (Indianapolis, Indiana) Abemaciclib is in hope to women whose breast cancer is associated
Phase III clinical development. with non-functioning BRCA genes. PARP1 is a second
type of DNA repair protein. By inhibiting this pathway,
TRIPLE-NEGATIVE DNA damage becomes so extensive that the cancer
cells commit “cell suicide” (or apoptosis.) When the
Triple-negative breast cancers lack receptors — they
cell in question is a cancerous cell, apoptosis is a very
are estrogen-receptor negative, progesterone-receptor
good outcome.
negative, and HER2-negative. Since there are no receptor
drug targets, this subtype is challenging to treat and The first FDA approved PARP1 inhibitor drug, Lynparza
there are no targeted therapeutics to date. If detected (AstraZeneca) was approved for BRCA associated ovarian
early enough, triple-negative breast cancer may respond cancer in December 2014. Clovis Oncology (Boulder, CO)
well to chemotherapy. achieved the second FDA-approval of a PARP1 inhibitor,
Rubraca, in December 2016, and Tesaro (Waltham, MA)
THE BRCA GENE garnered the most recent approval in March 2017.

BRCA stands for “BReast CAncer susceptibility gene” Not all triple-negative breast cancers are BRCA
and everyone has the BRCA 1 and BRCA 2 genes. The job associated, but many BRCA associated cancers are triple-
of BRCA is to scan cellular DNA for damage and trigger negative. For this reason, triple-negative breast cancer
DNA repair processes when mutations are found. BRCA patients may find hope in PARP1 inhibitor drugs.
genes are passed down from one generation to the next Breast cancer is a complex disease, and a better
— a good thing, unless the version passed down is a understanding of its molecular causes has enabled
mutated variation. researchers to develop more effective therapies. As our
understanding of the disease continues, we can expect
to see additional novel therapeutics.

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STOPPING A BIG PROBLEM: BLOOD CLOTS Life Science Training from Industry Experts

MEDS FOR THINNER BLOOD CAN VTE occurs when blood clots form in the absence of
EQUAL FEWER CLOTS, BUT HOW? an injury. These clots may break free and migrate to
another part of the body, where it may interfere with
The FDA’s recent approval of Portola Pharmaceuticals’
blood circulation and impair organ function. If this
(South San Francisco, CA) new blood thinner drug
occurs in a major organ such as the lungs, brain, or
Bevyxxa paved the way for the prevention of blood clots
heart, critical injury or death may result. The clots may
in patients hospitalized for conditions such as heart
also grow to a size large enough to block the flow of
failure, stroke, and pulmonary disease. The medical term
blood in the blood vessel in which it originally developed.
for blood clot is venous thromboembolism (VTE), but if we
Risk factors for VTE may be acquired (including older age,
take it apart:
major surgery, prolonged immobilization, certain type
• ”venous” means relating to a vein or the veins. of cancers, pregnancy and hormonal contraceptives)
• ”thrombo” is a blood clot. or inherited.

• ”embolism” involves the lodging of an embolus,

MECHANISM OF ACTION: BEVYXXA
a blockage-causing piece of material, inside of a
blood vessel. Bevyxxa and other drugs that prevent the formation
of blood clots belong to a class of drugs called
Hospitalized patients are at high risk for VTE because
anticoagulants which thin the blood. Bevyxxa works
of their restricted mobility — not being able to move
by directly inhibiting one of the key clotting factors,
causes blood to pool and collect in the body. An
Factor Xa. This differs from older anticoagulants such
especially dangerous type of VTE is deep vein thrombosis
as warfarin that works by inhibiting Vitamin K, which is
— blockage of a vein that is deep within the body, as
required for complete activation of clotting factors.
opposed to near the surface of the body. If a portion of
a deep vein thrombosis breaks off, it may travel to the Some key benefits of direct Factor Xa inhibition include
lungs, causing a potentially fatal pulmonary embolism. faster onset, less interaction with other medicines or
certain foods, and fewer bleeding events observed
An estimated 24 million people are hospitalized annually
during clinical trials, leading to a better safety profile.
due to VTE, so let’s find out how blood clotting is
Bevyxxa is the first oral Factor Xa inhibitor to be
activated and learn the science of Bevyxxa.
approved, and has been approved for use for up to 42
days. These attributes mean Bevyxxa can be prescribed
A CLOT IN THE DARK to a patient to continue taking the anticoagulant after
In healthy people, blood clotting is activated when release from the hospital.
tissue or a blood vessel is damaged, and involves
Patients were selected for treatment with Bevyxxa
specialized blood cells known as platelets — also known
based on increased levels of “D-dimers” in their blood.
as thrombocytes. Either type of aforementioned damage
D-dimers are degradation products of fibrin, the key
results in activating platelets, which then form an initial
protein component of blood clots. When our body
“plug” at the site of injury. At the same time, proteins
breaks down blood clots, D-dimers are produced. Thus,
known as clotting factors are also activated. Clotting
having higher than normal blood levels of D-dimers is a
factors work together to produce a protein called fibrin,
sign that higher levels of blood clots are present.
which is a fiber-like protein that forms a network of
strands that, together with the platelets, form a clot
AN ANTICOAGULANT U-TURN
at the site of injury. Clot formation in response to
injury prevents excessive bleeding and enables healing Anticoagulant drugs can be life-saving; however
to begin. their inhibitory effects may need to be reversed due
to major bleeding, or in the case of an emergency
surgery. Portola’s andexanet alfa, currently in late stage

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development, reverses Factor Xa inhibition. Andexanet Foods that we eat may also impact blood clotting ability.
alfa works by irreversibly binding Bevyxxa, preventing For example, foods high in vitamin E such as almonds
it from binding clotting Factor Xa. If Bevyxxa can’t and hazelnuts, as well as spices such as cayenne pepper,
interact with and inhibit Factor Xa, it no longer prevents garlic, ginger, and onion have some natural anticoagulant
blood clotting. effects, while foods high in vitamin K such as leafy
green vegetables, egg yolk, and soybeans may promote
COCKTAIL FODDER: BLOOD coagulation. For most healthy people, the relatively
THINNERS IN THE WILD small amounts of these foods consumed in a normal diet
Ever wonder how mosquitos and ticks are able to keep would not have a significant impact on blood clotting;
the blood flowing from their point of attack until they’ve however, those on anticoagulant medicines may want to
had their fill? It turns out that their saliva contains a consult their physician about any possible dietary impact
natural anticoagulant which prevents platelets from on their medicine’s efficacy.
being activated. Fortunately, the effect is only temporary
and localized to the site of the insect bite.

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UNPACKING DIGITAL MEDICINE Life Science Training from Industry Experts

CAN APPS PROGRAM BETTER HEALTH? DIABETES

Digital medicine is defined by the field’s pioneer Dr. Eric Diabetes management is a prime target for digital
Tool of the Scripps Translational Science Institute medicine intervention, as the number of people
(La Jolla, CA) as “the ability to digitize human beings, diagnosed with the disease has quadrupled over the
by a variety of means (sequencing, sensors, imaging, past 35 years, from 5 million to 20 million (Centers for
etc.), fully exploiting our digital infrastructure of ever- Disease Control).
increasing bandwidth, connectivity, social networking, WellDoc’s (Columbia, MD) BlueStar smartphone-based
the Internet of all things, and health information app is the first FDA-cleared mobile prescription therapy.
systems.” This new field is changing the way diseases The app allows users to enter data including levels of
from diabetes to substance abuse are prevented blood glucose, carbohydrates consumed, medications
and treated. taken, exercise amount, hours of sleep, and perceived
“Digitizing human beings” may sound impersonal — but stress levels. BlueStar then makes personalized
in fact the opposite is true. By enabling better access recommendations regarding diet, exercise, and
to individual health data, patients and physicians can medication, and even pinpoints the best times of day for
create truly personalized health management plans. In the patient to test blood glucose levels. This information
this WEEKLY, we’ll take a look at this emerging biotech is easily shared with a physician.
sector and the companies leading the way into the land In clinical testing, patients assigned the BlueStar
of digital medicine. app showed an average 1.9% drop in glycosylated
hemoglobin (HbA1C) levels when compared to patients
THERE’S AN APP FOR THAT treated according to the current standard of care and
There is a wealth of potential to delay the onset of no continuous intervention app. HbA1C is a reflection of
diseases confronting our population, and a number average blood glucose levels over the past three months.
of companies are developing digital medicine apps — Patients must receive a doctor’s prescription for the
similar to the ones you have already downloaded onto BlueStar app.
your phone such as your favorite music streaming app.
Another leader in digital therapeutics diabetes
Most of today’s digital medicine falls under the heading
management is Omada Health (San Francisco, CA),
of “medication augmentation” — interventions meant to
whose interactive behavioral intervention program
supplement rather than replace medication.
reduces the development of Type 2 diabetes in
Chronic diseases such as diabetes can be managed much prediabetics through personal coaching via the
more effectively with “continuous intervention” — a integration of web, mobile, and smart devices. The goal
day-to-day monitoring of the patient’s lifestyle choices is to help patients lose weight and increase physical
and medication compliance. In many cases, these apps activity. Although not FDA-approved, the platform is
are highly sophisticated, clinically validated, and FDA- recognized by the Centers for Disease Control as an
approved. The disease areas most commonly being effective diabetes prevention tool.
tackled include Type 2 diabetes, chronic respiratory
disease, chronic cardiovascular conditions, and mental CHRONIC RESPIRATORY DISEASE
health conditions. In some cases, digital health companies are partnering
Let’s unpack some of the digital medicine coming to an with pharma companies to ensure better use of a
app store near you. therapeutic drug. Propeller Health (Madison, WI) has
joined forces with GlaxoSmithKline for a “digitally
guided therapy” platform for use with inhalers to treat

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asthma and chronic obstructive pulmonary disease ADDICTION & SLEEP
(COPD).
Pear Therapeutics (Boston, MA) is tackling substance
The platform consists of sensors, provided by Propeller, abuse disorders with its digital therapy. Their lead
that are attached to patients’ inhalers; a smartphone product, reSET, is a smartphone app for patients
app for patient use; a website for physician use and with a clinician-facing web interface. It is designed to
data from a network of air-quality sensors. Propeller deliver behavioral therapy through a series of learning
monitors inhaler use, tracking patients over time and modules. The goal is to keep patients interested and
providing data on disease management as it relates to engaged in their treatment between therapist visits. The
environmental factors (air quality). The Propeller system apps have been clinically tested in five different trials.
has four FDA clearances which allow the company to Patients who received reSET treatment in addition to
claim the system can be used to increase medication standard addiction therapy showed better rates of drug
adherence, predict exacerbations, and reduce the abstinence. The app has been submitted to the FDA
frequency of symptoms and exacerbations in asthma for approval.
and COPD.
Big Health (San Francisco, CA) has developed a digital
HEART DISEASE therapy, Sleepio, to help with insomnia. The app consists
of cognitive behavior therapy (CBT)-based exercises
No discussion of digital medicine would be complete
delivered by an online, animated therapist dubbed
without a nod to the legendary story of Dr. Eric
The Prof. When tested against an online version that
Topol using AliveCor’s (Mountain View, CA) mobile
included interaction with the Prof but lacked CBT-based
electrocardiography (ECG) device and app to diagnose
activities, Sleepio was more effective at helping 75% of
a heart attack mid-flight. Dr. Topol was actually using a
the participants to fall asleep.
prototype model during his in-flight diagnosis; today, the
Kardia Mobile device — essentially two sensor pads — is THE FUTURE
FDA-cleared and available for $99. After downloading the
The companies and apps described here are really
accompanying smartphone app, users can get an ECG
just the tip of the iceberg of this new therapeutic
reading in 30 seconds by opening the app on a phone
world. Additional disorders for which digital therapies
placed nearby and placing their fingertips on the sensor
are being developed include obesity, hypertension,
pads. An irregular reading indicates possible atrial
hyperlipidemia, smoking cessation, chronic pain,
fibrillation, potentially indicating a heart attack.
coronary artery disease, and even serious mental
illnesses such as schizophrenia. The field is still in its
infancy, but the dramatic benefit already seen by many
adopters suggests a bright future.

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CHAPERONING THE RARE DISEASE DANCE Life Science Training from Industry Experts

PROPERLY FOLDING MISFOLDED Production of functional galactosidase enzyme is limited

DISEASE PROTEINS because of mutations in the galactosidase gene that
cause the enzyme to be misfolded and therefore non-
Amicus Therapeutics (Cranbury, NJ) found itself in
functional.
the news earlier this month when the FDA agreed to
review the company’s new drug application for their The only Fabry’s disease treatment on the market in
investigational therapy to treat Fabry’s disease. The the U.S. is Fabrazyme, which is made by Genzyme
drug under consideration, migalastat, has already (Cambridge, MA). Fabrazyme is an enzyme-replacement
been approved by the European Medicines Agency. therapy: since the patients don’t make enough functional
It belongs to a small, but growing class of therapeutics galactosidase enzyme, scientists produce it in the lab
known as pharmacological chaperones that properly fold using cells that have been genetically engineered to
improperly folded proteins that cause disease. produce the enzyme, which is then purified and injected
into patients.
Let’s take a look at which chaperones are on the dance
floor and find out the steps they are taking to treat MECHANISM OF ACTION: MIGALASTAT
disease caused by proteins.
Amicus’ drug migalastat, if approved, would be the first
TERM OF THE WEEK: small molecule treatment for Fabry’s. The potential
CHAPERONE PROTEIN availability of swallowing a drug (vs. injecting) would give
those with Fabry’s another drug delivery option.
Chaperone proteins are proteins that assist in the correct
folding and assembly of other proteins. Many of the In the lab, migalastat binds to and inhibits galactosidase.
proteins produced by our cells require chaperone In the body, this high affinity is taken advantage of by
proteins to ensure their correct molecular structure. migalastat binding to mutated galactosidase during
the process of folding, where it then shifts the folding
A pharmacological chaperone is a small molecule drug
towards the correct conformation. The now correctly
that targets specific misfolded proteins and encourages
folded protein makes its way to a cellular compartment
them to fold correctly.
known as the lysosome, where it carries out its job of
Protein misfolding plays a role in many different rare digesting lipids. The inside of the lysosome has an acidic
diseases, including enzyme deficiencies like Fabry’s pH, which causes migalastat to disassociate, leaving
and the related Niemann-Pick disease, as well as behind a functional galactosidase enzyme for the body
Huntington’s disease, and some cases of amyotrophic to pick up and use.
lateral sclerosis (ALS). Some of the mutations in the
Fabry’s is caused by a variety of different mutations
genetic disease cystic fibrosis (CF) involve misfolded
within the galactosidase gene; not all of them are
proteins. Diseases caused by misfolded proteins
amenable to treatment with migalastat. Amicus
that disrupt cellular function are sometimes called
scientists estimate that between 35% to 50% of patients
proteopathies, where proteo = protein, pathy = disease.
will be responsive to migalastat.

MECHANISM OF ACTION: FABRAZYME MECHANISM OF ACTION: LUMACAFTOR

A type of lysosomal storage disorder, Fabry’s disease
Another disease that can be traced to protein misfolding
involves the inability to process certain types of lipids
is cystic fibrosis. CF is a genetic disease caused by one
(fats), because they lack functional versions of critical
of several possible mutations in the gene encoding the
enzymes, resulting in a range of symptoms, including
“cystic fibrosis transmembrane conductance regulator”
kidney, heart, and skin disorders. The enzyme in
(CTFR) protein. The CTFR protein is critical for the
question here, galactosidase, helps to break down
production of sweat, digestive fluids, and mucus.
glycolipids — lipids with a carbohydrate attached.

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The most common mutation, responsible for about inclusion body myositis (sIBM), a rare muscular atrophy
two-thirds of CF cases, results in a protein that is so disease; and Niemann-Pick disease, a lysosomal storage
misfolded, it never makes it to the cell surface where it is disorder similar to Fabry’s disease.
required to do its job. Vertex Pharmaceuticals’ (Boston, Chaperone Therapeutics has a drug in preclinical
MA) drug lumacaftor serves as a pharmacological development for Huntington’s disease, which
chaperone for these proteins, assisting them with is associated with disordered folding of the
correct folding so that they can make it to the cell huntingtin protein.
surface. Lumacaftor is one piece of the CF puzzle; it is
often used in combination with other therapies to fight COCKTAIL FODDER: SHOCKING
various aspects of the disease. THE CHAPERONE
The largest family of naturally-occurring chaperone
MORE DANCE CHAPERONES
proteins are called “heat shock proteins” because they
Instead of creating pharmacological chaperones, were first discovered as part of a cellular response
another approach to getting mutated proteins to fold to heat shock — exposure to a higher than normal
correctly is to stimulate diseased cells to produce greater temperature. These proteins were later discovered to
amounts of natural chaperone proteins. This can be be induced in response to other types of cellular stress
done by identifying small molecules that induce cells to such as ultraviolet light exposure or wound healing.
express heat shock proteins, a common class of cellular It’s thought that these cellular stressors can disrupt
chaperones (described below). Two companies following protein folding, and the production of heat shock protein
this approach are Orphazyme (Copenhagen, Denmark) chaperones can help to counteract the disruption.
and Chaperone Therapeutics (Research Triangle Park,
Pharmacological chaperones that activate or mimic
NC).
these protective proteins may prove to be the fresh new
Orphazyme’s lead product, arimoclomal, has completed approach that can make a difference in a whole range of
Phase II clinical testing for amyotrophic lateral different diseases.
sclerosis (ALS) associated with mutations in the gene
for superoxide dismutase 1 (SOD1) enzyme; sporadic

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PUTTING THE CAR-T BEFORE THE HORSE Life Science Training from Industry Experts

THE STORY BEHIND CAR-T WHY CAR-T?

The hottest cancer therapy in the pipeline — chimeric In theory our immune system should recognize the
antigen receptor therapy (CAR-T) — got a big boost unique proteins presented on all diseased/cancerous
last month when an FDA advisory panel unanimously cells; however there are two main reasons this doesn’t
recommended approval of the treatment for children always happen:
and young adults with a severe form of leukemia who • Early on in the tumor development, the cell
have run out of other options. Developed by Novartis composition is similar enough to healthy tissue that
(Basel, Switzerland), this elegant hack of the immune it can be overlooked by the immune system.
system is one of many horses in the race for a FDA
• Later as a tumor progresses, it releases chemical
approval, with Kite Pharmaceuticals (Los Angeles, CA)
signals that suppress the immune response, helping
and Juno Therapeutics (Seattle, WA) rounding out the
it to evade detection. This trickery is known as
pack. Let’s take a moment to review these revolutionary
the tumor microenvironment and once again the
therapeutics and understand how they attack cancer.
dangerous cancer cells can pass by undetected.
TERM OF THE WEEK: KILLER T-CELLS So what’s a scientist to do?! Figure out a way to train
CAR-T therapy is modeled after a cell in the immune killer T-cells to ALWAYS recognize and destroy cancer
system known as the killer T-cell. The job of a killer T-cell cells… enter CAR-T.
is exactly what the name implies — to kill dangerous
cells. They target diseased cells in the body via their
HOW TO TRAIN AN IMMUNE SYSTEM
receptors: each one has a uniquely shaped receptor, Killer T-cells are “trained” to go after early and late stage
and will recognize its intended target because the shape cancer by having their physical structure altered. This
of its receptor “matches” or fits into a uniquely shaped alteration is accomplished by fusing an antibody with the
surface protein found only on diseased cells. Once the receptor of a killer T-cell to create a chimeric molecule —
Killer T-cell “docks” onto its target, it injects an enzyme or the “C” in CAR-T.
which triggers death. The result: no more bad cells. Training day begins by having killer T-cells drawn out of
a patient’s body and isolated in the lab. Next, scientists
deliver a gene to the T-cells that encodes the chimeric
receptor. This receptor consists of two parts:
• A targeting domain: This is the part of the chimeric
receptor that will be outside of the T-cell. It is
composed of an antibody that will recognize
and dock onto a unique surface protein of the
patient’s cancer.
• An activation domain: This part of the receptor will be
triggered once the targeting domain is engaged. It
will signal to the killer T-cell to:
1. Stay alive.
2. Make copies of itself.
3. Release signaling molecules called cytokines.
Cytokines are chemical signals that activate
other white blood cells to join the fight against
the tumor.

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 4. Kill the target cell.

WHAT’S IN A NAME?
The T-cell/antibody hybrid is now a CAR-T therapeutic.
Chimeric antigen receptor therapy broken down:
It is then multiplied in the lab and infused back into the
patient’s body. Once inside, the CAR-T locks onto its • Chimeric: Composed of components from two
cancer target, replicates, sends out cytokines, and kills distinct parts, such as an antibody and a killer T-cell.
the designated cancer cells. The CAR-T will continue • Antigen: A protein that is recognized by an antibody,
to replicate and kill any and all cancer cells recognized such as a protein on the surface of a tumor cell.
by the initial antibody component, with the goal of
• Receptor: A protein that is embedded in a cell
eliminating the disease.
membrane and transmits signals to itself in response
to being activated, for example a T-cell receptor
transmits signals to the T-cell when it docks onto
its target.
• Therapy: A treatment meant to manage or cure
a disease.
As these therapies begin to move from clinical trials
into clinical practice, the treatment of cancer will truly
be revolutionized, offering new hope to patients and
their families.

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THE SCIENCE OF CRISPR/CAS9 Life Science Training from Industry Experts

CRISPR/CAS9 UPDATE for an enzyme that cut DNA. This enzyme became known
as Cas, short for “CRISPR-associated”.
As CRISPR/Cas9 adds new indications to its resume,
legal battles over its IP continue to be waged in the US In the mid-2000’s, scientists realized the “spacers”
and Europe. matched DNA sequences of invading viruses — the
bacteria were storing away bits of invading viral DNA
On the clinical front, CRISPR/Cas9 entered its first
between its own bacterial CRISPR sequences! These
human trial at Sichuan University (Chengdu, China) last
bits of viral DNA create a “genetic memory” of the virus,
fall for metastatic lung cancer, and is widely expected to
enabling the bacteria to fight back if reinfected.
do so in the U.S. by the end of the year. This past March,
a team of scientists at Oregon Health and Science Reinfection triggers the following steps:
University (Portland, OR) announced that they had • Viral DNA present in the spacer sequences is copied
successfully edited a gene linked to severe heart defects into viral RNA.
in human embryos. • The DNA-cutting enzyme Cas is made, and
On the patent front, CRISPR technology and its attaches itself to the viral RNA produced from the
applications were discovered by two different research spacer sequence.
teams, one at University of California, Berkeley, and • This newly minted viral RNA/Cas complex finds its
another at the Broad Institute (Cambridge, MA). Both “match” on the invading viral DNA.
have filed patents on various aspects of the CRISPR/Cas9
• The Cas enzyme is now positioned to cut up viral
system. The Broad Institute had granted an exclusive
DNA, destroying the invading virus.
license to Editas Medicine (Cambridge, MA), while
Berkeley had granted licenses to Caribou Biosciences
(Berkeley, CA), CRISPR Therapeutics (Basel, Switzerland
USE IN HUMANS
and Cambridge, MA), Intellia Therapeutics(Cambridge, In 2013, researchers adapted this bacterial defense for
MA), and ERS Genomics (Dublin, Ireland). In February, use in human cells. Human cells were engineered to
the U.S. Patent Office ruled in favor of the Broad contain both specially-designed RNA and Cas genes.
Institute and its licensee, while in March the European When these human cells produce the RNA/Cas complex,
Patent Office ruled in favor of U.C. Berkeley patents. the dynamic duo is ferried to its complementary DNA
The legal battle is certainly far from over. target. Once in position, Cas goes to work cutting the
DNA. The particular Cas protein chosen for this work was
With all of these new developments making waves in the
one discovered in Streptococcus bacteria, Cas9 — hence
industry, let’s review the basics.
the moniker CRISPR/Cas9.
CAS TO THE RESCUE The ability to cut human DNA in precise locations is an
exciting innovation because of what the cell does next.
CRISPR was originally discovered as a key component of
the bacterial immune response. Bacteria, like people, are
plagued by viral infections, and bacteria have evolved
BREAKING & FIXING
clever ways to attack invading viruses. In the 1980’s, Cas9 creates double-stranded breaks (DSB) in the
scientists observed an interesting pattern in bacterial specified DNA sequence. DSBs cut both strands of the
genomes: repeating, palindromic sequences, with unique double-stranded DNA helix. Think of DNA as a two-lane
sequences referred to as “spacers” between the repeats. bridge that, after experiencing an earthquake, has a
They dubbed these regions a tongue twister of a name, section break off and fall into the water below.
“clustered regularly interspaced short palindromic DSBs activate two repair pathways to fix the break in
repeats,” or CRISPR. Scientists also noticed CRISPR the DNA:
sequences were always located near a gene that coded

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 • Non-Homologous End-Joining (NHEJ) closes the gap The PD-1 gene produces the PD-1 protein, which is
between the break by joining the two sections located on the T-cell’s surface. When the PD-1 protein
back together—visualize pushing the two sides of is activated, the T-cell doesn’t function. When the PD-1
the bridge together, leaving the fallen section in protein is deactivated, the T-cell functions. Aggressive
the water. An unintended byproduct of NHEJ is the cancers take advantage of this on/off switch turning PD-1
possibility of sequence error, much like the sections on, effectively shutting down the T-cell. By turning PD-1
of the bridge not lining up properly. If the repair off, the T-cells can’t be suppressed—freeing them up to
occurs in the middle of a gene, the minor error can attack cancer cells.
be enough to disrupt gene function and halt the
production of the corresponding protein. COMING SOON
• Homology Directed Repair (HDR) relies on a highly A U.S. clinical trial of CRISPR to disrupt PD-1 in T-cells is
similar (homologous) DNA segment to repair the expected to begin before the end of 2017. This two-year
break—visualize the missing bridge section built study is funded by the Parker Institute (San Francisco,
elsewhere and helicoptered in to fill the break. CA).

By engineering double-stranded breaks to occur at A number of private companies also have plans for
specific locations, scientists activate the NHEJ or HDR CRISPR/Cas9 clinical trials that include both gene
cell repair pathways. By activating the NHEJ pathway, disruption and gene correction. The table below
scientists can disrupt a disease-associated gene, summarizes some key players in the genome-editing
preventing the production of a protein that causes arena and their approaches to applying CRISPR. In vivo
the disease. By activating the HDR pathway, a short means the therapy will take place inside the human
sequence of DNA is delivered with CRISPR/Cas9 to and Ex vivo means the treatment will be performed in
correct the mutated sequence, perhaps allowing a cells taken from the body and then injected back into
missing protein that causes disease to be made. In both the patient.
scenarios cures for many different types of diseases may
be realized.

As these and other potential treatments move through

clinical trials, the world will be watching to see if this
revolutionary technology will live up to the hype and
CRISPR IN THE CLINIC change the way we prevent and treat disease.
A clinical trial for metastatic lung cancer, initiated last
fall by Chinese researchers at Sichuan University use
CRISPR/Cas9 to disable the PD-1 gene in T-cells.

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THE MICROBIOME MAGNIFIED Life Science Training from Industry Experts

DECODING THE GUT-BRAIN AXIS TRAIL BLAZERS: AXIAL &

There is no shortage of microbiome-focused startups THE MAZMANIAN LAB
in biotech right now. The link between the gut Axial Biotherapeutics (Cambridge, MA) is focusing
microbiome — the entire collection of microbes living much of its initial attention on work originally done at
in the gut — and diseases such as inflammatory bowel microbiologist Sarkis Mazmanian’s lab in the California
disease are well-established. New research has made Institute of Technology (Pasadena, CA) — mostly
it clear, however, that the gut microbiome also impacts relating to the gut-brain axis’ role in Parkinson’s disease
neurological health, leading to the phrase “the gut-brain (PD) and autism.
axis.” Let’s explore this connection and examine the Parkinson’s Disease
early efforts by a few innovative biotechs to translate
Parkinson’s disease (PD) is a chronic and progressive
these new discoveries into the clinic.
movement disorder, according to the Parkinson’s
GUT MICROBIOME PRIMER Disease Foundation. Symptoms include tremor of
the hands, arms, legs, jaw, and face; slowness of
The human microbiome is the complex collection of
movement; rigidity of the limbs and trunk; and impaired
microbes (mostly bacteria, but also includes small
balance and coordination. These symptoms are
numbers of fungi and viruses) that reside on and
caused by the malfunction and death of neurons that
inside the human body, including our skin, mouth,
produce the neurotransmitter dopamine. PD affects
nose, respiratory tract, and digestive tract (gut). The
nearly one million people in the U.S., and the cause is
microbiome is huge — microbial cells outnumber human
unknown. About 75% of PD cases are accompanied by
cells by a ten to one ratio! Human cells are much larger
gastrointestinal disorders such as constipation, which
than bacteria cells, however, so don’t worry — you’re still
provided an initial impetus to examine a possible
mostly human. For every 100 pounds that you weigh, it
connection between gut health and the disease.
is estimated that about two pounds of that weight come
from bacteria. A key molecular characteristic of PD is the aggregation
of a protein called alpha-synuclein (αSyn) within cells of
Most of us think of bacteria as harmful and certainly
the brain and gut. Researchers in the Mazmanian lab
some types are; however, those that have evolved with
used a strain of mice that overproduce αSyn to study
humans to become part of the human microbiome
the disease. One group of the αSyn mice were bred in
are either neutral — causing no harm — or beneficial.
a completely sterile environment to create “germ-free”
Scientists are busy trying to better understand and
(GF) αSyn mice. The other αSyn mice had the normal
characterize these beneficial bacteria and the role that
collection of gut bacteria. On a series of tests designed
they play in human health.
to assess motor skills, the GF αSyn mice performed
The gut contains the largest number of bacteria, significantly better — suggesting that even in mice that
as well as the greatest diversity of bacteria, when overproduced the αSyn protein, the presence of certain
compared to other parts of the body. Thus much of microbes are required for the disease to progress.
the attention directed towards the human microbiome Further work suggested that a molecule called butyrate,
has been focused on the gut microbiome in particular, produced by certain gut bacteria, can enter the brain
which continues to surprise us with its influence on and activate an immune response, leading neurons to
diseases such as obesity, diabetes, and, increasingly, malfunction or die.
brain disorders.
There is reason to believe that this connection is also
at work in humans with PD. In collaboration with Rush
University (Chicago, IL) gastroenterologists, Mazmanian
lab researchers transferred fecal samples from PD

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patients into the GF αSyn mice. Fecal transplants are lab demonstrated that feeding ASD mice a specific
an established way to “reset” the gut microbiome of strain of bacteria called B. fragilis — a part of the
the recipient to make it match that of the donor. After human microbiome — altered the mouse microbiome
transplantation, the mice began exhibiting symptoms of and reduced some of the ASD-like behaviors such as
PD. Transfer of fecal matter from healthy people did not anxiety and repetitive behavior, and increased levels
trigger these symptoms. These experiments suggest that of communication with other mice. These experiments
the gut microbiome is a major contributor to the disease suggest a possible probiotic therapy for autism.
process in PD patients.
Of course, these promising early stage findings still need
CARB LOADING WITH SYMBIOTIX
to be translated into human therapeutics. This may be Another early-stage company making headlines in
easier than traditional neurological approaches because the gut-brain axis space is Symbiotix (Boston, MA).
it is much easier to deliver drugs to the gut than to get Focusing on multiple sclerosis (MS), their lead candidate
them to cross the blood-brain barrier. Following up with is a carbohydrate molecule produced by B. fragilis. This
a targeted approach to modulate the production of therapy increases the production of regulatory T-cells,
butyrate and other inflammatory compounds produced which are a class of T-cells that “turn down” an overactive
in the gut may bring the first truly effective PD therapy immune response by releasing anti-inflammatory
into the clinics. signaling molecules. Symbiotix is preparing to enter
clinical trials with an orally-administered product for the
Autism Spectrum Disorder
treatment of MS and inflammatory bowel disease.
Autism spectrum disorder (ASD) is a developmental brain
The emerging work described here gives credence to the
disorder characterized by impaired social interaction,
old expression “think with your gut.” In addition to the
communication, and restrictive and repetitive behavior.
diseases discussed above, scientists are discovering links
These symptoms impact a child’s educational, social,
between the gut microbiome and other brain disorders
emotional, and physical development. More than 3.5
such as anxiety and depression. As the story continues
million Americans live with an autism spectrum disorder
to unfold, we are likely to see new therapeutics based on
according to the Autism Society. The cause is unknown,
restoring the balance that millions of years of human-
although genetics is thought to play a role.
microbe co-evolution has fine-tuned.
Similar to PD, a significant portion of ASD patients
exhibit gastrointestinal problems. The Mazmanian

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EYE OF THE CYTOKINE STORM Life Science Training from Industry Experts

THE FLASH OF THE FIRST CAR-T example, lung failure induced by excessive amounts of
fluids and cells moving into the lungs.
Last week’s much anticipated FDA approval of the first
chimeric antigen receptor T-cell (CAR-T) therapy for acute A cytokine storm is the adverse event most associated
lymphocytic leukemia hails as the first gene therapy on with CAR-T treatments and next generation CAR-T
the US market. treatments are being developed to have built-in controls
to regulate cytokines so the storms can be stopped.
Classified as a “cell-based gene therapy,” Novartis’
(Basel, Switzerland) Kymriah works by removing patients’ AN ON/OFF SWITCH
T-cells, using a viral vector to introduce a gene that will
In first generation CAR-T, maximum activation occurs.
allow the T-cells to recognize and kill cancer, and then
With a full cytokine barrage, there is no way to tamp
infusing these modified T-cells back into the patient.
down the cytokine response.
Recall T-cells are found in the blood and fight disease.
In next generation CAR-T, a small molecule drug may be
Along with its significant potential, Kymriah also carries
co-administrated with the therapy. The drug’s function
serious risks. Its approval came with a boxed warning
is to activate CAR-T to fight cancer, or turn it “on.” If a
because of the potential for “cytokine release syndrome
cytokine storm ensues, the small molecule drug can be
(CRS),” also referred to as a cytokine storm, which has
immediately withdrawn — the “off” switch — essentially
caused fatalities in clinical trials of other CAR-T products.
deactivating CAR-T and stopping cytokine release.
Making CAR-T safer while maintaining efficacy are goals
This second iteration of CAR-T is made possible by a
of next generation CAR-T. Let’s explore cytokine storms
handful of companies who are designing drugs that
and find out how scientists aim to circumvent this
will act as an “on/off switch” to control CAR-T. Bellicum
roadblock to fighting cancer.
Pharmaceuticals (Houston, TX) is developing a CAR-T
TERMS OF WEEK: CYTOKINE product, BPX-601, that uses small molecule-activation.
& CYTOKINE STORM BPX-601 entered Phase I clinical trials in February 2017.
Intrexon (Germantown, MD) has a similar product in
Cytokines are small proteins which play an important
Phase I clinical development.
role in relaying messages from one cell to surrounding
cells and tissue. Cytokines serve two main functions GETTING BISPECIFIC
involving white blood cells:
Juno Therapeutics (Seattle, WA) is developing a CAR-T
• Activate additional white blood cells to fight product that uses a sort of two-step verification process.
off pathogens It turns out that tumor cells have many proteins on their
• Stimulate white blood cells to move towards sites surface — so it is a challenge to find a distinct protein
of inflammation that is also unique to any given cancer cell. Instead of
Cytokine signaling makes for a very quick and strong relying upon finding that one special protein, why not
immune response. Usually, the response is kept in check, target a more common one and use another protein to
and dissipates when the bad cells have been eliminated. double check the work?

However, in some cases, this positive feedback loop — Juno’s approach: bispecific chimeric antigen receptors.
activated cells releasing still more activating cytokines This means each engineered killer T-cell has not one, but
— spins out of control, resulting in a cytokine storm. two chimeric antigen receptors (CARs).
Acute inflammation with accompanying symptoms such • One CAR is activated in the presence of a protein
as high fever, swelling, and nausea can occur. In severe found on the surface of cancer cells. Once activated
cases, serious tissue damage and death can result — for this CAR-T cell would produce more copies of itself,
release cytokines, and attack the tumor.

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 • A second CAR called an inhibitory CAR (iCAR), is the cancer cell, where it interacts with other proteins in a
activated in the presence of a different protein found cytokine signaling pathway.
only on healthy cells — NOT on cancer cells. If an Scientists at the Blood Research Institute of Blood
iCAR is activated, an inhibitory signal is sent to the Center of Wisconsin (Milwaukee, WI) have developed
first CAR, preventing the CAR-T from working. decoy molecules that interfere with the protein-protein
Simply put, one CAR finds the target protein while the interactions in the pathway. These decoy molecules are
other iCAR verifies the cell is “unhealthy” via another short protein fragments called peptides. By binding to
protein. This bispecific CAR-T, now in preclinical proteins in the cytokine pathway, the signal to produce
development, aims to eliminate the off-target effects more cytokines is blocked.
and decrease the amount of cytokines released.

The decoy peptides reduce cytokine production by

~70%, which is likely enough to prevent the immune
OXYGEN DEPRIVED response from spinning out of control. In the words of
Earlier this year, Cellectis (Paris, France) published a Laura Savatski of the Blood Research Institute, “By
paper describing work they’ve done to engineer CAR-Ts reducing cytokine production by CAR-T cells, you prevent
with an oxygen-sensitive domain. a cytokine storm from happening. So instead of dealing
Under normal cellular conditions, this domain signals the with a problem at the back end of the therapy, you solve
CAR-T to remain inactive. Under low oxygen, or hypoxic it at the front end through intelligent design.” Decoy
conditions, this domain sends an activation signal to molecules are currently in preclinical development.
the CAR-T. Since most solid tumors have a hypoxic With some of the best minds in the biopharma industry
environment, an oxygen-sensitive CAR-T should be working on CAR-T design, this landmark FDA approval is
activated within the tumor but not outside of it. likely to be just the first shot in a treatment revolution.

INTELLIGENT DESIGN THROUGH DECOYS

It turns out that CAR-T activation results in cytokine
production because part of the CAR-T is actually inside

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THE MULTIPLE MYELOMA LANDSCAPE Life Science Training from Industry Experts

BLOOD CANCER: MULTIPLE MYELOMA time, production of a substance that activates bone-
reabsorbing cells, osteoclasts, is increased. The resultant
Plasma cells are the antibody-producing cells of our
damage to the bone structure creates soft spots or
immune system which happen to play a critical role in
lesions which may extend from the inner bone marrow
our defense against infections. In multiple myeloma,
to the outside surface of the bone. Bone lesions result
plasma cells begin to grow and divide in an uncontrolled
in significant pain and increase the risk of fracture. Bone
manner, forming a cancerous mass known as a
destruction also releases excessive calcium into the
plasmacytoma. Marrow — which produces plasma — no
bloodstream, leading to a range of symptoms including
longer functions in our defense, it simply takes up space
confusion, nausea, and loss of appetite. Excess blood
inside the bone.
calcium, combined with high levels of M protein, also
What does biotech have in store to fight multiple contributes to impaired kidney function seen in multiple
myeloma? Let’s find out the treatments on the market myeloma patients.
and the up-and-comers in development.
UNMASKING MULTIPLE MYELOMA
EASILY CONFUSED: PLASMA There is no one diagnostic test for multiple myeloma.
CELLS VS BLOOD PLASMA Blood and urine tests to detect some of the symptoms
Plasma cells are specialized white blood cells that listed above such as low blood cell counts, elevated
produce infection-fighting antibody proteins. Most blood calcium levels, and impaired kidney function may
plasma cells are found in the bone marrow. suggest multiple myeloma. These tests are followed by a
Blood plasma is the straw-colored liquid component bone marrow biopsy for confirmation.
of blood that holds blood cells in suspension, made Most cases of multiple myeloma have no known cause,
up of water (95%), proteins, glucose, clotting factors, although some research suggests that regular exposure
electrolytes, hormones, carbon dioxide, and oxygen. to herbicides, insecticides, petroleum products, heavy
metals, and asbestos increases the risk of developing
PICKING APART PLASMACYTOMA the disease. And although there is not a specific gene
Plasmacytoma formation can lead to a host of yet associated with multiple myeloma, abnormalities in
problems with recognizable clinical symptoms. Instead chromosome structure or number are associated with
of producing normal disease-fighting antibodies, the disease.
plasmacytoma cells produce abnormal antibodies called
M proteins, which don’t provide any benefit to the body ON THE MARKET
and crowd out normally functioning antibodies. And Once considered incurable, there are now a number of
because all blood cells are formed in the bone marrow, effective treatments for multiple myeloma, and several
overproduction of plasma cells can also crowd out more are in the pipeline.
normal blood-forming cells. This can lead to anemia,
Darzalex ( Johnson & Johnson; New Brunswick, NJ)
caused by a shortage of oxygen-carrying red blood
and Empliciti (Bristol Myers Squibb; Princeton, NJ) are
cells; increased bruising and bleeding due to a reduction
both monoclonal antibody therapeutics approved to
in clot-promoting platelets; and an increased risk of
treat multiple myeloma. They work by recognizing and
infection due to lower levels of healthy infection-fighting
binding to proteins on the surface of multiple myeloma
white blood cells.
cells, activating the patient’s immune system to destroy
Although multiple myeloma is classified as a blood those cells.
cancer, it has a significant impact on bone health.
Ninlaro (Takeda; Osaka, Japan) is a small molecule
As the plasmacytoma grows, bone-forming cells
proteasome inhibitor therapy. A proteasome is a
called osteoblasts are suppressed. At the same
specialized compartment within the cell that gets rid of

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damaged proteins by digesting them. If the proteasome activity to these cells. Mivebresib is currently in Phase I
is inhibited, damaged proteins build up within the cell. clinical testing for multiple myeloma.
This triggers a process called apoptosis — essentially, Selinexor helps to increase the number of tumor
cell suicide. In other words, the cancer cell kills itself. suppressor proteins present in the nucleus of cancer
Farydak (Novartis; Basel, Switzerland) is a small cells. These proteins help to protect against cancer by
molecule “histone deacetylase (HDAC) inhibitor.” HDACs detecting DNA damage and promoting apoptosis in
are enzymes that modify chromosomes (strands of those cells that have high levels of DNA damage. In many
DNA that contain our genes) and influence how often types of cancer cells, tumor suppressor proteins are
specific genes are activated. Some cases of multiple transported out of the nucleus, where they can no longer
myeloma are associated with changes in gene activation. do their job of detecting DNA damage. By blocking this
By inhibiting HDACs, Farydak can correct this changed transport, Selinexor enables tumor suppressor proteins
gene expression. to do their job of triggering apoptosis is cancer cells.
Selinexor began Phase III clinical testing for myeloma in
IN THE PIPELINE June 2017.
Two novel drugs in the multiple myeloma pipeline are CAR-T therapies are also in development for multiple
Mivebresib (AbbVie; North Chicago, IL) and Selinexor myeloma. Bluebird Bio (Cambridge, MA), in partnership
(Karyopharm Therapeutics; Newton, MA). with Celgene (Summit, NJ), and Nanjing Legend Biotech
Similar to Farydak, Mivebresib influences the activation (Nanjing, China) have announced promising results in
of specific genes by inhibiting a group of proteins early phase CAR-T trials for multiple myeloma.
called Bromodomain and Extra Terminal motif (BET) Multiple myeloma is a complex cancer. In recent years,
proteins. In some types of cancer, genes are activated a better understanding of the disease has led to the
or deactivated inappropriately due to BET activity. By approval of several new therapeutics. In the coming
inhibiting BET, Mivebresib may restore normal gene years, we can look forward to additional approvals as
novel therapeutics move through the pipeline.

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FROM FANTASY TO REALITY:
XENOTRANSPLANTATION Life Science Training from Industry Experts

TRANSPLANTING ORGANS FROM • Determining the correct animal donor, taking into
ANIMALS INTO HUMANS account both anatomical and ethical considerations.

Every ten minutes, a new person is added to the national • Preventing immune rejection: Immunosuppressive
transplant waiting list. A little more than 75,000 people drugs need to be given at a higher dose for
are active waiting list candidates — meaning they are xenotransplant recipients than for those
medically eligible for transplantation according to the receiving organs from human donors. High
Organ Procurement and Transplantation Network. doses of immunosuppressive drugs can lead to
Over the past decade, the gap between organ supply and serious infections.
demand has continued to grow; an estimated 20 people • Lingering concerns that viral genes present within
a day die as they wait for an organ transplant. animal genomes, which do not harm the animal,
The idea of xenotransplantation — transplanting organs could be harmful to humans.
from animal donors to humans — has been the subject Today, pigs are considered to be the best potential
of discussion for decades in science circles. Now, with animal organ donor for humans, due to their availability
the tools provided by modern biotech, we may soon and the size of their organs closely matches human
see this seemingly science-fiction idea become a life- organ size. Let’s see what biotech is doing to address the
saving reality. Let’s explore the past, present, and future two remaining challenges.
of xenotransplantation.
THE FUTURE
THE PAST Cambridge, MA-based eGenesis is using CRISPR/Cas9
The dream of xenotransplantation has been around genome editing to tackle the problems of immune
for over a century. In 1905, a French scientist inserted rejection and potential interspecies virus transfer —
portions of a rabbit’s kidney into a child suffering from referred to as “porcine endogenous retroviruses (PERVs)”
kidney failure. The patient’s kidney function improved; in pigs. CRISPR can be used to “knockout” (remove)
however long-term follow-up was not possible as the undesirable genes, or to “knock-in” (add) desirable
child died of pneumonia within a few weeks. In the gene sequences.
early 1960s, a small group of critically ill patients at Last month, eGenesis scientists published a paper in
Tulane University received kidney transplants from which they described their successful use of CRISPR
chimpanzees. None of these patients survived long- genome editing combined with a cloning technique
term, and the establishment of working cadaver organ called “somatic cell nuclear transfer” to produce dozens
procurement programs put xenotransplantation on the of healthy pigs whose genomes no longer contain PERVs.
back burner until 1984 when “Baby Fae” captured the The pigs were produced using the following steps:
public’s imagination as the first infant to receive not
• Use CRISPR to inactivate PERV sequences present in
only a heart transplant, but a xenotransplant, when she
an adult pig’s DNA.
received a baboon heart. The transplant operation itself
was successful; however, the baboon heart was rejected • Transfer the modified DNA into a pig egg whose own
by Baby Fae’s immune system 21 days after surgery. nucleus has been removed. This technique is known
as somatic cell nuclear transfer.
THE PRESENT • The egg, now containing a PERV-free pig genome,
A few key challenges have prevented xenografts from is stimulated with an electric shock to trigger
solving the problem of organ shortages for patients in cell division.
need. They include:

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 • After several rounds of cell division, an early-stage eGenesis is not the only company using genome
embryo is formed, which can then be transferred editing in the pursuit of xenotransplants. Synthetic
into a surrogate mother to complete development. Genomics (La Jolla, CA), in collaboration with United
This was the first time that as many as 25 genes had Therapeutics (Silver Spring, MD), is also working on
been simultaneously and precisely deactivated with modifying pig genomes to make their organs safe for
CRISPR — a significant accomplishment and a major step human transplant.
towards making xenotransplants safe. We are still several years away from making
More work still needs to be done, however, before pig- xenotransplants a routine clinical reality. The next
to-human heart transplants are routine. In addition to step will be testing genetically modified pig hearts in
deleting PERV genes, scientists will need to deactivate baboons. But with two of the leading genomics pioneers
pig genes that help to trigger immune rejection and add behind eGenesis and Synthetic Genomic — George
in gene sequences to help the human immune system Church and Craig Ventor, respectively, this is certainly
recognize the transplant as safe. an area to keep close tabs on.

As for the cloned pigs, the next step will be to see if the COCKTAIL FODDER: ORGANS FOR SALE?
changes made to their genome will be transmitted to
The only place one may buy an organ legally is in the
subsequent generations because breeding pigs are much
country of Iran; however, citizenship is required in order
easier than cloning them. And speaking of pig breeding,
to lessen transplant tourism. Australia and Singapore
Smithfield Foods (Smithfield, VA) — better known for
have legalized monetary compensation for living organ
producing pork products for consumption — has thrown
donors, in order to cover associated medical expenses
its hat in the ring by establishing a bioscience unit to be
and compensate for time of work.
used for supplying pig parts for medical use, ultimately
including organs for xenotransplantation.

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PICTURING DISEASE Life Science Training from Industry Experts

USING MEDICAL IMAGING TO gamma rays, can be created, manipulated, and

INVESTIGATE DISEASE detected for various applications, including medical
imaging technology.
Medical imaging — using various modalities to take a
snapshot of the body’s interior structure — has been CT VS. PET
around since 1895, with the discovery of X-rays by
Medical X-rays, as described above, work by beaming
Wilhelm Roentgen.
electromagnetic radiation at the body from an outside
X-rays are a type of electromagnetic radiation (more on source (the X-ray machine). “Positron emission
that later!) that are able to pass through soft tissues such tomography” — a PET scan — in contrast, assesses
as skin, fat, and muscle — but not bone. When an X-ray bodily function from the inside out. A radioactive tracer
beam is aimed at the body, it passes through the soft is injected and seeps through the body and attaches
tissue but is blocked by the bone, casting a shadow to itself to certain tissues, then the gamma rays emitted by
create an X-ray image on a piece of film. The introduction the tracer are detected by the PET scan. The radioactive
of X-rays revolutionized medicine, making it possible tracer (also called a radiopharmaceutical) consists of a
to accurately diagnose broken bones and to identify “carrier molecule” that is bonded tightly to a radioactive
the location of harmful objects such as bullets inside of atom. The carrier molecule used has an affinity for the
patients’ bodies. part of the body to be imaged.
Newer technologies such as PET and CT scans CT scans — “computerized tomography” — are
supplement X-rays, aiding in the diagnosis and essentially 3D X-rays. X-rays are taken from many
management of a disease. Let’s find out how different angles, creating cross-sectional images or
these pictures are worth much, much more than a “slices.” These cross-sections are then processed by a
thousand words. computer to produce a 3D anatomical image. A contrast
agent — which is used to enhance the body’s structure
TERM OF THE WEEK: or fluids — is typically given to patients prior to a CT
ELECTROMAGNETIC SPECTRUM scan, making the soft tissues denser, therefore enabling
Electromagnetic radiation is a combination of electric the agent to block X-rays and thus become visible in
and magnetic fields from various wavelengths that move the CT.
through space and carry energy. The electromagnetic
This combination of both types of tomography — the
spectrum encompasses the following:
use of waves to penetrate the body to create images — is
very useful when information about both metabolism
(PET) and anatomy (CT) are required to study disease.

PARKINSON’S & ALZHEIMER’S

Radioactively labeled glucose — fluorodeoxyglucose
(FDG) — is widely used for metabolic imaging studies in
PET scans because they detect deficits in brain activity
that may show signs of neurodegenerative diseases such
as Alzheimer’s or Parkinson’s.
Since all types of tissue use glucose for energy, increased
uptake of glucose-loving FDG indicates more metabolic
We can only see a small portion of the electromagnetic activity. In Alzheimer’s or Parkinson’s disease, a brain
spectrum — visible light. The remaining types of energy, PET scan that uses FDG as the tracer may detect
from low frequency radio waves to high frequency lower than normal FDG consumption in certain brain

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areas, meaning there is a reduction of metabolic Imaging (Phase I), Merck (Kenilworth, NJ; Phase I), Roche
activity in those regions — a possible indication of the (Basel, Switzerland; Phase I), and Janssen (Raritan, NJ;
early neurodegeneration. preclinical). Challenges that must be overcome when
Although FDG-based PET scans have emerged as a developing new radiopharmaceuticals for targets inside
useful tool in neurodegenerative disease, they are of the brain include developing an agent that will cross
not capable of clearly differentiating one type of the blood-brain barrier and be extremely target-specific.
neurodegeneration from another. To do that, disease-
specific radiopharmaceutical detection agents have
DETECTING CANCER
been developed. Tumor cells consume glucose at a much higher rate
(approximately 200 times higher) than healthy cells.
DaTSCAN (GE Healthcare; Chicago, IL) is a
This difference makes FDG-based PET scans a reliable
radiopharmaceutical that binds to cells in the brain that
method for detecting and monitoring cancer as follows:
release dopamine, a chemical messenger. Loss of this
type of brain cell is associated with Parkinson’s disease • After the body is injected, tumor cells consume the
(PD). PET imaging with DaTSCAN enables physicians to radiolabeled glucose (FDG) much more rapidly than
visualize the loss of dopamine-associated neurons, and non-tumor cells.
to diagnose PD. • The PET machine detects the gamma rays emitted
Preclinical efforts are now being directed at the by FDG.
development of radiopharmaceuticals for the detection • Areas of high activity are referred to as “hotspots”
of alpha-synuclein, a protein that is elevated in the brains and may indicate the presence of a tumor.
of PD patients. Alpha-synuclein elevation is thought to
PET scans may be used as part of an initial diagnosis,
occur earlier in PD progression than dopamine signaling
and as a way to monitor treatment. Tumors shrink in
irregularities; thus, the ability to detect it could enable
response to therapy while the spreading of cancer to
earlier diagnosis and intervention. This is considered
other parts of the body can point to metastases.
such an important goal in the field that in 2016, the
Michael J. Fox Foundation announced a $2 million
NOT SO RADIOACTIVE
dollar award to the first team that successfully develops
a PET tracer for the visualization of α-synuclein. Leaders The prospect of being injected with a radioactively-
in this race include AC Immune (Lausanne, Switzerland) labeled imaging agent may give some patients pause.
and ICBI International (La Jolla, CA). However, the total radiation dose received by that
patient is equivalent to the exposure caused by two
Three radiopharmaceuticals — Vizamyl (GE Healthcare),
routine chest X-rays according to the National Institute
NeuraCeq (Piramal Imaging; Berlin, Germany) and
of Biomedical Imaging and Bioengineering. This is due
Amyvid (Eli Lilly; Indianapolis, IN) — have been approved
largely to the rapid decay of the radioactive label used
for the detection of beta-amyloid protein plaques in
for PET tracers. Within a few hours of administration,
the brain, which could indicate the presence of the
there is no detectable radioactivity in the patient’s
Alzheimer’s disease (AD). They are approved for clinical
body. Patients are encouraged to drink plenty of
use in PET scans, however they are not considered to be
fluids after a PET scan to aid in flushing any remaining
stand-alone diagnostics. In other words, a negative scan
radiopharmaceutical out of the body.
may be used to rule out AD, but a positive result is not
enough on its own to diagnose. The ability to see inside of a patient’s body, directly
measuring metabolic activity or anatomical structure, is
The second type of abnormal protein cluster found in
critical to detecting, monitoring, and treating diseases.
AD patients’ brains are tau proteins. PET tracers for
With new discoveries in the field of imaging we can hope
the detection of tau are in development by several
to catch signs of disease sooner rather than later.
companies, including AC Immune (Phase I), Piramal

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CIRCADIAN RHYTHM & DISEASE Life Science Training from Industry Experts

AND THE BEAT GOES ON period. This is why most people feel disoriented after
traveling through several time zones, even if they get an
Earlier this week, the 2017 Nobel Prize in Physiology
adequate amount of sleep. These tiny molecular clocks
or Medicine was awarded to three American scientists
are signaling that it is later (or earlier) than the local
( Jeffrey Hall and Michael Rosbash, of Brandeis
time—and a whole range of physiological functions from
University, and Michael Young, of Rockefeller
body temperature to heart rate, blood pressure, and
University) for their work in deciphering the molecular
alertness levels respond accordingly.
basis of circadian rhythm – the 24-hour cycle that
governs the inner workings of all life on Earth. Although Clock proteins are regulated by a “master clock” in the
the work that garnered the Prize was done over twenty brain. Technically referred to as the suprachiasmatic
years ago, its implications to human disease and new nucleus, this master clock consists of about 20,000 nerve
therapies are still being worked out today. In this issue of cells just above the region where the optic nerve crosses
the Weekly, we’ll take a look at this Nobel science. into the brain. This location, called the optic chiasm,
explains why many of these neurons are sensitive
CIRCADIAN RHYTHM GOVERNS to light.
MORE THAN 24 HOURS
ON THE MARKET: HETLIOZ
Overnight shift workers, students pulling an all-nighter
to cram for a final exam, and business people rushing People who are totally blind cannot receive the proper
between time zones all share one thing in common: light input to control their master clock. As a result,
significant disruption to their circadian rhythm. This many suffer from non-24-hour sleep-wake disorder. It
roughly 24-hour activity cycle responds primarily to light becomes virtually impossible to fall asleep at standard
and darkness and is found in most living organisms— times, which severely impacts their ability to function
people, plants, animals, and even some microbes. professionally and socially.

Disruption in the cycle can cause more serious In July 2014, the FDA approved Vanda
consequences on top of a few days of disorientation. Pharmaceuticals’(Washington, DC) Hetlioz for patients
Abnormal circadian rhythms are correlated with with non-24-hour sleep-wake disorder. Hetlioz is an
insomnia, diabetes, increased cardiovascular events, agonist; it binds directly to and activates the melatonin
some types of cancer, Parkinson’s disease, and more. receptor. Melatonin is a sleep-inducing hormone with
levels increasing at the onset of darkness, linking it to
In this issue, we’ll take a closer look at how circadian
the sleep-wake cycle. Since melatonin production is
rhythms are regulated, examine some links with various
dysregulated in totally blind people, Hetlioz aims to
disease states, and find out how drug discovery efforts
normalize sleep patterns. Although melatonin itself is
are taking this important phenomenon into account.
available in pill or liquid form over the counter, synthetic
melatonin receptor activators such as Hetlioz are
PROTEINS ON THE CLOCK designed to be more stable.
The circadian rhythm is regulated by a combination of
environmental and internal cues. Environmental cues are IN DEVELOPMENT: RESET YOUR PRESET
based on light and dark, while internal cues are simply
Aptly-named Reset Therapeutics (South San Francisco,
“biological clocks.” These biological clocks are groups of
CA) is focusing on circadian-rhythm disruptions as they
interacting proteins in cells throughout the body.
relate to a whole range of diseases including type 2
The most extensively characterized biological clocks are diabetes, Cushing’s syndrome (elevated cortisol levels),
appropriately dubbed Clock proteins. The Clock proteins high blood pressure, obesity, sleep apnea, cancer,
interact with each other and regulate levels of expression and inflammation. Scientists at Reset have identified
of other proteins throughout an approximately 24-hour potential circadian-modulating compounds and are

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testing them in animal models of Cushing’s syndrome In most healthy people, blood pressure is tied to
and type 2 diabetes. circadian rhythm, naturally dropping between 12:00
AM and 3:00 AM. In people with high blood pressure,
PRECLINICAL: SHUTTING DOWN however, this drop often doesn’t occur naturally. By
THE CANCER CLOCK taking blood pressure medication at night, patients can
A possible link between circadian rhythm dysfunction restore this normal decrease in pressure.
and cancer has caught the eye of drug development. One Dosage timing may also be important for certain types
of the functions of the Clock proteins is to set restrictions of cancer drugs. For example, PARP1 inhibitors work
on when cells can divide, so circadian disruptions may by shutting down a DNA repair enzyme in cancer
affect cancer-related cell division. Researchers at the cells. The shutdown results in so much DNA damage
University of California, Santa Cruz are zeroing in on to the cells that they initiate a cell-suicide program
a protein known as PASD1, which is expressed in many known as apoptosis—the cancer cells pretty much end
different cancer cell lines. PASD1 interacts with the Clock up killing themselves! Researchers at University of
proteins, essentially interfering with their function and North Carolina (Chapel Hill, NC) have discovered that
shutting them down. Early preclinical work suggests DNA repair enzymes are more active later in the day,
that the inhibition of PASD1 causes the Clock proteins to translating into PARP1 inhibitors possibly being more
reactivate—making PASD1 a possible drug target. effective if given in the morning; inhibiting the enzymes
should be easier when they are not at their peak
TIMING IS EVERYTHING activity level.
Taking medicine at the same time every day is a good Circadian rhythms govern more than the 24 hours of
way to make sure a dose isn’t forgotten. For certain your day, and uncovering various mechanisms at the
medications, however, the time of day the pill is cellular level of the sleep-wake cycle might just open up
swallowed may influence drug efficacy. new avenues to treat a whole array of diseases.

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DNA VACCINES EXPLAINED Life Science Training from Industry Experts

MORE ON THE POWERFUL, ELEGANT Sometimes a viral subunit or subunits form what’s called
SIMPLICITY OF VACCINES a virus-like particle (VLP)–a protein structure whose
shape closely mimics a virus with none of its genetic
Last week, we overviewed vaccine development
material. In these cases, the body’s immune system
and manufacture, focusing on those that use whole
responds very robustly.
pathogens to protect us from a disease. This week, we
examine subunit and polysaccharide vaccines, which use A subunit vaccine can also derive from toxins produced
different strategies to fight infection. We also take a brief by dangerous bacteria. For example, Clostridium tetani
look at the US Food and Drug Administration (FDA)’s (tetanus) secretes tetanospasmin, a neurotoxin that
vaccine approval process. causes severe muscle spasms, potentially leading to
death. The vaccine contains inactivated toxin, which
A PART IS SOMETIMES GREATER helps develop antibodies that prevent future illness.
THAN THE WHOLE Because subunit vaccines contain none of a pathogen’s
The structure of viral cells is much simpler than that of genetic code, they are generally very safe.
our own cells. Despite the damage they can do, viruses
consist only of one or more strands of DNA or RNA, CARBS+PROTEIN=IMMUNITY
encased in a protein shell such as in the viruses you (SOMETIMES)
see below. In general, bacterial infections tend to be more difficult
to protect against by vaccine than viral infections.
That’s because the surface of some bacteria is covered
in long chains of carbohydrate molecules. Called
polysaccharides, they mask the bacteria’s proteins. This
“cloaking device” means the body doesn’t recognize the
threat and mount an immune response. But molecular
biologists and other scientists have discovered that it’s
possible to link polysaccharides to a harmless protein,
thereby coaxing an immune response. These vaccines
are known as conjugated polysaccharide vaccines because
the carbohydrate is conjugated (connected) to a protein.
The Haemophilus influenzae type B (or Hib) and some
This simple structure means that sometimes only part, types of pneumococcal and meningococcal vaccines are
or subunit, of a virus is enough to stave off infection. made this way.
Different subunit vaccines use different bits of a
pathogen. Often, they consist of nothing more than GETTING DOWN TO THE ESSENTIALS
one of a virus’s surface protein. Subunit vaccines work The basics of immunization have been around over a
because our immune systems recognize and respond century—use a disease-causing microbe, or just a part,
readily to these surface proteins. against itself. However, the latest step in the evolution of
For subunit vaccines, drug manufacturers alter yeast, vaccines takes a different tack, delving more deeply into
bacteria, or Chinese hamster ovary (CHO) cells to the building blocks of life—DNA.
produce a protein by transferring the gene encoding the Instead of immunizing someone with a whole pathogen
virus into them. These host cells make the viral protein, or fragment, a DNA vaccine injects only a small bit of a
which the manufacturer then isolates and formulates virus’s genetic code. Drug companies nestle the code
into the vaccine. Subunit vaccines include those for in plasmids— small, circular DNA molecules within a
hepatitis B and human papillomavirus (HPV). pathogen. As you can see below, the “visiting” DNA

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prompts the host to produce the target viral protein and TESTING, TESTING
consequent immune response within their own cells, but
How does a new vaccine come to market? The FDA
without an infection.
requires companies test vaccines for safety and
effectiveness on human subjects. The process differs
somewhat from clinical drug trials. Researchers test a
new drug on sick volunteers to see if it makes them well;
researchers administer an investigational vaccine to
healthy volunteers to see if it prevents illness.
Clinical vaccine trials involve three distinct steps.
• Phase 1 is typically a small study in which healthy
volunteers get the vaccine. Doctors monitor them for
side effects. If no unacceptable reactions occur, the
vaccine advances to the next stage.
• In Phase 2, more subjects, typically hundreds, get the
The key challenge for DNA vaccines is getting patients’
vaccine. The same number of subjects participate in
cells to accept the introduced DNA. So far, the most
the study without receiving the vaccine. Scientists
effective technique seems to be electroporation–
refer to this second group as the control. Both
delivering short pulses of electrical current to the patient
groups are at the same level of risk for contracting
with the vaccine. The electricity creates temporary pores
the target disease. Researchers observe the
in a patient’s cell membranes, enabling the DNA to enter.
“vaccinatees” for two years or more to see if they
The FDA has yet to approve any DNA vaccines for human contract the disease at lower rates than those in the
use. The prospect of DNA vaccines, however, presents control group. Researchers also monitor immune
some important advantages, which includes producing response by measuring levels of anti-pathogen
a strong immune response and somewhat easier antibodies in the participants’ blood.
manufacturing. Producing large volumes of viral gene-
• For Phase 3, even more subjects—often thousands-
containing plasmids still means growing lots of bacteria
-at high risk for the disease receive the vaccine and
in which to reproduce the plasmids, but purifying and
are monitored as described in Phase 2, from three to
formulating these vaccines is more straightforward due
five years. This trial also includes a control group.
to the relative simplicity of DNA’s structure. In addition,
DNA vaccines don’t require refrigeration, extending their
COCKTAIL FODDER:
shelf life and transportation time.
VACCINATION ACTIVATION
Inovio Pharmaceuticals (San Diego, CA) currently has
Getting a shot won’t make you sick. Sometimes, people
DNA vaccines for hepatitis B and C, and the Ebola, HIV,
feel mild symptoms such as fatigue, headache, and low-
and Zika viruses in the early stages of clinical testing.
grade fever after receiving a shot. These are signs that
an immune response is being activated.

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NATURAL BORN CANCER KILLERS Life Science Training from Industry Experts

FURTHER DOWN THE CANCER

TREATMENT ROAD WITH CARS
This past August, to much fanfare, the FDA approved the
first chimeric antigen receptor (CAR) T-cell therapy for
blood cancer. Called Kymriah (Novartis), it promises to
revolutionize treatment by genetically altering a patient’s
own cells to fight cancer. Less than eight weeks later,
Kite Pharma, now a part of Gilead Sciences (Foster City,
CA), had its new CAR T-cell therapy, Yescarta approved
as well.
Meanwhile, biotech companies continue to push the
boundaries of immunotherapy by creating engineered Need reminding how CAR-T cells work? Check out our
immune cells. WEEKLY on the topic.

A CAR THAT DOES WHAT? NATURAL BORN KILLERS

But first, what exactly is a chimeric antigen receptor? Our immune system inherently includes NK (natural
CARs are manufactured proteins that molecular killer) cells. They are the body’s first responders. At the
biologists and others engineer to appear on the surface first sign of illness, NK cells attack the infection for two
of a white blood cell such as a killer T-cell. This new, reasons. First, pathogens lack surface proteins called
revved-up receptor then targets the white blood cell to MHC1 that the body identifies as normal. Second, the
attack cancer cells. presence of abnormal proteins tells the body that the
invader poses a threat.
A CAR consists of:
Many immunologists believe that prompt action by NK
• Targeting domain: This part of the CAR exists outside
cells helps eliminate cancer cells early on– before they
the white blood cell. It is composed of an antibody
grow into a serious problem. However, in the early stages
that recognizes and docks onto a specific cancer
of tumor development, there are often not enough red
surface protein.
flags – abnormal proteins on the cancer cell surface – to
• Activation domain: This component kicks into gear tag them as dangerous. Engineering NK cells to display
once the targeting domain locks onto the intended a CAR “trains” them to recognize and respond to tumor
cancer surface protein. In CAR-T cells, the activation cells. Once activated, the CAR-NKs behave much like
domain signals T-cells to do three things: 1) make killer T-cells, releasing cytokines that bolster the immune
copies of themselves; 2) release signaling molecules response to the cancer cells--killing the nasty cells by
called cytokines (proteins that prompt other white injecting even nastier toxins.
blood cells to attack the tumor); and 3) finally—the
really good bit-- kill cancer cells. HOMEGROWN ISN’T ALWAYS BEST
CAR-NK cells have two important advantages over
CAR-T cells: safety and accessibility. CAR-T cells must
come from the patient’s own T-cells to avoid triggering
graft-versus-host-disease (GVHD). This potentially
deadly illness occurs when the patient’s immune system
responds badly to foreign tissue. Donor NK cells, in
contrast, don’t appear to cause GVHD.

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Besides avoiding potentially life-threatening reactions fragments of the alien cell’s proteins or antigens are
in already very ill patients, medical professionals can displayed on the macrophage’s surface. These leftovers
obtain donor NK cells relatively easily—for example, help activate some of the immune system’s other
from umbilical cord blood. Labs modify these donor cells defenses, such as killer T-cells.
to express a CAR, which can then be given to the patient.
Removing and engineering a patient’s own T-cells, then MMM…CANCER?
transfusing them back into the patient is much more Researchers are now exploring the potential power of
time-consuming. The ability to more easily use donor NK CAR-macrophages to destroy specific cancer cells. The
cells means that biotech companies can create “off-the- enhanced macrophages will simultaneously activate
shelf” products for this type of CAR therapy more readily. other immune cells to also recognize and attack those
In addition, the resulting lower production costs mean same antigen-bearing cells. Like other macrophages,
more available treatments. CAR-macrophages can penetrate solid tumors much
more effectively than “plain old” T-cells.
TEST-DRIVING CAR-NKS
If a typical T-cell does make it into a solid tumor, the
This past June, scientists at the MD Anderson Cancer cancer’s own defenses makes short work of it. In
Center (Houston, TX) started a Phase I/II trial of CAR- contrast, by modifying macrophages to treat solid
NK cells. The research focuses on patients with chronic tumors, doctors may be able to effectively get at cells
lymphocytic leukemia (CLL), acute lymphocytic leukemia inside the tumor. At the same time, the super-powered
(ALL), or non-Hodgkin lymphoma. The trial cells macrophages will “wake up” the patient’s suppressed
contain a “suicide” gene that is triggered by excessive T-cells to fight the cancer as well.
inflammation. Researchers hope this built-in safety
Preclinical data from CARMA Therapeutics
feature will reduce problems caused by overactive
(Philadelphia, PA) shows that its scientists can modify
immune responses in patients from earlier trials of
CAR-macrophages to recognize and engulf different
CAR-T cells.
types of solid tumor cells. They’ve also demonstrated
Researchers elsewhere are also looking into the that infusing cancerous mice with tumor-specific CAR-
development of CAR-NK cells that treat other cancers— macrophages leads to long-term tumor control and
specifically targeting the HER2 protein in breast and longer survival. CARMA plans to begin clinical testing the
ovarian cancers. effect of CAR-macrophages on specific ovarian cancers
in 2019.
CHOWING DOWN ON DISEASE
As biotech companies continue to translate these new
Of course, NKs, modified or otherwise, aren’t the only
applications of CAR into treatments, both patients and
white blood cells going toe-to-toe with cancer. Another
doctors can look forward to seeing an increase in the
approach involves the immune system’s scavengers
number of different types of cancer that respond to
or macrophages. “Macrophage” comes from Greek,
these cutting-edge immunotherapies.
meaning “big eater.” These cells kill invading or diseased
cells by surrounding and digesting them. Leftover

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OFF-COLOR: THE SCIENCE BEHIND
COLOR VISION DEFICIENCY Life Science Training from Industry Experts

You’re at the supermarket, puzzling over whether those Women have two X chromosomes; men only one. With
peaches for the pie are ripe. Maybe you’re watching your two copies of each opsin gene, it’s unlikely both would
child’s soccer team, and struggling to separate the Green contain mutations interfering with color perception.
Hornets from the Scarlet Knights. As if determining By far, most cases of genetic color blindness come
offsides isn’t hard enough! Or more seriously, you’re from genetic mutations that code for red or green-
approaching a stoplight on a busy street and can’t tell detecting opsins. Consequently, those affected have
if the signal is red or green. People with color vision trouble distinguishing shades of red or green. Rather
deficiency (CVD), more commonly known as color than responding to only clearly distinct wavelengths
blindness, face challenges like these all the time. That’s of light, the mutated opsin detects a broader range,
not to mention the inability to fully experience some of essentially bleeding over into the neighboring detection
the world’s wonders—the blazing of sugar maples in the wavelength. The brain detects color by comparing
fall or the first flush of April’s flowers. signals from two different types of cone cells. Too much
overlap between signals interferes with a person’s
TERM OF THE WEEK: perception of color.
PHOTORECEPTOR CELL
While most people with color vision deficiency have
Photoreceptors are cells in the eye’s retina that respond
trouble distinguishing colors in the red and green
to light and convert its signals into information the brain
parts of the spectrum, other conditions exist, such as
uses to create visual representations of images. We have
tritanomaly—reduced sensitivity to blue light.
two kinds of photoreceptors: rods and cones. Rods work
in low levels of light and detect only black and white. Much more rarely, two different color-detecting opsins
Cones enable us to see colors. are affected, which results in significantly reduced
color vision. Even though people with this condition
Photoreceptors contain light-sensitive proteins called
retain one-color-detecting opsin, the brain needs to
opsins. Opsins change chemically in response to light.
compare signals from different types of cones to see
Photoreceptors detect those changes, enabling them
color. The most severe and rare form of CVD is called
to convey visual information. Rods contain only one of
achromatopsia—true color blindness. People with this
these proteins, called rhodopsin. Cones, in contrast,
condition perceive no color at all, and suffer from other
contain three different opsins, one of which responds to
vision problems as well.
red, green, or blue wavelengths of light. For most people,
our opsins bring us the world in technicolor. IMPROVING THE OPTICS
SEEING RED—OR NOT There is no cure for color blindness at present.
However, two remarkable adaptive technologies exist.
The most common types of color blindness are inherited.
A company called Enchroma (Berkeley, CA) produces
In these cases, the genes for one or more cone opsin
glasses that help people with red-green color blindness
are defective and make abnormal proteins. Physical or
perceive colors normally. They combine the latest in
chemical damage to the eye, optic nerve, or brain can
lens development with color perception neuroscience.
also cause color blindness.
Enchroma’s proprietary technology filters specific
wavelengths to create sharp distinctions between red
YES HONEY, THE DRESS IS PURPLE and green, significantly improving color vision. Available
Color blindness affects far more men than women. off-the-shelf online or at vision centers nationwide, the
The National Eye Institute (Bethesda, MD) estimates glasses can incorporate prescription lenses.
that nearly one in 12 men are color blind, versus one in
Colormax (Timonium, MD) also markets color vision
200 women. This difference arises because genes for
correction contact lenses and glasses that filter specific
the different opsin proteins are on the X chromosome.
wavelengths of light. The lenses are custom-made

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and so require a thorough in-person exam. Colormax PHILOSOPHICAL QUESTION:
guarantees that the lenses will enable wearers to pass RED OR RED?
the color vision tests required for certain occupations,
How does a person know if the red they see is the same
such as firefighter, pilot, and railroad engineer.
red someone else sees? Even among the majority of
TINKERING WITH THE FAULTY GENES people, who see color “normally,” there are probably
variations in color perception. That’s because the eye
Scientists at Adverum Biotechnologies (Menlo Park, CA)
and brain work in concert to translate light into color.
are looking at biotech approaches to treating inherited
Add to that the physical differences in saturation,
forms of color blindness. Possible gene therapies could
hue, and tone and the panoply of color names in
entail delivering a correct version of the mutated gene.
the English language alone—crimson, blood, garnet,
Adverum’s work stems from gene therapy treatment of
vermilion, maroon…and we’ll all be seeing red for a
CVD squirrel monkeys at the University of Washington in
loooonnng time.
Seattle, WA. .

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THE SCIENCE BEHIND OPIOID ADDICTION Life Science Training from Industry Experts

THE SCIENCE BEHIND OPIOIDS Opioid pain medications, such as Purdue Pharma’s
(Stamford, CT) Oxycontin and Endo Pharmaceuticals’
Concerns over the opioid epidemic continue to grow,
(Malvern, PA) Percocet, work by binding to mu receptor
with deaths from narcotic overdoses the leading cause
proteins on the surface of cells in the central nervous
of death in people under 50 last year. Nearly half of
system (CNS) —think brain and spinal cord. While the
those deaths were attributable to prescription opioids.
CNS is tasked with relaying pain signals, opioids decrease
The directors of both the Center for Disease Control
the excitability of nerve cells delivering the message,
(Atlanta, GA) and the Food and Drug Administration
resulting in pain relief—along with a feeling of euphoria
(Silver Spring, MD) have called for urgent action on the
in some users.
crisis, and President Trump declared the opioid epidemic
a public health emergency. LESSENING THE PAIN
This stark reality highlights the dark side of a class of Short term medical used of opioid painkillers rarely
treatments serving a vital need. Opioid pain medications leads to addiction when properly managed. Due to
manage the severe short-term or chronic pain of millions the euphoria-inducing effects of the drugs, long term
of Americans. While these medications mitigate needless regular use, or use in the absence of pain, may lead
suffering, the US government, healthcare industry, and to physical dependence and addiction. And because
medical community are joining forces to battle against regular use increases drug tolerance, higher doses are
opioid abuse and addiction. required to achieve the same effect, leading abusers to
Here at the WEEKLY, we always wonder: What is the consume pain pills in unsafe ways such as crushing and
science behind the headlines? So, let’s talk about how snorting or injecting the pills. According to the Centers
pain medications work, the different types on the for Disease Control, 91 Americans die every day due to
market, and the approaches to developing less addictive opioid overdose, which includes prescription opioids and
versions of opioid drugs. heroin. At the same time, chronic pain is also a serious
problem, affecting approximately 100 million U.S. adults,
OPIOIDS VS. NSAIDS while millions of others suffer acute pain due to injury
There are two main categories of pain medications, or surgery. The medical need for these drugs is very real
opioids and non-steroidal anti-inflammatory drugs despite the dark side.
(NSAIDs). Although these two categories of drugs work The answer to developing less addictive drugs may
differently, they do share one thing in common: both be found in a drug that blocks pain without inducing
are derivatives of natural products. NSAID commonly euphoria. These new drugs will need a different
known as Aspirin was developed by Bayer (Leverkusen, mechanism of action than traditional opioid drugs, which
Germany). It is a synthetic version of an extract from bind to the mu receptors of cells inside the CNS. Cara
willow tree bark. Opioids are synthetic versions of opium Therapeutics (Shelton, CT) is developing drugs that
and morphine, which come from poppy flowers. bind to a different type of opioid receptor, the kappa
Aspirin works by stopping an enzyme called opioid receptor. These receptors are present on sensory
cyclooxyrgenase 1 (COX-1). Once stopped, COX-1 can nerves outside of the CNS. Preclinical studies suggest
no longer produce prostaglandins and thromboxanes. that targeting these receptors could be effective at
Prostaglandins and thromboxanes are produced in reducing pain without driving addictive behaviors. Their
response to injury or infection and cause inflammation, lead candidate, CR845, is currently in Phase III clinical
which is associated with symptoms of fever, swelling, and testing for post-operative pain and pruritus (severe
pain. Other NSAIDs, such as ibuprofen and naproxen, itching), and in Phase II clinical testing for chronic pain.
also work by inhibiting COX-1 or its sister enzyme COX-2. Cara Therapeutics is also developing compounds that
selectively activate cannabinoid (CB) receptors outside
of the CNS. It’s interesting to note that CB receptors

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inside the CNS are linked to marijuana’s psychoactive to and activate the mu-opioid receptors, resulting in pain
qualities. Cara’s lead CB receptor activator, CR701, is in relief and mild euphoria without the detrimental side
preclinical development. effects associated with opioid drugs such depressed
Hydra Biosciences (Cambridge, MA) is developing small respiration, motor impairment, and addiction. Scientist
molecule drugs that inhibit ion channels, proteins on have tried before to develop safer pain medications
the surface of nerve cells that help transmit pain signals. based on endogenous opioids, but have not been
This plays a critical role in sending the pain signal to successful due to the molecule’s instability. The Tulane
the brain, yet because it works on nerves outside of team created a stable derivative of endomorphin that
the brain, it has less of a potential for addiction. Hydra binds to the mu receptor in such a way that pain relief
is currently in Phase II clinical studies of HX-100 for the occurs, but not the negative side effects listed above.
treatment of painful diabetic neuropathy. Clinical testing is expected to begin within a few years.

Centrexion Therapeutics (Baltimore, MD) lead ANTIDOTE TO AN OVERDOSE

candidate is a derivative of capsaicin, a naturally-
Overdosing can be fatal since respiratory failure
occurring compound found in chili peppers. Capsaicin
occurs at high opioid concentrations. If an overdose is
has pain relieving properties and is used as a natural
suspected, the individual should be treated as quickly
remedy. Centrexion’s lead candidate, CNTX-4975, is a
as possible with naloxone—a “competitive antagonist”
highly potent, synthetic form of capsaicin designed to be
of the mu-opioid receptor. Simply put, a competitive
administered via injection into the site of pain. CNTX-
antagonist binds the receptor without activating it.
4975 targets the capsaicin receptor, an ion channel
Since naloxone doesn’t activate the receptor, it doesn’t
protein on the surface of nerve cells. When CNTX-4975
have any pain relieving or euphoria inducing qualities;
binds the capsaicin receptor, the influx of calcium ions
rather, it prevents the opioid drugs from binding. It may
results in desensitization of the nerves, making them
also displace opioids that have already bound the mu
unresponsive to other pain signals. This effect can
receptor, aiding in the stoppage of an overdose.
last for months, and only affects nerves near the site
of injection. Centrexion recently announced positive
Phase IIB results for CNTX-4975 in a study testing its
COCKTAIL FODDER: RUNNER’S HIGH
effectiveness against pain in knee osteoarthritis. The Some folks love to run; others avoid it at all costs.
drug is also in Phase II clinical studies for Morton’s This might be explained by inherent differences in
neuroma, a sharp pain in the foot and toe caused by a sensitivity to the natural opioids called endorphins
thickening of the tissue around one of the nerves leading that are released during exercise. Not everyone
to the toes. experiences the “runner’s high”— feelings of calm and
mild euphoria – in much the same way that not everyone
Earlier this year, Nektar Therapeutics (San Francisco,
experiences euphoric feelings from pain medications.
CA) announced positive Phase III results of their
These differences may help to explain why some people
addiction-resistant opioid, NKTR-181. Like traditional
enjoy exercise and others don’t, and why some people
opioids, NKTR-181 works by entering the brain and
get addicted to opioids—while others can take them or
activating mu-opioid receptors. However, the molecule is
leave them.
designed to enter the brain much slower than other mu-
receptor activators, reducing the feelings of euphoria With several of the opioid alternatives outlined here
while offering pain relief. already showing success in clinical studies, there is hope
on the horizon for patients who suffer from chronic pain
Researchers at Tulane University (New Orleans, LA)
but want to avoid the risk of addiction. Next week, we’ll
have developed a compound that is derived from the
explore the use of medical devices in pain management.
endogenous opioid, endomorphin. Endogenous opioids
are chemicals produced naturally by the body that bind

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BYE-BYE OPIOIDS? INTRODUCING
ELECTROCEUTICALS Life Science Training from Industry Experts

MEDICINE-FREE PAIN MANAGEMENT

Migraine relief without drugs? No “digestive issues” due
to pain meds after surgery? Better still, no worry about
addiction after that appendectomy or hip replacement?
Sounds a bit science-fictiony, does it not?
The news reminds us nearly every day of the profound
need for pain management without opioids. As you read
last week, alternatives to analgesics such as morphine,
codeine, or fentanyl exist. But science also takes us
beyond medicine: researchers have developed devices
that control pain and inflammation electronically. This
drug-free approach is called electrostimulation–using Groups of neurons form nerves outside the central
low levels of electricity to zap pain. nervous system. Nerves transmit sensory information
such as touch, temperature, and pain. Different kinds
THE BODY ELECTRIC of nerves respond according to the specific receptor on
their surface. Pain receptors detect chemicals that tissue
How does one feel pain in the first place? Pain, and more
damage releases. The pain signal then zooms along the
pleasant sensations (the taste of chocolate, for instance)
nerve to the central nervous system for interpretation
come courtesy of the nervous system. This system
and response by the brain. Pain can also be caused
serves as the wiring that enables the machines of our
by direct damage to the nerves themselves (think
bodies to move, experience, and interpret the world.
pinched nerve).
Our nervous system has two main parts that work
in tandem: the central and peripheral nervous SLAMMING THE GATE ON PAIN
systems. The central nervous system functions as the
As you no doubt noticed, pain perception is complex.
“switchboard” that sends and receives information from
Remember the last time you bashed your knee? Did you
peripheral nervous system’s local lines. The peripheral
rub it? The Gate Theory of Pain says this rubbing action
nervous system is a vast network of nerves found
does temporarily reduce pain. In that moment, signals
everywhere in body.
from your non-pain nerves block signals from your pain
The central nervous system is made up of neurons. nerves. In other words, non-pain nerves prevent pain
These cells send and receive signals electrochemically, from reaching your brain.
through signaling molecules called neurotransmitters.
Electrostimulation relies on the body’s ability to run
These chemicals are converted into electrical signals
interference on pain. It uses electricity to treat chronic
in the neuron. Here’s how: when a neurotransmitter
pain by stimulating a patient’s peripheral nerves or
reaches one edge of a neuron, the dendrite, it opens
spinal cord. In either case, a small pulse generator sends
ion channels in the cell membrane. These tiny openings
electrical pulses to the nerves or spinal cord, “shutting
allow the positively charged sodium ions to pass
the gate” on pain.
through. These positive ions travel down the other side
of the neuron through an extension called an axon. Several companies already have electrostimulation-
When the electrical charge reaches the end of the based devices on the market, including:
axon, it signals the release of other neurotransmitters, • Medtronic (Fridley, MN). Its Intellis spinal cord
which then switch on other neurons and the chain of neurostimulation system gained FDA approval
sensation unfurls. in September and European Union approval for
managing chronic pain earlier this month. This

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 small, implantable neurostimulator communicates Federal agencies, large pharmaceutical companies,
wirelessly with a Samsung tablet that a doctor uses and small start-ups are taking note. The National
to adjust pain relief and monitor patients’ activity. Institutes of Health (Bethesda MD) has established
More activity indicates that the patient’s level of pain the Stimulating Peripheral Activity to Relieve Conditions
is lessening. (SPARC) program to fund the basic exploration of how
• Stimwave (Pompano Beach, FL). This company’s peripheral nerves’ electrical signals control the function
latest device, the StimQ Peripheral Nerve Stimulator, of internal organs. This effort may lay the groundwork
was approved in August for chronic pain. Tiny for electroceuticals–the next generation of tiny
enough to be inserted by needle, it communicates neurostimulation devices.
wirelessly with an external transmitter that In the private sphere, GlaxoSmithKline (GSK; Middlesex,
delivers electrical pulses. The device is also safe U.K.) leads the charge. The corporation is heavily
to wear in MRI machines. This innovation makes involved in creating a Nerve Atlas to map the nervous
neurostimulation available to people whose system’s role in disease. GSK is also investing in start-
treatment requires ongoing imaging. ups such as SetPoint Medical (Santa Clarita, CA),
• Abbott (North Chicago, IL) also offers which has already produced clinical data supporting
neurostimulation systems. Their most recent is the the use of electroceuticals on rheumatoid arthritis .
BurstDR system. Developed by St. Jude Medical in St. Another of GSK’s notable investments in electroceuticals
Paul, MN, this proprietary technology delivers bursts was the launch of Galvani (Stevenage, England) in
of electrical stimulation in a way that mimics the 2016, in partnership with Verily (San Francisco, CA).
body’s natural response to pain. Galvani is developing electroceuticals for chronic
conditions including Type 2 diabetes, autoimmune, and
THE LATEST BUZZ: NEW endocrine disorders.
APPLICATIONS AND NEW TECH
COCKTAIL FODDER: THE DOWNSIDE
While neurostimulation can treat chronic pain, the
OF FEELING NO PAIN
technology may provide other therapies as well.
Because nerve signals permeate the entire body, Feeling no pain sounds great, doesn’t it? Yet people born
modulating them may work for other medical with congenital analgesia suffer greatly from injury and
conditions. For example, neural signaling partially illness, because they never feel the pain associated with
controls inflammation, the body’s response to injury. them. In a BBC interview, sufferer Steven Pete talked
This suggests that neurostimulation may treat about breaking a limb roller skating as a boy, noticing it
inflammatory disorders such as Crohn’s disease and only when friends pointed out the bloody wound caused
rheumatoid arthritis. by the bone protruding from his leg! Fortunately, this
inherited nervous system disorder is extremely rare,
with only 20 or so cases known in the medical literature.

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ZINC FINGER NUCLEASES Life Science Training from Industry Experts

CATCHING THE RIGHT BREAK genome and the nuclease cuts the specific location. The
double-stranded break (DSB) or cut activates the non-
How are ZFNs made? To start, zinc finger proteins (ZFP)
homologous end joining (NHEJ) repair pathway, this most
are sequence-specific, DNA-binding proteins that
often results in a disruption of a gene, useful for gene
activate gene expression. They are engineered to
“knockouts”. If a repair template is delivered at the same
recognize unique sites within the genome. While ZFPs do
time as the break, the homologous directed repair (HDR)
not have the ability to cut DNA on their own, scientists
pathway kicks in. This method is useful to “knockin” a
can fuse a ZFP together with a DNA-cutting enzyme
gene. This video explains a potential application for zinc
called nuclease—the “N” in ZFN. The marriage of ZFP to
finger nucleases in HIV.
nuclease creates ZFN.
How do ZFNs work? Zinc finger proteins bring the
zinc finger nuclease to the engineered location of the

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PLANTS THAT HEAL Life Science Training from Industry Experts

NATURE’S MEDICINE CABINET history in western medicine as well. It’s the active
ingredient in many cold medicines, including Sudafed.
Where does medicine come from? Before it gets to
Doctors also prescribe ephedrine to prevent low blood
your medicine chest? Before you purchase it from
pressure during spinal anesthesia.
your neighborhood drugstore? Next time you’re hiking
through a forest or gazing at your pretty screensaver Indian snakeroot (Rauwouofia serpentina) is also
of the Olympic Peninsula, think of this: the magic that featured prominently in early Chinese and Indian
relieves a throbbing headache or lowers your dad’s medicine. It contains an indole alkaloid, reserpine.
blood pressure may well have started with a plant. Studying this chemical helped neuroscientists
learn how neurotransmitters such as serotonin or
Historically, humans have looked to the world around us
dopamine govern depression. Reserpine is prescribed
for what we need. Plants in particular offer the power to
for hypertension and psychosis. Mahatma Gandhi
feed, heal or potentially harm us. They produce an array
reportedly relied on reserpine to calm his nerves.
of bioactive compounds to survive and thrive. Some of
these substances are also biologically active in people FROM FOREST TO PHARMACY
too. Sometimes, years of hard work and testing turn one
Plants beyond those used in traditional medicine
of these compounds into drug we can use.
continue to influence modern medicine. In fact, for the
DOCTOR MOM IN THE past 25 years, approximately one quarter of prescribed
FOREST PRIMEVAL medicines in the US contain plant derivatives.

Imagine early humans, scrabbling for survival out on According to some historians, the modern
the savanna somewhere. One of the toddlers develops a pharmaceutical industry began as early as the mid-
burning fever. Desperate, the mother remembers those 1800s. Others say the early 1900s. Regardless, the first
berries along that new stretch of riverbank. She asks commercial drugs derived largely from plant extracts.
herself, “What if?” Let’s say the berries cure the baby’s They include these old favorites:
malady. Voila—the first medicine, courtesy of Dr. Mom. • Digitalis. Cardiac glycosides from the foxglove plant
stimulate cardiac muscles and are used to treat
THE DIVINE FARMER congestive heart failure. Careful! Every part of these
As long as humans have eaten plants, we’ve funky, beautiful flowers is poisonous to dogs, cats
experimented with their health effects. The Chinese and humans.
were likely the first to formalize their knowledge into • Aspirin. Aka acetylsalicylic acid. Salicin, the bioactive
what we call Traditional Chinese Medicine (TCM). ingredient behind this most basic medicine comes
According to Chinese lore, the mythical emperor from willow tree bark. Many of us use it to manage
Shennong, sometimes known as the Divine Farmer, pain, fevers and inflammation. My mom uses it to
commanded his herbalists to record the effects of help prevent stroke.
medicinal plants approximately 5,000 years ago.
• Quinine. This alkaloid is a product of bark from
Ethnobotanists and others have examined many TCM the native South American cinchona tree. In the
plants for use in pharmaceuticals. Some have yielded past, quinine was famously used against malaria.
ingredients regularly used in western medicine. It is currently used to treat patients with lupus or
They include: rheumatoid arthritis. For malaria, a derivative of
Mahuang or Ephedra (Ephedra sinica). Mahuang was another plant ousted the cinchona—artemisinin,
used to relieve asthma, or at least wheezing and which comes from sweet wormwood.
congestion. Scientists now know that mahuang contains Through the 1930s, pharmacological research remained
an alkaloid called ephedrine. Ephedrine has a long focused on screening natural products. Plants, bacteria

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and fungi were tested for their potential anti-cancer, rule is that these products are considered safe until
anti-inflammatory or anti-bacterial properties. proven unsafe.
In 1955, the National Cancer Institute began screening Some supplements are whole plant extracts, containing
natural products for cancer treatment. This program multiple bioactive ingredients. If a pharmaceutical
yielded a potent anti-cancer compound, taxol. Taxol company wants to develop a drug from a supplement,
came from the bark of the pacific yew, an evergreen it generally needs to isolate and test a single compound
native to the Pacific Northwest. In 1993, the FDA from the whole plant extract. Companies such as
approved a modified version of taxol, Paclitaxel, for Sanofi (Paris, France), Novartis (Basel, Switzerland) and
chemotherapy. Today, Paclitaxel is on the World Health Pfizer (New York, NY) are investigating plant-derived
Organization’s List of Essential Medicines. compounds in their drug discovery pipeline. One smaller
Scientists these days often synthesize new drugs using company, Napo Pharmaceuticals (San Francisco, CA),
organic chemistry or recombinant DNA techniques. isolated a compound from the tropical croton plant
These medicines specifically target areas in the (Croton lechery). It won FDA approval as an anti-diarrheal
human body where diseases originate. Many in the for HIV patients in 2013.
pharmaceutical and medical communities no longer Here are a couple plants and one fungus currently
consider natural products a primary source of new marketed as supplements. Remember, caveat emptor—
therapeutic compounds. guinea pigs are us.
However, the plant kingdom remains a biological White mulberry. Dried fruit from the white mulberry
treasure trove. A 2016 study by England’s Royal Botanic tree tastes a bit like raisins. The fruit contains fiber,
Gardens Kew estimates our amazing planet supports protein and vitamins and deoxynojirimycin, or DNJ.
about 390,900 species of plants. So far, researchers have DNJ is a glucosidase inhibitor that acts upon the sugar
screened only about six percent for biologic activity in metabolism pathway to prevent a spike of blood-sugar
humans. Who knows what pharmaceutical miracles may levels after eating. DNJ may prove useful for diabetics.
originate in this immense diversity? Devil’s claw. This small desert plant is native to
Southern Africa. In the early 1800s, Europeans
A SUPPLEMENT IS NOT A DRUG introduced this plant at home, where its root became
More and more, people use plant-based supplements a popular remedy for pain and osteoarthritis in France
like echinacea to combat their colds or valerian to and Germany. Harpagide, a glycoside from devil’s
help them sleep. Scientist are investigating some claw, appears to control inflammation. Devil’s claw has
supplements for their medical potential. However, also gained some traction with dog and horse owners
it’s vital to remember that the FDA does not consider because those animals also experience relief after taking
supplements drugs. So, what are the differences the herb.
between a supplement and a drug?
Chaga. Fungal tea, anyone? Strange but true—look
The FDA evaluates and monitors the development of for a chaga concoction on the chalkboard of your local
prescription medicine in the US. The agency defines coffee shop soon. This fungus grows on hardwood
drugs as substances intended to diagnose, treat or trees in chilly, northern forests (Maine, Alaska, Canada,
prevent disease. They must pass clinical trials before Siberia). Packed with antioxidant enzymes, phytosterols
being approved for consumer use. The process often and other bioactive compounds, preliminary research
takes twelve years or more and incredible resources— suggests the ability of chaga extract to reduce
intellectual and monetary. inflammation, reduce fatigue and boost immunity. No
In contrast, the FDA has no power to regulate verdict on taste though!
supplements. Under the Dietary Supplement Health Next week, we’ll move from the forest and the garden
and Education Act, the FDA must treat supplements like to the lab—explaining key steps researchers take to
food. The law doesn’t require supplements be tested for identify and move potential medicinal compounds into
safety, purity or quality. They include minerals, vitamins, clinical trials.
amino acids and other biological substances. The general

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DRUG DISCOVERY 101 Life Science Training from Industry Experts

ON THE ROAD TO NEW MEDICINES X MARKS THE MOLECULE

For most of the 20th century, we discovered new drugs Researchers identify their molecular targets by
by trial and error. Scientists investigated countless answering one incredibly complex question: How does
unrelated compounds in animals to see which improved diseased tissue differ from healthy tissue? Scientists look
disease symptoms. For instance, in the 1950s and 60s, at two biological clues: differences in gene sequences
British scientists at Boots Laboratories tested hundreds (mutations), and changes in the levels of specific
of unrelated chemicals on guinea pigs searching for an proteins. For example, does a lung tumor biopsy show
alternative to aspirin for treating pain and inflammation. mutations absent in healthy lungs? Are levels of protein
This scattershot approach consumed vast amounts of expression higher or lower in the diseased tissue? If
time and resources, with limited success. the answer to either question is yes, scientists have a
potential target for their ‘as yet to be developed’ drug.
PICKING UP THE PACE—
RATIONAL DRUG DISCOVERY
Beginning in the 1980s, scientists began to take a new
tack in developing drugs. They adopted an approach
known as rational drug discovery or mechanism-based
drug design. In this method, researchers first seek to
understand a disease at the cellular level, and ideally,
identify its mechanism. Once the cellular mechanism
becomes clear, scientists can identify a drug target:
the molecule (usually a protein) involved in the illness
that the ‘as yet undeveloped drug’ will hopefully act on.
Although still labor and resource intensive, rational drug
discovery tends to produce more highly-effective drug
TESTING, TESTING
therapies than the old trial and error approach.
In the quest for new medicines, as in the rest of life, few
PROBLEMS WITH PROTEINS things are as simple as we hope. Just because a protein
plays a fundamental role in an illness doesn’t always
Identifying a target molecule for a drug to act upon
make it a good drug target.
requires years of basic research on a disease’s biology.
Most diseases stem from issues related to how our Consider the following scenario: Researchers discover
bodies produce proteins. Significant underproduction a specific protein target that affects the course of a
of a protein causes disease; for instance, little to no disease—great! They then design a drug to inhibit
insulin production leads to Type 1 diabetes. In contrast, said target—also great! Problem solved, right? No so
overproduction of a protein can also produce disease; fast. Sometimes a different protein can quickly take
too many of the growth factor receptor HER2 can result over the target’s function, nullifying the new drug’s
in certain breast cancers. Finally, errors, or mutations, hoped-for effect. It’s back to the drawing board for the
sometimes affect protein function. This is the case when disappointed scientists.
a mutated tumor suppressor protein no longer controls In other cases, inhibiting a specific target achieves a
cell division and cancer develops. Understanding which goal – say halting cancer cell growth – but results in
proteins correspond to a particular disease means unexpected side effects, such as irregular heartbeat.
researchers can design drugs to go after that disease- It’s crucial that scientists validate that when a target
causing protein specifically. is altered, that change to the target is both safe
and effective.

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Researchers verify the suitability of target molecules by TERM OF THE WEEK: BIOMARKERS
answering two important questions: does the target play
One important tool in drug discovery and development
a key role in the disease? and will targeting the molecule
are biomarkers. Biomarkers are molecules that doctors
likely be safe and effective? Validation most often
and others use to measure normal or abnormal
includes cell-based (in vitro) and animal (in vivo) testing.
biological processes in the body. They can help show the
The goal of many drug therapies is to stop or slow a
effects of a disease or therapeutic intervention.
target’s activity. Consequently, many tests, or validation
assays, seek to find out what happens when a drug puts LDL cholesterol is a classic biomarker familiar to
the brakes on a molecular bad actor. anyone over the age of 35. Numerous studies have
linked it to heart disease. LDL cholesterol makes an
One of the most popular testing methods is RNA
ideal biomarker because of these three reasons: it
interference (RNAi). This technique can rapidly block
circulates in our blood and can be obtained by a simple
production of a particular protein. It does so by
blood draw; it can indicate heart disease early on, since
destroying the messenger RNA that codes for the
someone can have elevated cholesterol long before
protein. This destruction can quickly demonstrate what
developing atherosclerosis; and it changes as the
happens to a cell when protein production ceases, thus
disease progresses. In developing drugs, researchers
mimicking the effects of a strong inhibitor drug.
have used changes in LDL as a surrogate endpoint.
If a cell model shows promise, researchers move on Surrogate endpoints are outcomes that researchers can
to an animal model. Often, scientists use “knockout” measure to determine relatively quickly whether the trial
mice – in which they have disrupted a target gene that intervention benefits the participants.
codes for the target protein. Researchers pose similar
In the case of heart disease, researchers establish
questions to those asked in cell-based testing, but on
a target cholesterol level in lieu of waiting years or
the larger scale of the whole animal. Do the mice get
even decades to demonstrate a change in disease
cancer or Parkinson’s disease or heart disease when the
progression. In many cases, measuring biomarkers has
target gene is tampered? Because in vivo testing allows
turned out to be key to running shorter, less expensive
researchers to examine whole-body effects, it provides
clinical trials. We know now that statins, which lower
valuable information about target safety that in vitro
cholesterol, can increase longevity because patients
testing simply cannot.
have taken them for more than 30 years. Initial trials,
however, simply showed a decrease in patients’
blood cholesterol.
Identifying and validating drug targets are only the first
steps on the long road to safe, effective new medicines.
Tune in next week to find out how researchers identify
therapeutics that might make it into clinical trials.

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DRUG DISCOVERY 201 Life Science Training from Industry Experts

WE WANNA NEW DRUG GLOW

“One that won’t make me sick/ One that won’t make me Imagine “Enzyme Z” causes “Disease Z.” Its cure lies in
crash my car/ and make me feel three feet thick…” Huey shutting down, inhibiting “Enzyme Z.” Enzymes feature
Lewis is singing about love, but he voices very human active sites, a kind of pocket that binds various small
concerns when it comes to the medicines that heal molecules. Researchers manipulate these small molecule
bodies and minds. inhibitors to light up--emit a fluorescent signal if they
bind (shut down) the enzyme.
Last time, the Weekly explored how researchers identify
drug targets—the molecules in our bodies that make
good candidates for therapeutic action. It’s a painstaking
but strategic process, and only the first step toward
Huey’s mythical remedy.

THERAPEUTIC CHOICES
There are small molecule drugs and large molecule
drugs. Small molecule drugs include medicines most
of us know, such as aspirin and over-the-counter cold
remedies. In contrast, large molecule drugs, also known
as biologics, are often specialty drugs, delivered through
injection by a healthcare professional.
NO GLOW
Therapeutic choice hinges on a disease’s origins. If an
Scientists use similar colorimetric tests for large
illness originates outside the cell, a biologic is a logical
molecule drugs such as antibodies. Let’s say that
choice since it is too big to enter the cell. Though more
activated “Receptor X” turns on a particular gene that
expensive to produce, biologics typically cause fewer
in turn causes cancer. Scientists can engineer cells so
adverse reactions than their smaller counterparts.
when Receptor X is activated, the cell produces a green
Small molecule drugs attack disease from within the fluorescence and when it is not activated no green
cell. The downside? Their small size means they have appears. Researchers are hoping for “no glow” or in lab-
the potential to also activate many off-target molecules. speak, a “blank.” Blanks are the winners that go on to the
This collateral damage increases the chance of next stage of drug development.
adverse reactions.

ASSAY DEVELOPMENT:
GLOW AND NO GLOW
To find the best drug candidate, scientists need to
design easy-to-perform, large-scale, fast, and accurate
assays. Think automation. Think high throughput
screening. Scientists often work with tests that produce
a fluorescent signal or color change because color is easy
to measure, relatively inexpensive to design, and safe.
Depending on the assay, a color change may or may not
indicate a drug candidate works.

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HIT TO LEAD LEAD TO CANDIDATE
High throughput screening ideally results in several Biopharma companies typically put lead compounds
promising drug candidates or “hits.” To become “leads” through several rounds of optimization before they can
that merit animal testing, hits must pass rigorous in become drug candidates for preclinical testing. Lead
vitro testing. On this leg of the drug discovery voyage, optimization is the process by which a drug candidate
researchers ask three questions: is designed via iterative rounds of synthesis. For small
1. Is the hit safe? molecule drugs, this tweaking may include creating
derivatives of the lead. Often made using computer-
2. Is the hit specific to the target?
aided design (CAD), derivatives can help ensure the lead
3. Does the hit show promise to treat the disease? fits well with its target. Researchers then test each for
If a potential lead is supposed to inhibit an enzyme—it improved potency, fewer side effects, solubility (making
should act on that particular enzyme. A lead with effects sure that the drug stays liquid and won’t form clumps)
that are too general may result in a drug with dangerous and shelf life, otherwise known as stability.
adverse reactions. Therefore, researchers aim to develop Of course, optimization for large molecule drugs differs
a drug that inhibits the target at a concentration low because biologics are genetically encoded. Researchers
enough to avoid interfering with related enzymes. This have developed techniques to create mutations in
approach provides a therapeutic window within which proteins at specific locations. Again, the goal is to see if
the drug is effective and safe. changes produce a better drug—maybe even one up to
Huey Lewis’s rigorous standards. If so, researchers have
at long last an official drug candidate, one ready for the
rigors of preclinical testing in animals.

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GENE THERAPY CURES Life Science Training from Industry Experts

THE PROMISE OF GENE The corrected version of the RPE65 gene helps repair
THERAPY UNFOLDS patients’ retinal health and vision.

In many ways, 2017 was the year of gene therapy in the ASTOUNDING IMPROVEMENT
United States. Patients and pharmaceutical companies
Clinical trial participants’ vision loss ranged from mild
celebrated the approval of not one, but three treatments
to severe. The trials included patients from ages four
for otherwise untreatable health conditions. Researchers
to forty-four. For 93% of them (27 out of 29), Luxturna
have been working on developing safe, effective gene
treatment improved patients’ visual function, as shown
therapies for three decades. Early trials were plagued
by a “multi-luminance mobility testing (MLMT) score.”
with safety issues. Consequently, the field remained on
The MLMT measures the ability to navigate an obstacle
hold until researchers could address those problems
course in low light.
with new and improved gene-delivery vehicles. This
Weekly explores these innovative approaches to
intractable illness and disease.
COST OF A ONE-TIME CURE
Luxturna is priced at $425,000 per eye. Most patients
THE FIRST APPROVALS need treatment in both eyes. Its expense comes in part
from the nature of the treatment as a one-time, cure.
The FDA approved the first gene therapies in the United
Spark Therapeutics is discussing payment options with
States last year --Chymera and Yescarta--both chimeric
insurers to help allay the sticker shock. One plan in
antigen receptor T-cell (CAR-T) therapies. They deliver
the works with Pilgrim Health, a large non-profit New
a gene to cancer patients’ white blood cells in order
England-based insurer, hinges on outcomes. Should
to program them to attack specific cancer cells. CAR-T
the treatment fail to meet the intended outcome at
treatments involve removing a patient’s white blood
certain intervals post-treatment, Luxturna will refund
cells, programming them to contain a cancer-destroying
the treatment’s cost. Another proposal in the works
gene, and then re-administering them to the patient.
would allow insurers to spread payments out over
TWO FIRSTS IN ONE multiple years.

Just before year’s end, the FDA approved a therapy TERM OF THE WEEK: VECTOR
for a rare form of inherited blindness. Developed by
Scientists have adapted some viruses to transport
Spark Therapeutics (Philadelphia, PA), Luxturna is the
therapies by tweaking them to target disease instead of
first gene therapy to target a genetic disorder. It’s also
causing illness. These souped-up microbes are known
first in another significant way. Unlike the new cancer
as viral vectors. They are the vehicles that make genetic
treatments, patients receive Luxturna directly, via sub-
therapy like Luxturna possible. The virus itself is simply a
retinal injection.
segment of genetic material— RNA or DNA—surrounded
The blindness treated by Luxturna is known as “biallelic by a protein coat. Proteins on the surface of the vector
RPE65 mutation-associated retinal dystrophy.” (the modified virus) target proteins on a patient’s specific
• Biallelic: Pertaining to both copies of a particular diseased or malfunctioning cell surface. These viral
gene (allele) vectors then incorporate DNA into the patient’s genome
• RPE65: A protein in the retina that helps convert light by “tricking” the patient’s cells into producing the DNA it
into the electric signals the brain interprets as sight delivered. Ultimately, this enables a patient to make the
functional protein that he or she lacked.
• Retina: Light-sensitive tissue in the eye
Over a decade ago, gene therapy stalled because of
• Dystrophy: Wasting of tissue
safety concerns. Researchers could not initially control
the insertion point of modified genes. In some cases,

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the introduced genes disrupted patients’ other genes, while there is some evidence that lentiviral vectors are
causing serious illness. Researchers have now developed better at targeting blood and central nervous system
vectors that allow them to more precisely target where tissues. Lentiviral vectors have a higher carrying capacity
a therapeutic gene goes into a person’s genome, making as well, so may be preferred for very large genes or in
the treatment much safer because it doesn’t interfere cases where the goal is to deliver more than one gene.
with the function of critical genes.
LOOKING AHEAD
Luxturna’s success raises hopes of cures for numerous
other diseases caused by a defect in a single gene,
including hemophilia, sickle cell anemia, Huntington’s
disease, and other types of hereditary blindness. As
the table below indicates, many companies have gene
therapy treatments in clinical trials.

Types of Gene Therapy Vectors

The most commonly used type of viral vector for gene

therapy applications are adeno-associated viruses (AAV).
However, AAVs aren’t the only vector in town. Lentiviral
vectors are also being tested in gene therapy clinical
trials. The choice of vector depends in part on the target
tissue and the size of the genetic payload. AAV vectors so
2018 may see more groundbreaking approvals as these
far seem to be best at targeting eye and muscle tissues,
treatments make their way through clinical testing.

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DRUG DISCOVERY 301 Life Science Training from Industry Experts

DRUG DISCOVERY 301 • The time it takes to absorb a drug into the body;

Biotech Primer Weekly wrapped up last year by exploring • Maximum concentration (Cmax) of a drug in plasma
the first two stages of drug discovery. We looked at and target tissues;
how pharmaceutical companies identify drug targets, • Bioavailability (where in the body and at what
or the molecules (usually proteins) involved in an illness concentration the drug ends up);
that an ‘as yet undeveloped drug’ will hopefully act on. • Half-life, or how long a drug takes to reach half its
Next, we examined how researchers develop those maximum concentration in the body;
pharmaceutical candidates. Now we turn to what comes
• Clearance, or the time for a drug to reach
next for fledgling drugs on the arduous journey to FDA
undetectable levels through excretion.
approval. First, though, a smidgeon of Latin.
Pharmacodynamic analyses involve observing the
PRIMUM NON NOCERE biological repercussions of increasing amounts of a drug.
This maxim from Hippocrates means “First, do no harm.” Negative side effects include nausea, loss of appetite,
Before testing a drug on people, researchers must make fatigue, skin sensitivity, and changes in blood pressure
sure it’s safe in two important ways. First, they assess a or heart rate. Animal PK and PD studies give scientists an
substance’s safety in vitro. This Latin phrase means “in idea of what a safe and effective dose of a drug might be
glass”—that is, lab-grown cells. The cell type varies, but in people.
if possible, researchers use one that is relevant to the
disease in question. For example, they may use lung cells
to test a drug for respiratory syncytial virus (RSV).
In vivo testing (“in a living thing”) comes next. This area
of preclinical testing assesses a drug candidate’s toxicity
in at least two different species of animals, such as mice
and guinea pigs. The animals receive more of the drug
for longer than would human volunteers. Meanwhile,
lab technicians watch the animals for adverse effects.
Preclinical testing must follow the FDA’s Good
Laboratory Practice (GLP) guidelines. These regulations
help ensure scientific integrity and humane treatment of
laboratory animals. HUNTING FOR MUTANTS
Most drug candidates also undergo mutagenicity
HOMING IN ON THE SWEET SPOT
testing, which determines their likelihood of triggering
Almost any substance–even water!–can be toxic in mutations. Causing mutations indicates that a fledgling
very high amounts. Consequently, drug developers drug may be carcinogenic, hence often consigning it to
aim for just the right dose–one that gives the desired the graveyard of pharmaceutical failure. One common
effect with minimal unwanted consequences. Finding screening for mutagenicity is the Ames test, which
the sweet spot is the domain of pharmacokinetic identifies chemicals that cause increased rates of
(PK) and pharmacodynamic (PD) analyses. Think of mutations in bacteria.
pharmacokinetics as “what a body does to a drug” and
Researchers can also assess a drug’s carcinogenic
pharmacodynamics as “what a drug does to a body.”
potential by examining test animals for tumors.
PK analyses typically measure:

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Q/T TESTING intended drug target. More off-target interactions
mean a greater chance of undesirable side-effects.
Heart problems aren’t just bad for people, they’re
bad for baby drugs too. So, companies try to rule out THE PAPER CHASE
anything with potential cardiac side effects as early as
Once researchers amass enough safety data to ensure
possible. To find drugs that may be heart-unhealthy,
that a drug candidate will be safe for people, they submit
researchers look at the “QT interval.” This is the time
an Investigational New Drug (IND) application to the FDA.
between the start of the Q wave and the end of the
The IND application contains short-term safety data,
T wave in the heart’s electrical cycle. A lengthened
information on manufacturing the drug, and details
QT interval suggests improper activity in a person’s
about the methodology and design of clinical testing.
ventricles and is a risk factor for sudden death. If an
If the FDA blesses the application, the drug candidate
experimental drug binds to proteins on heart cells that
may enter Phase 1 clinical testing. Animal testing often
enable the flow of calcium ions into the heart, they may
continues in parallel with clinical (human) studies to
extend a QT interval. Longer intervals generally seal a
collect long-term safety data, especially for medicines
drug’s doom.
that treat chronic diseases.
ALTERNATIVES TO ANIMAL TESTING
Currently, drug development necessitates in vivo testing
to best understand how drugs act in a human body.
In vitro testing simply can’t replicate Home sapiens’
complex physiology. However, two innovations may
help reduce the animal testing involved in the search for
new treatments:
• 3D tissue arrays: Companies such as Organovo (San
Diego, CA) create tissue arrays that better mimic
human physiology than flat layers of cells in tissue
culture flasks.
• Computer modeling: Researchers at the University
COCKTAIL FODDER
of California, San Francisco, and SeaChange
Pharmaceuticals (San Francisco, CA) have For every 5,000 drugs tested preclinically, only about
developed software that reliably predicts small five show enough promise to justify submitting an IND
molecule drugs’ interaction with “off-target” application to the FDA. Once a drug clears the preclinical
molecules. These are cellular proteins other than the testing hurdle, it can move on to human testing. Tune in
next week to learn more about this crucial process.

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FROM DRUG DEVELOPMENT TO
APPROVAL: PHASE I/II Life Science Training from Industry Experts

PHASE I AND II CLINICAL TRIALS typically one hundred or fewer. In some cases, Phase I
trials may use patients rather than healthy volunteers.
Every drug in clinical use today, from the latest CAR-T
For example, cancer drugs have a level of toxicity that we
treatment to older cholesterol-lowering statins,
would not want to expose healthy volunteers to, but that
share one thing in common: they have all successfully
toxicity is an acceptable risk for patients who may have
navigated the rigorous clinical trials process. This is no
no other options.
small feat, as only ~10% of the drugs that enter Phase
I testing successfully emerge as marketed products. This first stage involves a number of different tests.
Those few drugs that show remarkable success in early First, volunteers take escalating doses of the drug under
clinical trials make headlines, and deservedly so. Recent close observation. If and when they experience adverse
drugs that have made a big splash include Amgen’s effects, they stop taking the drug. This establishes the
oncolytic virus drug Imlygic, which is restoring faith in Maximum Tolerated Dose, or MTD, which becomes a
therapeutic cancer vaccines, and AbbVie’s Venetoclax, benchmark for the remaining trials. The MTD helps
which seems to melt away tumors in chronic lymphocytic assure investigators and subjects that the treatment is
leukemia patients. unlikely to be toxic.

This week, we’ll take a look at the first two phases of Other studies look at pharmocodynamics (PD) and
clinical trials. pharmacokinetics (PK). The first examines what the drug
does to the body; the second, what the body does to the
TERM OF THE WEEK: ENDPOINT drug. These investigations help determine drug dosage.
Variations in how people of different sizes, ages, and
Clinical trials measure endpoints, that is, major health
genetic backgrounds etc., will likely respond to a new
outcomes. There are generally two types:
drug make testing in different populations critical.
• Clinical endpoints refer to benefits such as
survival, decreased pain, the absence of disease, or PHASE II
greater mobility.
Phase II examines drug efficacy as well as continuing
• Surrogate endpoints substitute for clinical safety tests. Phase II trials involve larger groups of
endpoints when they are impossible or impractical participants, all of whom are patients. Group size varies,
to measure. For instance, a clinical benefit, such depending on the target market. A drug being developed
as survival, hopefully, takes decades to observe. for Type 2 diabetes needs far more participants than one
Researchers may instead look to shorter-term for a rare disease such as ALS.
phenomena. For example, in studying a drug
Phase II studies are usually randomized, double-blind
designed to prevent heart disease, they can monitor
studies. This means that patients are randomly assigned
cholesterol levels instead of decreased fatality from
the drug or the placebo and even the researchers
heart attacks. Similarly, in some cancer treatments,
involved don’t know who receives which. This helps
reduced tumor size stands in for longer life.
guard against bias in determining who goes into which
The FDA requires that clinical protocols clearly define group. Otherwise, investigators may naturally want
endpoints. They are front and center in the application to ensure the sickest patients receive the promising
companies submit to the FDA that request permission experimental treatments.
to study a new drug. This application is called an
A placebo is also known as the “standard of care group”.
Investigational New Drug application or IND.
According to the National Institutes of Health (NIH),
the standard of care is a treatment accepted by medical
PHASE I experts as proper for a certain disease and widely used
The FDA divides clinical studies into three main phases. by medical professionals. For example, in clinical trials
Phase I usually tests drug safety in healthy volunteers, for Amgen’s Repatha, researchers compared its safety

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and efficacy against statins, the current standard of care COCKTAIL FODDER
for cholesterol reduction. Repatha was demonstrated to
The first antibiotic, penicillin, was discovered in 1928
be as effective as statins, and had a better safety profile
when microbiologist Alexander Fleming noticed the mold
in some patients, which led the FDA to approve Repatha.
Penicillium secreted a substance that killed his bacterial
THE POWER OF POSITIVE THINKING cultures. Inspired by this story, biochemist Akira Endo,
who was working at Sankyo Pharmaceuticals (Tokyo)
Investigators measure drug efficacy by monitoring
in 1972, decided to screen Penicillium cultures in hopes
predefined endpoints. They always look at effectiveness
of finding a chemical that would inhibit cholesterol.
relative to a control group--people who receive either
Endo’s quest paid off – he discovered a compound that
standard of care treatment or a placebo.
eventually went on to be one for the first statin drugs
The “placebo effect” may stem from an expectation or approved in 1987.
belief that the “medicine” will work. The placebo control
Why would mold cells secrete an anti-cholesterol drug?
is very important since scientists have firmly established
For the same reason they secrete penicillin – to kill the
that placebos can significantly affect patient health.
bacteria that they are competing with for resources.
Recent evidence shows that people dealing with chronic
Cholesterol is required for bacteria to build functional
pain can produce natural opioid painkillers in response
cell walls.
to a placebo. More rarely, the reverse of the placebo
effect – the “nocebo effect” – occurs. In other words, Next week, we’ll continue along the pathway to approval
some patients expect a study drug will either not help or as we zoom in on drug development Phase III and IV.
in fact harm them.

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FROM DRUG DEVELOPMENT
TO APPROVAL: PHASE III Life Science Training from Industry Experts

PHASE III IS NO GUARANTEE a new cholesterol medication, researchers might take

baseline cholesterol levels.
Our last Biotech Primer WEEKLY explored the riskiest
part of the human clinical trials pathway: Phase II. A crossover study’s strength lies in its ability to capture
About 70% of drugs that enter Phase II never make it differences in response to a drug versus a placebo in the
out. Most often, it’s because they fail to demonstrate same patient. This eliminates the inevitable individual
effectiveness. Even making it to Phase III is no guarantee variations among subjects in the experimental and
of success – about 40% of drugs fizzle out during this control groups in the more traditional parallel study.
period. In today’s WEEKLY, we look at the final stage of
clinical testing and the innovative clinical designs that are
pushing for faster drug approvals.

CHOICES, CHOICES, CHOICES

Phase III clinical trials continue to test a treatment’s
efficacy and safety, but in still larger groups of patients.
Bigger groups mean more statistically significant
results. As in Phase II, the trials have traditionally been
randomized, double-blind studies. If the investigational
drug appears to work, patients are allowed to continue
taking the medication after the trial ends, before
regulatory approval. This is an example of what doctors THE NEW KID IN TOWN:
and other medical professionals call compassionate use. ADAPTIVE DESIGN
Choices, choices, choices... In addition to randomized, Adaptive studies offer more flexibility than traditional
double-blind trials, researchers have other study designs designs, and are gaining in popularity as more
at their disposal, which we will explore below. efficient in bringing new drugs to the market. They
allow investigators to modify the trial design as they
THE PATRIARCHS: PARALLEL go, rather than spending time and money pursuing
VS CROSSOVER DESIGNS drug formulations or dosages that ultimately prove
Parallel studies use the classic experimental design. ineffective. For example, researchers may separate
Participants are divided into two groups. The treatment participants into different dosage groups. At a pre-
group receives the experimental drug. The control group specified time, they may note how patients respond to
receives either the current standard of care or a placebo. different doses. If one dose seems more effective, the
researchers will conduct the rest of the trials using only
Investigators sometimes decide to conduct a crossover
that dosage. Researchers set the trial protocol, which
study. Here, both groups receive the medicine and then
lays out the adaptation schedule and processes before
the placebo or vice versa. The treatments are separated
the trial begins.
by a washout period in which patients are taken off the
study medicine (or placebo) to eliminate any effects. One prominent example of adaptive design is I-SPY 2
study (Investigation of Serial Studies to Predict Your
Both parallel and crossover studies include a baseline
Therapeutic Response with Imaging and Molecular
period before patients take the drug or placebo. It allows
Analysis). This trial, being conducted by 20 different
researchers to gather the initial health information
cancer research centers across the US, examines up to
against which they will compare changes observed
12 different breast cancer therapies at once. Drugs that
during or after the study. For example, when assessing
show promise proceed to Phase III, allowing space for
other drugs to enter the trial. “Graduates” of the I-SPY 2

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study include AbbVie’s (North Chicago, IL) Veliparib and NEW APPROACH TO FIGHTING
Merck’s (Kenilworth, NJ) MK-2206. CANCER: UMBRELLA DESIGN
The 21st Century Cures Act promotes the use of These trials reflect the fact that scientists classify most
adaptive trial design. This benefits volunteers, as cancers as specific subtypes based on the mutation
researchers assign more patients to study groups taking involved. An umbrella study may look at patients with
medicine that demonstrates promise. It’s good news for one type of cancer, for example, lung cancer. However,
non-study patients too, who stand to benefit from new researchers divide participants into subgroups based
treatments getting to market faster. on the particular mutation behind their lung cancer, and
treat them with drugs designed to target the subtype
NEW APPROACH TO FIGHTING specifically. This allows researchers to identify patients
CANCER: BASKET DESIGN most likely to benefit from the new drug.
The design of clinical trials for new oncology drugs is
One ongoing umbrella study is the National Cancer
changing to reflect our increasing understanding of
Institute’s Alchemist Lung Cancer Enrichment Marker
cancer as a disease driven by mutations. Traditionally,
Identification and Sequencing Trials (ALCHEMIST) study.
trials test drugs on a specific illness, say, lung or
In it, researchers sequence patients’ tumors and assign
breast cancer. In contrast, basket trials classify cancers
them to different treatment groups according to the
according to the mutation they exhibit, not the tissue
subtype mutation.
they affect. For example, the FDA initially approved the
drug Zelboraf for melanoma patients with a mutation
in their “BRAF” gene. Researchers at Memorial Sloan
Kettering Cancer Center (New York, NY) explored
whether Zelboraf could work against other cancers with
the same genetic signature – the BRAF mutation. The
results indicated that Zelboraf also effectively treated
BRAF-mutated non-small cell lung cancer, Erdheim-
Chester disease, and Langerhans cell histiocytosis. These
last two, rare blood cancers, illustrate a key benefit of
basket trials: they may offer patients with rare cancers
better access to clinical trials, depending on their tumor’s
genetic signature.
Phase III is the final hurdle for drug candidates. If a
drug “survives,” things are looking good for developers
and patients alike. It’s not a slam dunk yet though, so
next week we look at the approval process and what
comes next.

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FROM DRUG DEVELOPMENT TO
APPROVAL: PHASE IV Life Science Training from Industry Experts

PHARMA FINISH LINE: FDA APPROVAL The FDA’s special designations include:

Last week, we focused on the final stage of clinical Accelerated Approval allows drugs to go forward
testing, Phase III trials, where drug developers assess the using surrogate endpoints instead of clinical endpoints.
safety and efficacy of their drug in large patient groups. Surrogate endpoints, such as lowered blood pressure
At the end of Phase III, drug developers face the moment or reduced tumor size predict rather than demonstrate
of truth: does the study data support claims that the clinical benefits. In these cases, pharmaceutical
new drug is both safe and effective? If the answer is yes, companies must run post-market studies to verify the
then it’s time to submit either a New Drug Application anticipated effect.
(NDA) for small molecule drugs, or a Biologics Licensing Priority Review means the FDA will aim for a decision
Application (BLA) for large molecule drugs. This week, within six months.
we’ll examine the approval process, including various Fast Track is based on preclinical or clinical data that
pathways for expedited approval, and touch on post- suggests the product addresses a specified unmet
approval safety studies also known as Phase IV. medical need. The designation enables developers to
communicate more often with the FDA. The agency
UNDER THE FDA MICROSCOPE provides guidance on clinical trial design and process,
Once a company submits either an NDA or BLA, the which helps resolve questions or issues quickly. These
FDA takes about a year for review–which seems designees also qualify for Accelerated Approval, Priority
governmentally slow unless you consider that most Review, and Rolling Review—which allows developers
applications run 100,000 pages or longer. There are to submit each section of an NDA or BLA as they finish,
three possible responses: rather than all at once.
• Approval letter: Ta dah! Breakthrough Therapy designates drugs that may
• Approvable letter: Close, and this is how to get the greatly improve patient health. The bar is set high
cigar! FDA requests correction of minor deficiencies, to join this privileged group, though. It requires
labeling changes, or post-approval studies. preliminary clinical evidence of effectiveness. Once
granted, Breakthrough designees receive Fast Track
• Not approvable letter: Thanks for playing! FDA
advantages, as well as intensive guidance on their
elaborates on deficiencies in the application and why
development program as early as Phase I. They also get
the drug is not approved.
an “organizational commitment involving senior FDA
For many drugs, the development, testing, and
managers,” according to the FDA website.
approval journey is straightforward. Difficult,
but straightforward. Orphan Drugs are often found in the above categories.
Companies develop them for rare diseases, those which
THAT SPECIAL SOMETHING affect fewer than 200,000 Americans. These include
hemophilia and Gaucher’s Disease, a genetic disorder
Certain drugs are given special consideration throughout
that causes skeletal and neurological issues. Prior to
the FDA approval process. These particular therapies are
the Orphan Drug Act of 1983, the industry had no
eligible for regulatory designations that speed up the
financial incentives to work on therapies for such small
review and get a product to market more quickly. They
populations. Orphans lacked commercial sponsors,
must first meet two criteria. A drug must target a serious
“parents,” to shepherd them through lengthy and
condition that likely results in death or significantly
expensive trials. The orphan drug legislation provides
impairs daily living. Examples include cancer and
the following incentives:
Alzheimer’s disease. The drug also needs to address a
major, unmet medical need. That is, either no medicine • Federal tax credits of up to 50% off research costs
exists, or current therapy has safety issues. • Increased protection from generic competition

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 • Waivers of FDA application and product fees quickly reveal whether a patient can benefit from Plavix.
(amounting to hundreds of thousands of dollars) This unlooked-for result spurred the FDA to require that
the Plavix label carry a black box warning – a warning
COCKTAIL FODDER: PDUFA outlined in a black box, which is the strictest warning put
The Prescription Drug User Fee Act (PDUFA) requires on the labeling of prescription drugs or drug products by
drug developers to pay a fee to the FDA to help fund the the FDA.
work necessary for timely approvals. The “PDUFA date” The rigorous path from preclinical testing to approval
of a drug is the date by which the FDA has committed to usually takes a decade or longer, with no promise of
review its application. For 2018, the PDUFA fee required success. According to the Biotechnology Innovation
when submitting an NDA or BLA that requires clinical Organization, approximately one in ten drugs entering
data is $2,421,495. clinical testing ultimately make the grade. The highest
rate of failure occurs in Phase II; only 31% of drugs
FINISHED, NOT DONE proceed to Phase III. Of those that make it to the final
Phase IV generally refers to post-market studies, which Phase III, a little more than half - 58% - are considered
companies undertake after a drug is approved and at the safe and effective enough for an NDA or BLA submission.
pharmacy. Drugs that were approved using a surrogate From start to finish, only 10% of the drugs that begin
endpoint are monitored to confirm clinical efficacy. For Phase I reach the market.
all drugs, safety is monitored and confirmed. Phase IV
includes informal studies of doctor and patient reports,
which sometimes reveal unanticipated side effects.
Such “surprises” aren’t uncommon in brand new drugs,
because so many more people take them once out
on the market versus the small number of patients in
clinical trials.
For instance, after the anti-clotting medicine Plavix
(Bristol-Myers Squibb; New York, NY) was approved,
doctors found it less effective in some people than
expected. Further probing found that the drug wasn’t
fully activated in about 14% of patients because their
Despite the low odds, innovative companies continue to
bodies produced a less active form of the liver enzyme
bring new drugs to market every year. Stay with us as we
that activates Plavix. Fortunately, a genetic test can
continue following their stories in 2018.

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MARKET ACCESS Life Science Training from Industry Experts

MARKET ACCESS PRIMER Outcomes-based contracts include some sort of refund

if the new drug does not perform as advertised. New
For the last few weeks we here at Biotech Primer have
England-based insurer Harvard Pilgrim’s contract
tracked the progression of a drug candidate from the
with Amgen (Thousand Oaks, CA) provides a “pay-for-
lab to the marketplace, where only the fittest survive.
performance” rebate to Harvard Pilgrim if Amgen’s
Winning at clinical trials means earning an official
new cholesterol drug Repatha, a PCSK9-inhibitor drug,
regulatory approval. Congratulations! But as any
doesn’t perform as well as in clinical trials. The benefit to
seasoned drug developer will tell you, the game has only
Amgen is preferred positioning on the Harvard Pilgrim
just begun. Ensuring newly-approved drugs are available
formulary as well as access to real world and post-
to all patients is the next hurdle.
marketing data.
In this WEEKLY we’ll explore market access for
With other novel high-impact, high-cost drugs such
innovative products such as cancer-fighting drugs and
as CAR-T treatments and gene therapies, payors are
gene therapies.
questioning how to best afford these medicines. For
example, Spark Therapeutics (Philadelphia, PA) recently
TERM OF THE WEEK: MARKET ACCESS received FDA approval for Luxturna, a gene therapy
Market access is the process of proving a product’s treatment for hereditary blindness. The treatment is
value to ensure reimbursement with commercial payors, remarkable, in that it promises a lifetime cure with one
government payors and integrated delivery systems. administration - at a cost of $425,000 per eye. Spark
These payor stakeholders are often referred to as Therapeutics is working on an outcome-based payment
“organized customer groups.” scheme in collaboration with Harvard Pilgrim Health
According to Linda Lander, President of Inside Out and with Centers for Medicare and Medicaid Services
Market Access, “market access aligns incentives between (CMS). If Luxturna fails to meet the intended outcomes
payors, pharmaceutical companies, providers, and at certain intervals post treatment, Spark would refund
patients to lead to cost-effective models.” some or all the treatment’s cost.
These types of agreements allow patients the chance to
DRUG APPROVAL DOESN’T benefit from a new therapy that a payor might otherwise
EQUAL REIMBURSEMENT be reluctant to take a chance on.
Launching a new drug, especially a high-cost one,
requires proving its value early in the process. Just AN INDEPENDENT VOICE
because a drug is approved doesn’t mean it will be Organized customer groups are increasingly using
reimbursed for coverage. Without reimbursement independent analysis to guide reimbursement decisions.
patients will not benefit, nor will the drug’s sponsor These independent agencies, which include the
recover the significant resources it invested California Technology Association (CTA), the National
in development. Comprehensive Care Network (NCCN), and Institute
Organized customer groups are now demanding of Clinical and Economic Review (ICER), evaluate and
evidence of value. Market access involves a discussion determine the “reasonable price/value” of drugs in the
and negotiation between the payor and the drug context of an overall healthcare budget.
sponsor to define and demonstrate value. This is According to Steve Pearson, Executive Director of ICER,
increasingly achieved via outcome-based reimbursement the organization asks four key questions:
and real world evidence, in which patient health and
• How well does the drug work?
outcomes data gathered outside of clinical trials drives
downstream reimbursement. • How much better is it than what we already have?
• How much could it save us?

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 • How much would it cost to treat everyone who As a result, biopharma companies increasingly work with
needs it? ICER and other independent organizations to gain payor
In conducting its analysis, ICER reaches out to patients, acceptance for their initial drug prices; for example,
doctors, and drug makers. The end goal is to calculate Spark Therapeutics met with ICER prior to the Luxturna
a value-based price benchmark that reflects the launch to help guide pricing strategy.
drug’s value to patients and to the healthcare system. As new technologies create highly innovative treatments,
According to the American Managed Care Pharmacy’s the issue of pricing must be addressed. By deftly
annual survey of payors, more than 50 percent now navigating market access, companies can ensure more
employ ICER evaluations in their reimbursement reviews. patients have access to these advances.

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FROM DRUG DEVELOPMENT
TO APPROVAL: A RECAP Life Science Training from Industry Experts

FROM THE LAB TO THE PATIENT 3. Does the hit show promise to treat the disease?

In this issue of the Biotech Primer WEEKLY we will Biopharma companies typically put lead compounds
recap the past seven issues that highlight the journey a through several rounds of optimization before they can
molecule takes from the lab to the patient. become drug candidates for preclinical testing. Lead
optimization is the process by which a drug candidate is
Beginning in the 1980’s, scientists took a new tack in
designed via iterative rounds of synthesis.
developing drugs. They adopted an approach known
as rational drug discovery. Using this methodology, Drug Discovery 201, published December 2017
researchers first seek to understand a disease at the
cellular level to identify its mechanism. Once the cellular
DRUG DISCOVERY: SAFETY
mechanism becomes clear, scientists can identify a drug
AND SUBMISSION
target: the molecule involved in the illness that the ‘as In vivo testing (“in a living thing”) comes next. This area
yet undeveloped drug’ will hopefully act on. of preclinical testing assesses a drug candidate’s toxicity
in at least two different species of animals, such as mice
Fast forward many years and hundreds of millions of
and guinea pigs. The animals receive more of the drug
dollars and drum roll please... the ‘as yet undeveloped
for longer than would human volunteers. Meanwhile,
drug’ may just become the drug that cures your once
lab technicians watch the animals for adverse effects.
incurable illness.
Preclinical testing must follow the FDA’s Good
DRUG DISCOVERY: IDENTIFY Laboratory Practice (GLP) guidelines. These regulations
AND VERIFY help ensure scientific integrity and humane treatment of
laboratory animals.
Researchers identify their molecular targets by
answering one incredibly complex question: How does Once researchers amass enough safety data to ensure
diseased tissue differ from healthy tissue? that a drug candidate will be safe for people, they submit
an Investigational New Drug (IND) application to the FDA.
Researchers verify the suitability of target molecules by
If the FDA blesses the application, the drug candidate
answering two important questions: does the target play
may enter Phase 1 clinical testing.
a key role in the disease? and will targeting the molecule
likely be safe and effective? Drug Discovery 301, published January 2018

Identifying and validating drug targets are only the first PHASE I/II: MAY THE ODDS
steps on the long road to safe, effective new medicines. BE IN YOUR FAVOR
Drug Discovery 101, published December 2017 Every drug in clinical use today, from the latest CAR-T
treatment to older cholesterol-lowering statins,
DRUG DISCOVERY: HIT TO LEAD share one thing in common: they have all successfully
To find the best drug candidate, scientists need to design navigated the rigorous clinical trials process. This is no
easy-to-perform, large-scale, fast, and accurate assays. small feat, as only ~10% of the drugs that enter Phase
Think automation. Think high throughput screening. I testing successfully emerge as marketed products.
High throughput screening ideally results in several Those few drugs that show remarkable success in early
promising drug candidates or “hits.” To become “leads” clinical trials make headlines, and deservedly so.
that merit animal testing, hits must pass rigorous in Phase I/II, published January 2018
vitro testing. On this leg of the drug discovery voyage,
researchers ask three questions: PHASE III: CHOICES, CHOICES, CHOICES
1. Is the hit safe? Phase III clinical trials continue to test a treatment’s
2. Is the hit specific to the target? efficacy and safety, but in still larger groups of patients.

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Bigger groups mean more statistically significant Review, and Rolling Review—which allows developers to
results. As in Phase II, the trials have traditionally been submit each section of an New Drug Application (NDA) or
randomized, double-blind studies. If the investigational Biologics License Agreement (BLA) as they finish, rather
drug appears to work, patients are allowed to continue than all at once.
taking the medication after the trial ends, before Breakthrough Therapy designates drugs that may
regulatory approval. This is an example of what doctors greatly improve patient health. The bar is set high
and other medical professionals call compassionate use. to join this privileged group, though. It requires
Choices, choices, choices… In addition to randomized, preliminary clinical evidence of effectiveness. Once
double-blind trials, researchers have other study designs granted, Breakthrough designees receive Fast Track
at their disposal, which we will explore below. advantages, as well as intensive guidance on their
Phase III, published February 2018 development program as early as Phase I. They also get
an “organizational commitment involving senior FDA
PHASE IV: THAT SOMETHING SPECIAL managers,” according to the FDA website.

The FDA’s special designations include: Phase IV, published February 2018

Accelerated Approval allows drugs to go forward

GETTING MEDICATIONS TO
using surrogate endpoints instead of clinical endpoints.
PATIENTS: MARKET ACCESS
Surrogate endpoints, such as lowered blood pressure
or reduced tumor size predict rather than demonstrate Market access is the process of proving a product’s
clinical benefits. In these cases, pharmaceutical value to ensure reimbursement with commercial payors,
companies must run post-market studies to verify the government payors and integrated delivery systems.
anticipated effect. These payor stakeholders are often referred to as
“organized customer groups.”
Priority Review means the FDA will aim for a decision
within six months. According to Linda Lander, President of Inside Out
Market Access, “market access aligns incentives between
Fast Track is based on preclinical or clinical data that
payors, pharmaceutical companies, providers, and
suggests the product addresses a specified unmet
patients to lead to cost-effective models.”
medical need. The designation enables developers to
communicate more often with the FDA. The agency Market Access, published February 2018
provides guidance on clinical trial design and process, Despite the low odds, innovative biopharma companies
which helps resolve questions or issues quickly. These continue to bring new drugs to market every year. Stay
designees also qualify for Accelerated Approval, Priority with us as we continue following their stories in 2018.

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WHO’S YOUR DADDY? THE
SCIENCE OF 23ANDME Life Science Training from Industry Experts

HEY—CHECK OUT THOSE GENES! “STR” stands for “short tandem repeat.” These are short
sequences of DNA that repeat from five to fifty times.
There’s an old saying, “If you don’t know where you’ve
The number of times a particular STR repeats varies
come from, you can’t know where you’re going.” We used
from person to person.
to rely on paper birth certificates, marriage licenses and
memory to help discover where we’ve come from; but THIS DNA IS NOT THAT DNA
paper gets damaged, people are fallible, and memories
SNPs and STRs provide different types of information
fade. Leave it to biotech to come up with a better way.
depending on their DNA origin. We humans have
New heritage-hunting techniques come courtesy of one
different types of DNA:
our greatest scientific achievements: the sequencing of
the human genome. • The sex chromosomes are the X and the Y
chromosomes. Men have one Y chromosome and
We now have tools to genetically examine ourselves
one X chromosome; women have two Xs only. Thus,
and our past. The ability to decode genetics at modest
the Y chromosome contains DNA information about
expense allowed the DNA testing industry to develop.
paternal ancestry.
Companies such as 23andMe, Ancestry.com, and
National Geographic sell genetic tests online. These • Mitochondrial DNA (mtDNA) is found in tiny
kits, consisting of a plastic tube, a baggy, and a postage- compartments called mitochondria that convert
paid mailer, promise biological insight into the past. But sugar to energy in cells. Only woman pass on mtDNA
what about the science? This Biotech Primer Weekly because during fertilization the DNA in sperm
overviews commercial DNA testing and how it works. mitochondria is quickly destroyed. Mitochondrial
DNA can help trace maternal ancestry.
A DASH OF DIFFERENCE • Autosomal DNA is all the genetic material that is not
Every human being alive shares an astounding 99.9% found in the sex chromosomes and in mitochondrial
of their DNA! Only one tenth of one percent of genetic DNA. Autosomal DNA is found in the pairs of
“stuff” accounts for a planetful of differences. That’s autosomal chromosomes numbered one through
because our genome consists of an astonishing three 22. These 22 pairs come from both mom and
billion-plus building blocks—the famous As, Cs, Ts dad – one copy from each. Autosomal DNA offers
and Gs. generic information about both the maternal and
Areas of variation in genes are called genetic markers. paternal lineages.
Geneticists and others use genetic markers to assess the
probability that two people share a common ancestor.
DEEP ANCESTRY
The more genetic markers two people share, the more Not only are types of DNA inherited differently – some
likely they are related. The only people with the same from mom, some from dad, some from both – but
exact genetic markers are identical twins. the rate at which their DNA sequence changes from
generation to generation also varies. This is because
ALPHABET SOUP: SNPS AND STRS mtDNA and Y chromosomal DNA change only through
The two major types of genetic markers are SNPs and random mutation of their As, Cs, Ts and Gs. Change can
STRs. In the alphabet soup of genetics, “SNP” is short take centuries before a noticeable difference appears.
hand for “single nucleotide polymorphism.” It simply By comparing genetic markers from mtDNA and Y
means that one of the three billion building blocks that chromosomal DNA with those of indigenous peoples
make up a person’s DNA has been switched. In other from various parts of the world, it’s possible to estimate
words, an A has been swapped with a G. where your ancestors hailed from way back when.

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MAKING FAMILY CONNECTIONS each egg a woman produces only carries half her DNA
only some will carry Mongolian markers. Therefore, one
Each person shares half their genetic markers with
sibling could show Mongolian ancestry, while another
siblings, parents, and any children they have. We share
doesn’t.
about a quarter with grandparents or grandchildren,
aunts and uncles, nieces and nephews, and half-siblings. OUR POSSIBLY HAIRY PAST
The amount of DNA we have in common with relatives
What’s one of the most intriguing secrets genetic
diminishes by half in each successive generation. By
testing can reveal? Just how much of a Neanderthal you
seven generations back, the amount of DNA shared
really are! Our smaller, sturdier cousins’ genome was
among relatives is less than one percent.
published in 2010, based on some very well-preserved
So, autosomal DNA is useful in trying to make DNA taken from bones found in a Croatian cave. The
connections between living relatives or determining how evidence suggests that early humans mated with
far back you share an ancestor with someone. However, Homo neanderthalensis. Thanks to DNA testing, we
it doesn’t shed light on “deep ancestry,” or the region of can find out just how cozy our distant relatives were
the world our ancestors came from centuries or even with Neanderthals. The average person is about five
millennia ago. percent Neanderthal.

COCKTAIL FODDER: SAME FAMILY, Today’s technology makes it possible for the curious
DIFFERENT ANCESTOR? to shed light on their DNA. The databases of genetic
markers are growing apace, making it easier to uncover
Can siblings’ ancestral origins be different? The
long-lost relatives, for better or for worse. So how
surprising answer is yes! Imagine that a small
about it? Are you ready to find out who you really are,
percentage, say 10 percent, of a woman’s DNA contained
biologically speaking?
genetic markers indicating Mongolian ancestry. Because

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MEATLESS MEAT: BIOTECH BURGER
AIN’T NO BEAN PATTY Life Science Training from Industry Experts

MISSION IMPOSSIBLE: MEATLESS MEAT? Molecules with different masses travel at different
rates, creating a graph or mass spectrum. Researchers
Imagine biting into a juicy cheeseburger: the flavor,
compare their speeds to a baseline spectrum,
the texture, the smell. Now, imagine the cheeseburger
which allows them to detect the molecules in their
meatless. Impossible? No. Impossible Foods, a Redwood
sample burger.
City, CA-based company has used biotechnology to
create a plant-based burger amazingly similar to the THE HAMBURGER MOLECULE
bovine original. In this edition of the Biotech Primer
One key molecule Impossible Foods scientists identified
Weekly, we examine how they did it.
as responsible for a hamburger’s meaty flavor is heme.
UNCOVERING THE MYSTERY OF MEAT This iron-carrying molecule is associated with one of the
major muscle proteins, myoglobin. Since meat is pretty
Impossible Foods founder Patrick Brown knew he
much muscle, it’s no surprise that it’s pretty much heme
needed to understand America’s favorite sandwich at
that accounts for meat’s taste.
its most basic level. Discovering the specific molecules
that imbue hamburgers with their unique deliciousness, Heme isn’t merely some muscle-bound molecule. It’s
he reasoned, would enable him to build a kinder, more also found in the roots of soy plants, attached to the
planet-friendly burger. protein leghemoglobin. Soy contains far less heme
than cow muscle. That’s why soy-based veggie burgers
So how to get at what makes a burger a burger?
lack that signature meaty taste. Soy contains so little
Step one: acquire hamburger meat. vegetarian heme that using the plant to meatify veggie
Step two: cook. Researchers at Impossible Foods heated burgers just won’t work.
ground beef, essentially vaporizing it into a usable There’s more than one way to skin a soybean though—
sample. These gaseous burger components include those savvy Impossible Foods people genetically
some of the same molecules responsible for those engineered yeast cells to contain the soy leghemoglobin
mouth-watering smells that waft through neighborhoods gene. Voila! A sustainable, high-volume production
and parks in warm weather. platform for leghemoglobin is suddenly possible.
Step three of the molecular dissection is a little more Heme is harvested from the yeast to become a major
complicated than meat and heat. It requires a gas ingredient the secret Impossible Burger recipe. The
chromatogram mass spectrometer, which is really two recipe also includes wheat and potato proteins for
gadgets in one. Not standard kitchen equipment. texture, and coconut oil as a stand in for fat.

Chromatography is the process of separating

ENGINEERING THAT BLOODY TASTE
components of a mixture. Burger “gas,” like the earth’s
atmosphere, is composed of molecules with different Impossible Foods set out to recreate not only the taste,
physical and chemical properties. Think different sizes, but other parts of the burger-eating experience. That
shapes, and charges. Scientists pass the burger fumes leads to the question: does a medium rare Impossible
through a long, narrow tube that contains a material that Burger ooze a little “blood?”
various molecules “stick to” to a greater or lesser degree, Of course. The faux blood looks strikingly similar to the
based on their makeup. The assorted molecules leave real thing. That’s because heme not only makes meat
the tube at different rates, separating. taste meaty, it makes blood red. Heme binds iron, which
The exiting molecules then move on to the spectrometer in turn binds oxygen. The chemical bond between iron
which “ionizes” or gives them an electric charge. and oxygen reflects light so that it appears crimson.
Negatively charged molecules then pass through a While the editorial staff of the Weekly has yet to
magnetic field toward a negatively charged panel. sample the Impossible Burger (hint hint, Dr. Brown),

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eater-on-the-street reports are favorable. How does beef runs thousands of dollars. Miraculously, Kobe beef
it stack up nutritionally? Protein, fat, and iron content seems almost affordable.
are comparable to a lean ground beef patty – with the Several companies are working to make lab-grown
added benefit of zero cholesterol. The Impossible Burger beef cheaper and easier to produce. These include
is currently available in restaurants at select locations SuperMeat (Tel Aviv, Israel); Mosameat (Maastricht, The
around the country. Netherlands); and Memphis Meats (San Francisco, CA).
Memphis Meats expects to have a supermarket-ready
BEYOND IMPOSSIBLE: product by 2021; Mosameat estimates a similar timeline.
LAB-GROWN MEAT
Going vegetarian can be tough, but many find it a
But wait, there’s more! Another type of biotech burger is
pathway to better health. Moreover, beef production
on the horizon: lab-grown meat. Basically, researchers
uses significantly more natural resources than the
extract stem cells from cow muscle and grow them in
equivalent amount of veggie or cultured meat. And that’s
the lab.
not to mention all that beefy methane. Global livestock
This approach to making “cultured” or “clean” meat still production accounts for about 15% of this greenhouse
has a way to go before it lands on your dinner plate. gas, which contributes to global warming. For those
Growing cells in the lab isn’t cheap. Similar to animals, who want to cut back or eliminate meat consumption,
cultured meat cells need care and feeding. The health, but still crave the taste of a good old-fashioned burger,
safety and technological requirements of culturing biotechnology just might have the answer.
meat result in great expense. One pound of lab-grown

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ALZHEIMER’S DISEASE: A TOUGH NUT TO CRACK Life Science Training from Industry Experts

AFFECTING 5.1 MILLION TANGLES

Alzheimer’s disease (AD) ranks as one of the toughest Another brain protein associated with Alzheimer’s is tau,
nuts to crack within drug discovery and development. which is concentrated in the neurons and is primarily
Current treatments merely manage symptoms, so understood as a component in stabilizing nerve cell
finding a better solution becomes more and more urgent structure. Abnormal aggregates of tau form “tangles”
as the aging population grows. within nerve cells. These tangles are correlated with
Approximately 70 percent of dementia cases are caused the onset of Alzheimer’s along with the characteristic
by AD. It is a neurodegenerative disorder— neurons plaque formations.
progressively lose structure and function. As the disease
continues and more neurons are damaged and die,
GENETICS PLAYS A ROLE
symptoms get worse. Neurons in the hippocampal About 70% of Alzheimer’s cases are thought to have
region of the brain associated with memory formation at least some genetic association, with different genes
are among the first affected. By 2025, the number of being implicated depending on the type of Alzheimer’s.
people age 65 and older with AD is projected to reach A gene found on chromosome 19 called the
7.1 million—a 40 percent increase from the 5.1 million apolipoprotein E gene (APOE) influences the
affected in 2015 (Alzheimer’s Association). development of late-onset Alzheimer’s. Individuals
A number of different companies are working to develop with different variants of the APOE gene have different
treatments, with a few already in clinical trials. In this risk profiles:
two-part series, we’ll first explain what is known about • Variant ε2 (APOE2) is rare and possibly lessens or
the molecular causes of AD, then explore Alzheimer’s delays Alzheimer’s onset
drugs that are in development.
• Variant ε3 (APOE3) is neutral
PLAQUES AND TANGLES • Variant ε4 (APOE4) is associated with a significantly
increased risk of Alzheimer’s
PLAQUES The APOE proteins plays a role in clearing Aβ from
Alzheimer’s disease is associated with the build-up the brain, with APOE4 carrying out this function less
of amyloid-beta (Aβ) plaques in patients’ brains. But efficiently than the other variants. There is also some
what, exactly, are Aβ plaques? Aβ plaques derive from evidence that APOE4 contributes to the breakdown
the cleavage of a protein called the amyloid precursor of the blood-brain barrier seen in patients, resulting
protein, which is thought to play a role in the formation in increased brain inflammation—another marker of
of synapses. Individual Aβ molecules clump together to Alzheimer’s. A better understanding of APOE4’s role
form the plaques associated with Alzheimer’s. in Alzheimer’s onset may lead to the development of a
Until recently, the mechanism by which Aβ plaques whole new class of drug.
might cause Alzheimer’s was not known. Recent research
from Stanford University suggests the plaques bind
IF AT FIRST YOU DON’T SUCCEED…
to a receptor on nerve cells, disrupting their function. A number of drug developers have attempted to use
However, there is no absolute consensus on whether monoclonal antibodies (mAbs) to disrupt the formation
these clumps of protein are the origins of AD or a of the AD-associated Aβ plaques. Unfortunately,
symptom of the underlying cause. this approach has yet to experience clinical success.
However, this doesn’t mean that the approach is not
viable – different mAbs attach to different parts of Aβ,
meaning that the outcome of one mAb trial does not
necessarily predict the outcome of another. Biogen

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(Cambridge, MA) and Roche (Basel, Switzerland) both UNTANGLING TAU
have mAb therapies targeting Aβ plaques in Phase III
AbbVie (North Chicago, IL) is targeting tau, the other
clinical development. In earlier phase studies, these
major protein associated with AD. The company
mAbs showed the most promise in patients with less
currently has an anti-tau mAb therapy in Phase II clinical
advanced forms of the disease.
testing, and last month announced a partnership with
The prevention of Aβ plaques is also the focus of Voyager Therapeutics (Cambridge, MA) to develop a gene
growing interest in creating an AD vaccine. Leading this therapy treatment that targets tau. The treatment will
effort is Novartis (Basel, Switzerland), whose CAD106 deliver a gene encoding an anti-tau antibody directly to
vaccine contains fragments of the Aβ protein and has cells in patients’ brains – enabling those cells to make the
been shown to be safe in Phase I trials. The goal of the antibody. If successful, this would bypass altogether the
vaccine is to activate an immune response against Aβ, blood-brain barrier that makes it difficult for some drugs
thereby reducing its accumulation and potential to form to even enter the brain.
plaques in the brain. CAD106 is currently in Phase II/III
studies of efficacy, in which the vaccine is being tested THE FUTURE?
in cognitively normal individuals between the ages of
Amyloid-beta and tau remain tantalizing targets for AD
60 and 70 who are at high risk of developing the disease
drug development because of their close association
based on their APOE4 status.
with the disease. However, new approaches are
springing up as frustration over the lack of progress in
treating this devastating disease grows. Stay tuned for
the next Weekly, where we will describe some of these
new approaches.

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EXPLORING DIFFERENT STRATEGIES
TO FIGHT ALZHEIMER’S Life Science Training from Industry Experts

TAKE THAT, ALZHEIMER’S The new cells migrated to the correct location and
differentiated. Moreover, they survived long enough
Alzheimer’s pernicious amyloid-beta plaques and
to reverse previously observed cognitive declines. The
tau tangles, discussed last week, remain important
hippocampus is one of the first regions of the brain
targets for the biotech industry. In the past few years,
to show damage in AD and is thought to play a role in
however, companies have begun to search more broadly
memory formation and spatial navigation. Neuronascent
for new treatments. This Weekly looks at products in
is preparing for Phase I trials of NNI-362.
development that use different strategies to fight this
heartbreaking illness. NEUROINFLAMMATION
REVIVING THE BRAIN? Neuroinflammation is one of the drivers of
neurodegeneration in Alzheimer’s disease, multiple
Loss of neurons is Alzheimer’s signature, devastating
sclerosis and other brain disorders. Research conducted
effect. What if we could jump start the development of
at Stanford University (Palo Alto, CA) suggests that
new brain cells? Two companies are trying to do just that.
the protein c1q is present at higher levels in people
Neurotrope Biosciences (New York, NY) is developing with Alzheimer’s disease. C1q accumulates at neuronal
bryostatin, a drug that activates protein kinase C synapses, the key points of communication between
epsilon (PKCꞓ). This protein plays a key role in forming brain cells. This protein also signals other immune cells,
memories. In animal models of stroke, traumatic brain such as macrophages—which then chomp up cellular
injury, and Alzheimer’s disease, bryostatin appears to debris present in affected brains. The accumulation
restore deficits in synapses (connections between brain of c1q could account for the loss of synapses and
cells) and decrease cell death. These results suggest that accompanying mental decline.
bryostatin could help to prevent the loss of neurons and
South San Francisco-based Annexon is working
restore and generate new synapses.
on a promising therapy that centers on controlling
Phase II clinical studies of late-stage Alzheimer’s inflammation in the brain. ANX005, now in preclinical
patients demonstrated improved cognitive function as development, is a monoclonal antibody that mops up
measured by the Severe Impairment Battery Scale (SIB), excess c1q.
a standard tool for evaluating treatment response in
Another company homing in on neuroinflammation
advanced Alzheimer’s. Their improvement was greater
is vTv Therapeutics (High Point, NC). Their drug,
than that seen in patients given the placebo, but the
Azeliragon, now in Phase III clinical development, is a
difference was not statistically significant. A Neurotrope
small molecule inhibitor of the receptor for advanced
Biosciences spokesperson says that it considers the
glycation endproducts (RAGE). RAGE is present on many
Phase II study exploratory, designed to determine
neurological cell types. Its activation may promote
correct dosing. The company is planning a larger
amyloid-beta production and transport, tau aggregation,
confirmatory trial in hopes of demonstrating statistically
and chronic inflammation. Preventing any of these
significant efficacy.
developments could improve Alzheimer’s symptoms.
Neuronascent (Clarksville, MD) also aims to develop
small molecule activators of neurogenesis. By screening AND THAT AND THAT AND THAT!
large chemical libraries, the company has identified Rather than target Aβ plaques directly, Yumanity
compounds that show promise of sparking neurogenesis Therapeutics (Cambridge, MA) is trying to identify
from adult neural stem cells in both tissue culture and the problems they cause. Yumanity scientists have
mouse models. engineered yeast cells to overproduce the Aβ protein
The company’s lead compound, NNI-362, promoted and monitor its detrimental effects, such as disrupting
the growth of new hippocampal neurons in mice. the action of other important cellular proteins.

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Surprisingly, yeast share many molecular pathways with Oryzon researchers identified an enzyme, lysine-specific
humans. This similarity means researchers can use the histone demethylase 1 (LSD1), which makes epigenetic
little fungi to screen for potential drugs that address modifications to genes that results in “turning them
protein disruption. Promising candidates are then down” so they produce less of the corresponding
tested in Alzheimer’s patient-derived cells. By tackling protein. LSD1 makes these changes to genes that
a completely different disease mechanism, the new support neuronal survival. Oryzon scientists have
compounds may achieve greater success than seen so designed a drug, ORY-2001, that inhibits LSD1. Inhibiting
far with drugs that act directly on amyloid beta or tau. LSD1 could mean that more neurons survive in AD
Yumanity is currently in the lead-optimization phase of patients, leading to improved cognitive function. ORY-
pre-clinical development. 2001 recently entered Phase II clinical trials.
In partnership with Biogen (Cambridge, MA), Cambridge-
based Proteostasis Therapeutics is targeting
A TWIST ON TAU
AD-associated protein aggregates by activating Finally, the elusive AD treatment may lie in pursuing a
proteasomes. These cellular components get rid of well-established target after all—but at a new angle.
damaged proteins and dysfunctional protein aggregates That’s where Ionis Pharmaceuticals (Carlsbad, CA) is
by dismantling their chemical bonds. The protein USP14 headed in the Phase I/IIA clinical studies of their drug,
inhibits proteasomes. Proteostasis is working on the IONIS-MAPT. This antisense drug targets the source of
preclinical development of a USP14 inhibitor that allows the tangles associated with AD. Like other antisense
proteasomes to fully activate in AD patients. This makes drugs, IONIS-MAPT destroys tau mRNA, thereby
it more likely that the proteasomes will recognize and diminishing tau protein production.
destroy amyloid plaques and tau tangles. It’s encouraging to know how many therapies are in the
Oryzon Genomics (Barcelona, Spain) is taking an Alzheimer’s treatment pipeline. With more hard work
epigenetic approach to Alzheimer’s. Epigenetic and investment, perhaps one of the many introduced
modifications are chemical changes to gene sequences above will lead to a cure.
that don’t change the information content but instead
affect how much that content is used – in other words,
the amount of a particular protein that the body makes.

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CANCER DIAGNOSTIC IN A DROP Life Science Training from Industry Experts

THE LATEST IN CANCER DIAGNOSTICS Genomic Health (Redwood City, CA) currently markets
tissue-based genomic tests for the detection and
Hearing the words “it might be cancer” paired with
classification of breast and prostate cancer, and is
your doctor’s perplexed look is enough to send shock
developing cfDNA-based tests for breast (blood sample)
waves through your body. Getting to the heart of a
and bladder (urine sample) cancers.
diagnosis usually requires a surgical biopsy—removal
and examination of the suspected tissue for visible signs Qiagen (Hilden, Germany) is developing cfDNA liquid
of cancer. biopsy diagnostics in partnership with pharmaceutical
companies such as AstraZeneca (London, U.K.), Tokai
Less invasive diagnostic tests—called liquid biopsies—
Pharmaceuticals (Boston, MA), Novartis (Basel,
might just bring more choices to doctors and patients.
Switzerland), and Eli Lilly (Indianapolis, IN).
They are becoming today’s reality thanks to our
ability to isolate molecules from body fluids. These EXTRACTING EXOSOMES
diagnostic innovations pair technology with the latest
Exosomes are lipid-encased vesicles that contain cellular
in biomarkers, and are rapidly gaining acceptance as a
protein, DNA, and RNA and typically have surface
reliable way to screen for cancer and to monitor disease
proteins specific to their native cell. These attributes,
progression and response to treatment. This week we’ll
combined with the fact that they are found in many
examine the different types of liquid biopsies and how
different body fluids, make exosomes a very attractive
they work.
possibility for liquid biopsy. The idea is to capture
TERM OF THE WEEK: LIQUID BIOPSY exosomes based on tumor-specific surface markers or to
A liquid biopsy is a test that is able to detect the collect exosomes and identify them as cancer-associated
presence of cancer using blood, urine, saliva, or other by examining the enclosed DNA or RNA.
bodily fluid as the sample rather than tissue from Aptly named Exosome Diagnostics (Cambridge, MA) has
a specific organ. The technique is possible because an exosome-based urine and blood tests for prostate
cancerous tissues shed cells, DNA, and tiny lipid-encased cancer on the market, and another in development for
compartments called exosomes. Liquid biopsies detect lung cancer. Qiagen has a partnership with Exosome
the presence of these cancer-associated biomarkers. Diagnostics to help develop additional exosome-
based cancer diagnostics to complement their work
DISCOVERY BY CELL-FREE DNA with cfDNA.
When cells in the body die, they release cell-free DNA
(cfDNA)—this includes dying tumor cells. cfDNA-based CONSIDER CIRCULATING TUMOR CELLS
tests are a type of liquid biopsy because they seek out The final category of liquid biopsy is perhaps the
a biomarker – in this case, tumor DNA - in body fluids most obvious—circulating tumor cells (CTCs), or
and then identify cancer-specific mutations using PCR or cells splintered from a tumor and circulating in the
next-generation sequencing analysis. bloodstream. The challenge lies in detecting CTCs: some
Trovagene (San Diego, CA) analyzes cfDNA found in estimates classify them as rare as one circulating tumor
urine samples, which patients collect at home. Currently, cell per billion normal cells!
Trovagene has tests detecting mutations associated with Janssen Diagnostics (Raritan, NJ) currently markets
melanoma, colon cancer, and non-small cell lung cancer, CellSearch, the single FDA-approved test that allows
as well as the presence of viral DNA for the diagnosis of physicians to identify early CTCs from blood samples.
human papilloma virus. Exact Sciences Laboratories Monoclonal antibodies (mAbs) capable of recognizing
(Madison, WI) uses at-home collection in their colon proteins on the surface of migrating tumor cells are
cancer test, which analyzes cfDNA in stool samples for chemically linked to magnetic nanoparticles and then
cancer-associated DNA. added to a patient’s blood sample. These tumor-specific

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mAbs grab hold of the CTCs, and a strong magnetic field THE FUTURE
is then applied to the sample, isolating the captured cells
Ultimately, the best liquid biopsies may contain a
for identification and analysis. CellSearch is currently
combination of all the above approaches. Biocept
used to monitor the efficacy of treatments for breast,
(San Diego) is leading the way by developing liquid
prostate, and colorectal cancer. A higher number of CTCs
biopsies that analyze both cfDNA and CTCs. Biocept
detected may indicate a higher incidence of metastasis,
currently markets liquid biopsy tests for the detection
or a less than effective treatment route if used to
of lung cancer, breast, colorectal, gastric, prostate, and
quantify cancer therapy success.
melanoma. Biocept also has its eye on combination
Another way to identify CTCs may be cell size—CTCs liquid biopsies for both colon cancer and melanoma.
tend to be significantly larger than other cells in the
Today, liquid biopsies are mainly used for monitoring
blood, and this size differential may be exploited in
the progress of or response to treatment of already-
a microfluidics-based approach to cell separation.
diagnosed cancers rather than as initial diagnostic tests.
Researchers at National University in Singapore
A major goal in the field is to develop tests that can be
(Singapore) and MIT (Cambridge, MA) have developed a
used routinely to detect cancer in seemingly healthy
microfluidics chip that routes cells from a blood sample
people, which should translate to better treatment
into different channels based upon cell size. Although
outcomes. Research published by a group at Johns
still in the preclinical research phase, this approach
Hopkins University (Baltimore, MD) in January 2018
shows promise for capturing a wide range of CTCs.
suggests that a liquid biopsy test that detects both
Epic Sciences (San Diego, CA) adopts a “no cell left cancer-associated cfDNA and proteins known to be
behind” game plan thanks to technology developed by characteristic of certain types of cancer may be better at
the Scripps Research Institute (La Jolla, CA). Automated detecting cancer early on than those that look at just one
fluorescence-microscopy identifies the CTCs in blood biomarker. Dubbed Cancer-SEEK, the tool is capable of
samples placed on microscope slides. A detailed analysis detecting a number of different cancers, including ovary
of three million cells per slide is performed, each blood and liver, and may soon begin testing as a screening tool.
sample yielding approximately twelve slides. This
As the technologies to detect cfDNA, CTCs, and cancer-
technology may potentially hone in on the presence of a
specific exosomes progresses, we can expect to see
single CTC. Epic Sciences’ CTC-detecting platform is used
an increasing number liquid biopsies available, making
by Genomic Health in their Oncotype-Dx AR-V7 Nucleus
the detection and treatment of a range of cancers less
Detect test, which determines whether or not prostate
invasive and more manageable.
cancers patients’ tumors have developed mutations that
make them resistant to common types of treatment.

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TURNING ON CELLULAR GARBAGE DISPOSALS Life Science Training from Industry Experts

PROTEASOMES TO THE RESCUE “bifunctional small molecules” – which simultaneously

bind to two different proteins.
Many drugs work by stopping overactive proteins that
cause disease. The leukemia drug Gleevec, for example, With PROTAC, one end binds to the target, the other
is a small-molecule inhibitor (antagonist) of the protein to E3 ligase. This interaction transfers ubiquitin to the
Bcr-Abl, whose over-activity promotes excessive target protein for eventual disposal. PROTAC doesn’t
cell division. Humira treats a range of autoimmune necessarily have to recognize a specific part of the
diseases by stopping TNF-alpha, a protein that target, such as the active site of an enzyme. That allows
activates inflammation. researchers to focus on a wider range of proteins than
possible with existing technologies, such as small
Such antagonists can be powerful. However, it’s not
molecule inhibitors, which must fit precisely in an
always possible to develop a strong inhibitor of a
enzyme’s active site to work.
disease-associated protein. And when scientists do
develop them, resistance often emerges, rendering a Arvinas has released preclinical data suggesting that
drug ineffective. PROTAC successfully lowers levels of the protein BRD4
in lymphoma, multiple myeloma, and prostate cancer
What if, instead of merely inhibiting a protein, we
cells. BRD4 plays a role in cell division, and mutated
could totally get rid of it? It turns out that our cells
versions are associated with various cancers. In the past
already have that ability. In this Biotech Primer
year, Arvinas announced collaborations with Pfizer and
WEEKLY, we look at a new class of drugs scientists are
Genentech, which should speed the progress of getting
developing to take advantage of our bodies’ microscopic
these molecules to the clinic.
sanitation departments.
C4 Therapeutics (Cambridge, MA) is developing a
A CELLULAR GARBAGE DISPOSAL similar small molecule platform that connects disease-
associated proteins with cellular ubiquitination enzymes.
If allowed to accumulate, proteins can interfere with
Dubbed “degronimids,” the molecules are still in
normal cell function. Therefore, all cells contain
preclinical development. They have already attracted
proteasomes, compartments that break apart unneeded
the attention of Google-backed Calico, with whom C4
or damaged proteins. Proteasomal degradation also
recently signed a five-year deal to work together to treat
provides a way to recycle the amino acid building blocks
diseases of aging.
of proteins. Once a protein is broken down, a cell can use
the leftover amino acid “bits” to rebuild new proteins. Kymera (Cambridge, MA) is also utilizing small molecules
to activate target-specific proteasome degradation,
Proteins are targeted for degradation through the action
focusing first on oncology and autoimmunity. In March,
of E3 ligase. This enzyme attaches another protein,
Celgene (Summit, NJ) announced a project with Vividion
ubiquitin, to the targeted protein. Ubiquitin then guides
(La Jolla, CA) in the hopes of discovering ubiquitin-
the target into a proteasome, where it’s broken down.
proteasome system interacting drugs.
If scientists figure out how to “tag” disease-associated
proteins with ubiquitin, they can activate our cellular
NOW WAIT A SECOND…
garbage disposal to fight illness. Several companies are
working on clever ways to do just that. It’s clear that many drug developers place great faith in
tapping proteasome power to advance human health.
SMALL MOLECULES, BIG RESULTS? In what could seem completely contradictory, other
companies are taking the opposite approach: squelching
Researchers at Arvinas (New Haven, CT) are developing
the proteasome.
a platform to target disease-causing proteins based on
ubiquitination/proteasome systems. Dubbed PROTAC The process of apoptosis, or programmed cell death,
(Proteolysis-Targeting Chimera), the platform consists of occurs naturally in cells as a protective mechanism. For

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example, cells that sustain large amounts of damage to myeloma. Krypolis, developed by Onyx (South San
DNA activate apoptosis to prevent them from seeding Francisco), is another proteasome inhibitor approved as
a tumor. a second-line treatment for multiple myeloma. Both are
Many pharmaceutical companies are trying to co-opt small molecule drugs.
apoptosis to treat cancer. One way to induce the process A better understanding of how our cells process
is to inhibit the action of proteasomes. The resulting unwanted proteins has opened up an entirely new
buildup of damaged proteins signals the cell that approach to treating diseases. Manipulating the world’s
something is seriously amiss, setting off cell death. tiniest garbage disposals may hold the key to healing
The FDA has approved Velcade, marketed by Millennium otherwise untreatable conditions.
Pharmaceuticals(Cambridge, MA), to treat multiple

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HARNESSING YOUR IMMUNE
SYSTEM FOR GOOD Life Science Training from Industry Experts

YOUR INNER IMMUNE WORKINGS a bug, they engulf it – essentially eating it. Macrophages
have the ability to differentiate between good and bad
What do monoclonal antibodies, CAR-T therapy, and
bacteria and virus based on a special receptor called a
checkpoint inhibitor treatments all have in common?
PAMP (Pathogen-Associated Molecular Pattern) found
They are immunotherapies, or therapies that activate
only on the bad ones. Other non-specific defenders
the immune system to fight or prevent a disease. While
include neutrophils (which also recognize the PAMP and
an activated immune system can help save a life, an
engulf invaders) and natural killer cells, which inject the
overactive immune system can attack the body it is
protein granzyme B into invaders, triggering cell death.
charged with protecting. This over-activity is the basis for
autoimmune disorders. Once activated, these non-specific defenders release
“inflammatory cytokines,” or signaling molecules
The biotech industry has elegantly hacked the immune
that switch on other immune cells. The inflammatory
system — a highly complex network of signaling
response is kicked into gear, ensuring a rapid and
molecules, cells, and tissues — to make some of the
comprehensive retaliation.
leading immunotherapies such as Abbvie’s (North
Chicago, IL) Humira that battles psoriasis or Merck’s SPECIFIC IMMUNITY:
(Kenilworth, NJ) Keytruda that fights different cancers. T-CELLS AND B-CELLS
Let’s discover how the immune system operates and find
When non-specific defenses are unable to rid the body
out how our best and brightest are applying the immune
of pathogens, it’s time for back up. Waiting for the call
approach to disease treatment.
are T-cells and B-cells, which make up your specific (or
IMMUNE SYSTEM PRIMER adaptive) immunity. These cells are highly specialized
to recognize unique targets, called epitopes, thanks
The immune system is devoted to protecting us from
to their distinctly shaped receptors. Once the B-cell or
foreign invaders. These include viruses, bacteria,
T-cell receptor binds to the pathogen’s epitope, they
parasites, fungi, and even cancer cells. The immune
are activated. Each T- or B-cell recognizes only one
system consists of many different players, all working
unique epitope.
together as a team. The first defense consists of physical
barriers, such as the skin and mucus membranes, which
attempt to thwart these pathogens from entering our
bodies in the first place. If these barriers are breached,
then our cellular defense mechanisms kick in – first in
the form of non-specific immunity, and then in the form
of specific immunity.

NON-SPECIFIC IMMUNITY:
MACROPHAGES AND NEUTROPHILS
Non-specific (or innate) immunity fends off pathogens at
the cellular level. The troops are specialized white blood
cells (WBC). Most WBC in the body are non-specific,
meaning these foot soldiers attack in the same fashion
without stopping to consider the specific characteristics
Activated T-cells divide rapidly and produce three types
of the enemy.
of descendants: killer T-cells, helper T-cells, and memory
Macrophages are one type of non-specific defender that T-cells. All recognize the same target as the originally
freely circulate in the bloodstream. When they encounter activated T-cell.

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 • Killer T-cells roam the body in search of their pre- antibody Herceptin, binds to the HER2 epitope which is
programmed epitope, and seal the deal by injecting present at high levels on the surface of 25% of breast
granzyme B, triggering cell death. cancer patient’s tumors. This compels white blood
• Don’t let the label “helper” fool you – helper T-cells cells, such as killer T-cells and macrophages, to attack
are critical, they release inflammatory cytokines that the tumor. Herceptin acts just like a naturally-occurring
activate antibody-producing B-cells, killer T-cells, antibody, only it is produced in the lab. Chimeric antigen
and macrophages to respond en masse. The human receptor T-cell (CAR-T) therapies, which to date are still
immunodeficiency virus (HIV) only infects helper in clinical testing, are T-cells whose receptors have been
T-cells, and in so doing completely cripples the engineered to recognize and destroy cancer.
immune response.
CHECKPOINTS ON THE CASE
• Memory T-cells don’t defend against an initial
The body has natural checkpoints to prevent
infection, but if these cells encounter the same
inflammatory disorders; this screening process stops
epitope a second time, they are very quickly
T-cells and B-cells from mistakenly killing its own tissues.
converted into killer T-cells and helper T-cells,
These checkpoint proteins send a “stop attacking”
ensuring a rapid response.
signal to the T- and B-cell when they encounter their
body’s healthy cells. Many types of cancer cells have
evolved to express these checkpoint proteins, tricking
the T and B-cell into thinking the cancer is a healthy
cell. Checkpoint inhibitor therapies prevent the cancer
from activating their checkpoint proteins, enabling the
immune system to more fully go after the tumor.

THE OTHER SIDE OF THE COIN:

AUTOIMMUNE DISEASE
The immune system prevents us from falling deathly ill
as it responds to constant microbe exposure. However,
an overactive immune system can cause serious
problems, potentially leading to autoimmune disease.
Chronic inflammatory disorders such as Crohn’s disease,
Activated B-cells also reproduce rapidly and produce
rheumatoid arthritis, and psoriasis wreak havoc by
two types of descendants: plasma cells and memory
activating white blood cells to target innocent cells in
B-cells.
the body and release inflammatory cytokines to sustain
• Plasma cells secrete antibodies - proteins that the barrage.
recognize and bind to any bacterium or virally-
Biologic drugs that treat these disorders—like Humira
infected cell that bears a matching epitope. The
(AbbVie, North Chicago, IL), Enbrel (Amgen, Thousand
binding action triggers other immune cells, such
Oaks, CA), and Rituxan (Genentech, South San Francisco,
as killer T-cells or macrophages, to sweep in and
CA)—work by shutting down key parts of the response.
destroy the invader attached to the antibody.
Humira and Enbrel inhibit a specific inflammatory
• Like memory T-cells, memory B-cells lie in wait, cytokine known as TNF-alpha. Both of these drugs are
preparing for future attacks instigated by the same approved for a range of inflammatory diseases. Rituxan,
foreign invader. approved for rheumatoid arthritis, works by reducing the
number of B-cells that target the synovial tissue of joints.
APPLIED IMMUNOLOGY
From one side of the coin to the other, the immune
Immunotherapies use strategies from specific immunity system continues to both challenge and reward the
to their advantage. Monoclonal antibody therapies are industry as new pathways and targets are discovered.
developed by selecting antibodies that recognize and A delicate balance of the body’s toughest fighters,
bind to a disease-specific epitope. The classic example, understanding and optimizing the immune system is
Genentech’s (South San Francisco, CA) therapeutic central to the immunotherapy paradigm.

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DECODING THE GUT-BRAIN AXIS Life Science Training from Industry Experts

MICROBIOME & DISEASE diseases such as obesity, diabetes, and, increasingly,

brain disorders.
The biotech world is abuzz with talk of the latest
microbiome startup launch Axial Biotherapeutics TRAIL BLAZERS: AXIAL
(Boston, MA). What caught our eye here at the WEEKLY BIOTHERAPEUTICS & THE
is the company’s focus on the gut microbiome — the MAZMANIAN LAB
entire collection of microbes living in the gut — and its
Axial is focusing much of its initial attention on work
interplay with the brain.
originally done at microbiologist Sarkis Mazmanian’s
The connection between the gut microbiome and lab in the California Institute of Technology (Pasadena,
inflammatory bowel disease is much more intuitive CA) — mostly relating to the gut-brain axis’ role in
since they occur in the intestine. In the past few years, Parkinson’s disease (PD) and autism.
however, new research has made it clear the gut
microbiome also impacts neurological health, leading Parkinson’s Disease
to the phrase “the gut-brain axis.” Let’s explore this Parkinson’s disease (PD) is a chronic and progressive
connection and examine the early efforts to translate movement disorder, according to the Parkinson’s
these discoveries in the clinic. Disease Foundation. Symptoms include tremor of
the hands, arms, legs, jaw, and face; slowness of
GUT MICROBIOME PRIMER movement; rigidity of the limbs and trunk; and impaired
The human microbiome is the complex collection of balance and coordination. These symptoms are
microbes (mostly bacteria, but also includes small caused by the malfunction and death of neurons that
numbers of fungi and viruses) that reside on and produce the neurotransmitter dopamine. PD affects
inside the human body, including our skin, mouth, nearly one million people in the U.S., and the cause is
nose, respiratory tract, and digestive tract (gut). The unknown. About 75% of PD cases are accompanied by
microbiome is huge — microbial cells outnumber human gastrointestinal disorders such as constipation, which
cells by a ten to one ratio! Human cells are much larger provided an initial impetus to examine a possible
than bacteria cells, however, so don’t worry — you’re still connection between gut health and the disease.
mostly human. For every 100 pounds that you weigh, it A key molecular characteristic of PD is the aggregation
is estimated that about two pounds of that weight come of a protein called alpha-synuclein (αSyn) within cells of
from bacteria. the brain and gut. Researchers in the Mazmanian lab
Most of us think of bacteria as harmful and certainly used a strain of mice that over produce αSyn to study the
some types are; however, those that have evolved with disease. One group of mice were bred in a completely
humans to become part of the human microbiome sterile environment to create “germ-free” (GF) αSyn mice.
are either neutral — causing no harm — or beneficial. The other αSyn mice had the normal collection of gut
Scientists are busy trying to better understand and bacteria. On a series of tests designed to assess motor
characterize these beneficial bacteria and the role that skills, the GF αSyn mice performed significantly better —
they play in human health. suggesting that even in mice that overproduced the αSyn
The gut contains the largest number of bacteria, protein, the presence of certain microbes are required
as well as the greatest diversity of bacteria, when for the disease to progress. Further work suggested
compared to other parts of the body. Thus much of that a molecule called butyrate, produced by certain gut
the attention directed towards the human microbiome bacteria, can enter the brain and activate an immune
has been focused on the gut microbiome in particular, response, leading neurons to malfunction or die.
which continues to surprise us with its influence on There is reason to believe that this connection is also
at work in humans with PD. In collaboration with Rush

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 These symptoms impact a child’s educational, social,
emotional, and physical development. More than 3.5
million Americans live with an autism spectrum disorder
according to the Autism Society. The cause is unknown,
although genetics is thought to play a role.
Similar to PD, a significant portion of ASD patients
exhibit gastrointestinal problems. The Mazmanian
lab demonstrated that feeding ASD mice a specific
strain of bacteria called B. fragilis — a part of the
human microbiome — altered the mouse microbiome
and reduced some of the ASD-like behaviors such as
anxiety and repetitive behavior, and increased levels
of communication with other mice. These experiments
suggest a possible probiotic therapy for autism.

CARB LOADING WITH SYMBIOTIX

Another early-stage company making headlines in the
gut-brain axis space is Symbiotix (Boston, MA). Focusing
on multiple sclerosis (MS), their lead candidate is a
carbohydrate molecule produced by B. fragilis. This
University (Chicago, IL) gastroenterologists, Mazmanian therapy increases the production of regulatory T-cells,
lab researchers transferred fecal samples from PD which are a class of T-cells that “turn down” an overactive
patients into the GF αSyn mice. Fecal transplants are immune response by releasing anti-inflammatory
an established way to “reset” the gut microbiome of signaling molecules. Symbiotix is preparing to enter
the recipient to make it match that of the donor. After clinical trials with an orally-administered product for the
transplantation, the mice began exhibiting symptoms of treatment of MS and inflammatory bowel disease.
PD. Transfer of fecal matter from healthy people did not
trigger these symptoms. These experiments suggest that KALLYOPE ON THE LOOKOUT
the gut microbiome is a major contributor to the disease
Launched just one year ago, Kallyope (New York City,
process in PD patients.
NY) also hopes to translate our growing knowledge of
Of course, these promising early stage findings still need the gut-brain axis into new therapeutics. The company
to be translated into human therapeutics. This may be is developing a discovery platform using tools such
easier than traditional neurological approaches because as genome sequencing and brain imaging to precisely
it is much easier to deliver drugs to the gut than to get identify gut-microbiome-derived molecules of potential
them to cross the blood-brain barrier. Following up with therapeutic interest.
a targeted approach to modulate the production of
The emerging work described here gives credence to the
butyrate and other inflammatory compounds produced
old expression “think with your gut.” In addition to the
in the gut may bring the first truly effective PD therapy
diseases discussed above, scientists are discovering links
into the clinics.
between the gut microbiome and other brain disorders
Autism Spectrum Disorder such as anxiety and depression. As the story continues
to unfold, we are likely to see new therapeutics based on
Autism spectrum disorder (ASD) is a developmental brain
restoring the balance that millions of years of human-
disorder characterized by impaired social interaction,
microbe co-evolution has fine-tuned.
communication, and restrictive and repetitive behavior.

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UNWIRING THE BIOLOGY OF FIBROSIS Life Science Training from Industry Experts

FIBROTIC DISEASE INNOVATIONS INTERCEPTING IPF

Fibrotic diseases — organ and tissue disorders that Several different companies are going after idiopathic
occur as a result of the buildup of excessive scar pulmonary fibrosis, or fibrosis of the lung whose exact
tissue — are sounding the alarm in the biotech sector. origin is not known. IPF is more common in smokers, and
A silent epidemic creeping up on the Western world recent studies report that 9/11 first responders suffered
involves non-alcoholic steatohepatitis (NASH), or liver from increased rates of IPF, presumably due to breathing
fibrosis associated with obesity and type 2 diabetes. in high levels of toxic smoke at the site. This suggests a
Left untreated, NASH can lead to liver failure. Currently, possible link between IPF and tissue damage caused by
NASH population estimates run in the tens of millions smoke inhalation, at least in some cases.
of people in the U.S., and the numbers are expected Currently, there are two FDA-approved drugs on the
to grow along with the incidence of obesity and type market for IPF:
2 diabetes. In the same vein, idiopathic pulmonary
• Ofev (Boehringer Ingelheim; Ridgefield, CT) is
fibrosis (IPF) is another type of fibrotic disease affecting
a small molecule inhibitor of signaling pathways
approximately 200,000 a people per year in the U.S.
thought to be involved in lung tissue scarring.
The severity of IPF manifests itself as progressive
breathlessness leading to respiratory failure, resulting in • Esbriet (Roche; Basel, Switzerland) is a small
40,000 deaths per year. molecule drug that reduces fibroblast proliferation
and collagen production, and reduces the production
In this WEEKLY, we will delve more deeply into the roots
of inflammatory signaling molecules. The exact
of fibrosis and break down some of the latest treatment
mechanism by which Esbriet exerts these effects is
innovations in the clinic.
not known.
TERM OF THE WEEK: FIBROSIS Pliant Therapeutics (Redwood City, CA) is developing
Connective tissue forms a wiry network which provides a small molecule inhibitor of protein receptors known
the support and structure to cells making up an organ or as integrins. Integrins play a key role in activating the
tissue. It consists of cells such as fat cells or a special cell collagen-producing fibroblast cells. By inhibiting a subset
type called fibroblasts which are suspended in a gel-like of these integrins present on the surface of the lung,
matrix made up of proteins. The key protein component Pliant hopes to prevent the growth of fibrotic tissue
of this matrix is collagen, a protein that takes the form of within the lung, since scar tissue formation is associated
elongated, fine fibers, providing flexible support to the with excess collagen production. Initial clinical trials for
surrounding cells. IPF are slated in 2017, with plans for subsequent trials in
liver (NASH) and kidney fibrosis.
Fibrosis comes into the picture when the threadlike
connective tissue becomes excessive. This can be Biogen (Cambridge, MA) has a monoclonal antibody
triggered by injury and the resulting inflammation. In therapeutic, STX-100, in Phase II clinical studies for IPF.
an attempt to repair damaged tissue, activated immune The drug works by inhibiting the inflammatory signaling
cells release factors that stimulate fibroblasts to secrete molecule TGF-beta, which is a key driver of fibrosis.
extra collagen and other proteins. This process is
normally very carefully controlled, but repeated injury or
persistent inflammation can dysregulate it, resulting in
scarring, thickening, and loss of function in areas such as
the liver, lungs and more.

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CONFRONTING NASH
There are no drugs approved specifically for the
treatment of NASH in the current market. Since the scar
tissue present in NASH is caused by inflammation of the
liver associated with the excessive accumulation of liver
fat, many of the treatments in development are focused
on managing lipid metabolism, glucose regulation,
and inflammation.
Intercept Pharmaceuticals (San Diego, CA) has a
promising drug in Phase III clinical trials for NASH. The
small molecule drug, obeticholic acid, works by binding
to a so-called nuclear receptor – a receptor protein that
is present inside of cells rather than on their surface.
When this internal receptor is activated, it moves into the
cell’s nucleus, where it binds DNA at a specific location, Another recent acquisition in the NASH field is
activating the expression of particular genes. Obeticholic Allergan’s (Parsippany-Troy Hills, NJ) purchase of
acid binds to and activates the nuclear receptor FXR. Tobira’s (South San Francisco, CA) cenicriviroc, a small
FXR then modulates the expression of a variety of genes molecule inhibitor of receptors present on the surface
involved in lipid metabolism and glucose homeostasis. of macrophages, a type of white blood cell important in
Obeticholic acid has been granted breakthrough therapy the inflammatory response. When these receptors are
status by the FDA. triggered, the macrophages become fully activated and
promote the inflammatory processes that contribute to
Bristol-Myers Squibb (New York, NY) ) recently acquired
NASH. Cenicriviroc is in Phase I clinical development.
a promising new NASH candidate, ND-L02-s0201, from
the Japanese biotech Nitto (Osaka, Japan). The drug is an Gilead (Foster City, CA) is also pursuing an anti-
antisense drug, which means it works by preventing the inflammatory approach in its small molecule NASH
production of a specific protein. ND-L02-s0201 blocks treatment Selonsertib, which works by inhibiting ASK1,
a protein called “heat shock protein 47 (HSP47),” which an enzyme that activates signaling pathways involved in
is required for the production of functional collagen. inflammation, cell death, and fibrosis.
So by blocking the production of HSP47, the production With increasing awareness and attention on fibrotic
of scar-tissue promoting collagen should decrease. disease, we can expect to see significant progress in this
ND-L02-s0201 has recently completed Phase Ib/II arena within the next few years.
clinical studies.

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THE SCIENCE BEHIND THE OPIOID EPIDEMIC Life Science Training from Industry Experts

A BIG PAIN medications, such as Purdue Pharma’s (Stamford,

CT) Oxycontin and Endo Pharmaceuticals’ (Malvern,
The opioid addiction crisis gained attention at the
PA) Percocet, work by binding to mu receptor proteins
highest levels of U.S. policy circles this past year.
on the surface of cells in the central nervous system
Presidential candidates that disagreed on nearly
(CNS) — like the brain and spinal cord. While the CNS
everything else vowed to make fighting the epidemic a
is tasked with relaying pain signals, opioids decrease
priority if elected.
the excitability of nerve cells delivering the message,
In July, the U.S. Senate overwhelmingly approved a resulting in pain relief — along with a feeling of euphoria
bill to strengthen prevention, treatment, and recovery in some users.
efforts. And no wonder – according to the Centers for
Disease Control, opioid overdose deaths are at an all- RELIEVING THE PAIN
time high, a stark reality that highlights the dark side of According to the Centers for Disease Control, 44
a class of treatments serving a vital need. Opioid pain Americans die every day due to prescription painkiller
medications manage the severe short-term or chronic overdose. At the same time, chronic pain is also a serious
pain of millions of Americans. problem, affecting approximately 100 million U.S. adults,
While these medications mitigate needless suffering, while millions of others suffer acute pain due to injury
joining forces are the government, corporations, and or surgery. The medical need for these drugs is very real
medical community to battle against opioid abuse and despite the dark side.
addiction. Here at the WEEKLY, we always wonder: Short term medical use of opioid pain killers rarely leads
what is the science behind the story? So, let’s talk about to addiction when properly managed. The euphoria-
how pain medications work, the different types on the inducing effects of the drugs, long-term regular use,
market, and the approaches to developing less addictive or use in the absence of pain, can lead to physical
versions of opioid drugs. dependence and addiction. And because regular use
increases drug tolerance, higher doses are required to
OPIOIDS VS. NSAIDS achieve the same effect, leading abusers to consume
There are two main categories of pain medications, pain pills in unsafe ways such as crushing and snorting
opioids and non-steroidal anti-inflammatory drugs or injecting the pills.
(NSAIDs). Although these two categories of drugs work
The answer to developing less addictive drugs may
differently, they do share one thing in common: both are
be found in a drug that blocks pain without inducing
derivatives of natural products.
euphoria. These new drugs will need a different
The NSAID Aspirin (Bayer, Leverkusen, Germany) mechanism of action than traditional opioid drugs, which
is a synthetic version of an extract from willow tree bind to the mu receptors of cells inside the CNS.
bark. Aspirin works by inhibiting an enzyme called
Cara Therapeutics (Shelton, CT) is developing drugs
cyclooxygenase 1 (COX-1). Once stopped, COX-1 is no
that bind to a different type of opioid receptor, the
longer able to produce signaling molecules, called
kappa opioid receptor. These receptors are present on
prostaglandins and thromboxanes. Prostaglandins and
sensory nerves outside of the CNS. Preclinical studies
thromboxanes have a wide variety of functions, including
suggest that targeting these receptors could be effective
mediating aspects of inflammation (fever and swelling)
at reducing pain without driving addictive behaviors.
as well as promoting neuronal response to pain. Other
Their lead candidate, CR845, is currently in Phase III
NSAIDs, such as ibuprofen and naproxen, also work by
clinical testing for post-operative pain and pruritus
inhibiting COX-1 or its sister enzyme COX-2.
(severe itching), and in Phase II clinical testing for chronic
Opioids are synthetic versions of opium and morphine, pain. Cara Therapeutics is also developing compounds
which come from poppy flowers. Opioid pain that selectively activate cannabinoid (CB) receptors

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outside of the CNS. CB receptors inside the CNS are by the body that bind to and activate the mu opioid
linked to the psychoactive qualities of marijuana; those receptors, resulting in pain relief and mild euphoria
outside the brain are found on white blood cells and without the detrimental side effects associated with
have been shown to be involved in decreasing pain and opioid drugs such depressed respiration, motor
inflammation. Cara’s lead CB receptor activator, CR701, is impairment, and addiction. Scientists have tried before
in preclinical development. to develop safer pain medications based on endogenous
Hydra Biosciences (Cambridge, MA) is developing small opioids, but have not been successful due to the
molecule inhibitors of ion channels – proteins on the instability of these molecules. The Tulane team created a
surface of nerve cells that help to transmit pain signals derivative of endomorphin that is stable and binds to the
by allowing positively charged calcium ions to enter the mu receptor in such a way that pain relief occurs, but not
nerve. This plays a critical role in sending the pain signal the negative side effects listed above. Clinical testing is
to the brain, yet because it works on nerves outside of expected to begin by the end of 2017.
the brain, it has less of a potential for addiction. Hydra
is currently in Phase I clinical studies of HX-100 for the
AN ANTIDOTE TO AN OVERDOSE
treatment of painful diabetic neuropathy. Overdosing can be fatal since respiratory failure occurs
at high blood concentration levels of opioids. If an
Centrexion Therapeutics (Baltimore, MD) lead
overdose is suspected, the individual should be treated
candidate is a derivative of capsaicin, a naturally-
as quickly as possible with naloxone — a “competitive
occurring compound found in chili peppers. Capsaicin
antagonist” of the mu opioid receptor. Simply put, a
has pain relieving properties and has been used as a
competitive antagonist binds the receptor without
natural remedy. Centrexion’s lead candidate, CNTX-4975,
activating it. Since naloxone doesn’t activate the
is a highly potent, synthetic form of capsaicin designed
receptor, it doesn’t have any pain-relieving or euphoria-
to be administered via injection into the site of pain.
inducing qualities; rather, it prevents the opioid drugs
CNTX-4975 targets the capsaicin receptor, an ion channel
from binding. It may also displace opioids that have
protein on the surface of nerve cells. When CNTX-4975
already bound the mu receptor, aiding in the stoppage of
binds the capsaicin receptor, the influx of calcium ions
an overdose.
results in desensitization of the nerves, making them
unresponsive to other pain signals. This effect can last
for months, and only affects nerves near the site of
COCKTAIL FODDER: RUNNER’S HIGH
injection. CNTX-4975 is currently in Phase IIb clinical Some folks love to run; others avoid it at all costs. This
studies for knee osteoarthritis, and Phase II clinical might be explained by inherent differences in sensitivity
studies for Morton’s neuroma, a sharp pain in the foot to the natural opioids called endorphins that are
and toe caused from a thickening of the tissue around released during exercise. Not everyone experiences the
one of the nerves leading to the toes. “runner’s high”— feelings of calm and mild euphoria
– just like not everyone experiences euphoric feelings
Earlier this year, researchers at Tulane University
from pain medications. These differences may help to
published a paper that shows great promise for the
explain why some people enjoy exercise and others
development of effective, yet non-addictive pain
don’t, and why some people get addicted to opioids—
medications. They have developed a compound that
while others can take them or leave them.
is derived from the endogenous opioid endomorphin.
Endogenous opioids are chemicals produced naturally

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THE MOLECULAR CAUSE OF OBESITY Life Science Training from Industry Experts

TARGETING FAT difficult and most patients regain any lost weight within
two years.
Obesity is considered one of the most pressing public
health issues of the day. According to the Center for Obesity is associated with increased risk for other
Disease Control, 37% of adults and 17% of children in metabolic, cardiovascular, and inflammatory disorders
the U.S. are obese. such as non-alcoholic fatty liver disease, hypertension,
coronary heart disease, and stroke. It is also the primary
The latest drug interventions work by attempting to
cause of type 2 diabetes.
suppress food intake, which has proven beneficial for
some. However, there is still a large unmet need for new A VICIOUS CYCLE
therapeutics that target the underlying molecular causes
Fat used to be thought of as simply a way to store
of obesity — starting with adipose (fat) tissue.
energy. Excess calories get converted into fat, which
In this WEEKLY, we’ll explore the emerging view that can then be broken down into fatty acids, serving as
adipose tissue is a metabolic organ undergoing an energy source when needed. Interestingly, the
pathological changes during obesity progression and total number of the cells making up adipose tissue —
find out which drugs are being developed to fight adipocytes — remains the same once a person reaches
this epidemic. adulthood. When someone gains weight they don’t
increase the number of adipocytes, they increase the
TERM OF THE WEEK: OBESITY actual size of adipocytes. Researchers have found that
Obesity is defined as a medical condition in which excess location of that weight gain matters: increased adipose
body fat has accumulated to the extent that it is likely to tissue within the abdomen is linked to metabolic disease;
have a negative effect on health. Clinically, a person is subcutaneous — just beneath the skin — adipose
usually considered obese when their body mass index — accumulation is not.
a number obtained by dividing a person’s weight by the
A number of things happen as abdominal adipocytes
square of their height — is over 30 kg/m2.
get bigger. The types of “adipokines” — metabolic
Obesity is typically caused by a combination of lifestyle signaling molecules that enable communication between
and genetics, although there are a few forms that are adipose tissue, brain, liver, immune system, and other
driven entirely by genetics. Lifestyle interventions that organs — change. An increase in fat causes adipokines
alter diet and increase exercise are successful in some; to move from molecules that suppress inflammation to
however, long-term maintenance has proven to be those that promote inflammation. This, in turn, attracts
macrophages to the tissue. Macrophages are a type of
white blood cell that further promote inflammation,
resulting in decreased insulin sensitivity throughout the
body. As adipocytes grow big, they outpace the ability
of the blood vessel networks to support their growth.
This results in low tissue oxygen levels, which further
promotes the production of inflammatory adipokines,
again contributing to insulin insensitivity. Ultimately, the
abdominal adipocytes become so dysfunctional that
they can no longer increase their lipid deposits, so fat
expansion begins to occur in the liver, skeletal muscle,
and pancreas — disrupting their functions, including
insulin production (pancreas) and response (liver,
Brown Adipose White Adipose skeletal muscle).

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This deeper understanding of adipose tissue, as a conditions, both are currently in Phase I clinical
result of obesity, has led to some new drug discovery studies for obesity and type 2 diabetes.
and development approaches. The therapeutic goal: • IL-1 beta inhibitors: A second key pro-inflammatory
to reduce some of the systemic effects such as insulin signaling molecule is IL-1 beta. Amgen (Thousand
resistance and inflammation. Oaks, CA) is conducting preclinical trials for obesity
and type 2 diabetes of their IL-1 beta inhibitor
AIMING FOR ADIPOKINES Anakinra, which is already approved for rheumatoid
One of the beneficial adipokines disrupted in obesity is arthritis. Novartis (Basel, Switzerland) is doing the
adiponectin. Adiponectin increases insulin sensitivity same for their drug Ilaris, which is approved for a
and decreases inflammation. Researchers at the range of different inflammatory conditions.
University of Tokyo are conducting preclinical testing of
• Blocking macrophage movement into adipose tissue:
AdipoRon, a small molecule activator of the adiponectin
Macrophages move into adipose tissue in response
receptor present on the surface of muscle and liver
to signaling molecules called “chemokines.” By
cells. Scientists have demonstrated AdipoRon’s ability
inhibiting chemokine signaling, researchers think
to reduce insulin resistance, abnormal distribution of
they may be able to block macrophages from
adipocytes, and glucose intolerance in mice that have
entering adipose tissue and worsening the cycle of
been bred to be a model for obesity and type 2 diabetes.
inflammation. Chemocentryx (Mountain View, CA),
The most famous adipokine is leptin, a hormone that Merck (Kenilworth, NJ), and Bristol-Myers Squibb
promotes feelings of satiety. Myalept is a leptin analog (New York City, NY) currently have drugs in Phase II
that is already approved for the treatment of the rare clinical studies that do just that.
disorder lipodystrophy, or the irregular distribution
of adipose tissue within the body. Myalept is currently WHITE VS. BROWN ADIPOSE TISSUE
in Phase II clinical testing for obesity by Aegerion White adipose tissue (the type described above) is
Pharmaceuticals (Cambridge, MA). associated with obesity and insulin resistance. Brown
adipose tissue, in contrast, regulates body temperature
MITIGATING INFLAMMATION by actually burning calories to release heat and appears
Since inflammation appears to be a root cause to have a beneficial metabolic impact. In most adults,
of adipose tissue dysfunction, the use of anti- however, brown adipose makes up only ~5% of total
inflammatories is an attractive therapeutic option. A adipose tissue. Scientists are also investigating the
number of different companies are testing approved or potential for making white adipose tissue behave more
newly developed anti-inflammatory therapeutics: like brown adipose tissue.
• TNF inhibitors: One of the key drivers of inflammation Taken together, these various approaches should yield
is the signaling molecule TNF. Remicade ( Janssen more effective treatments for obesity and insulin
Biotech, Horsham, PA) and Enbrel (Pfizer, New York resistance in the coming years.
City, NY) both work by inhibiting TNF. Though already
approved for a range of different inflammatory

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CANCER VACCINES & GAME CHANGERS Life Science Training from Industry Experts

THE ELUSIVE CANCER VACCINE FINDING NEOANTIGENS

The promise of cancer vaccines have proven to be One of the hallmarks of cancer cell development is a high
elusive. A new crop of biotechs are hoping to change rate of DNA mutation. Once a tumor is established, it
that by taking advantage of the latest advances in may grow to have dozens or even hundreds of mutations
genomics. Scientists are working overtime trying to that differentiate it from healthy cells. Identifying these
develop cancer vaccines that train the immune system cancer mutations begins with performing a biopsy
to recognize and fight an established tumor. In this to collect and study a small sample of tumor cells.
WEEKLY, we’ll break down the science and technology of Thanks to advances in genome sequencing technology,
immunotherapeutic vaccines. researchers can sequence the tumors entire “exome”—
the portion of the DNA used to make proteins. Since only
TERM OF THE WEEK: NEOANTIGEN the exome is used to produce antigens, scientists look
An antigen is a protein or portion of a protein present in only at the exome to identify neoantigens. Tumor cell
a cell that is recognized by the immune system. Think of exomes are then compared to healthy cell exomes, and
antigens as flags; some flags are “good” and some flags the differences in DNA sequences are identified.
are “bad.” An immune response occurs when attack cells, It turns out not all proteins make good antigens, the
such as macrophages and cytotoxic T-cells, encounter best are those displayed on the cell surface where they
a “bad” flag. The best case scenario: cells waving the can be recognized by the immune system. In order
“bad” flag or antigen are recognized, targeted and killed. to identify cell surface neoantigens, the “unique to
Worst case scenario: cells waving the “bad” antigen cancer DNA sequences” are fed into bioinformatics
have evolved strategies to operate in stealth mode. programs which predict the probability of surface
This cloaking mechanism is often used by cancer — it is location. Typically around 5% of the mutated genes are
the reason why it can be a silent disease until the very potential neoantigens.
late stages.
As a tumor grows, it can accumulate additional TRAINING THE VACCINE
mutations (changes in the DNA). Scientists have found Once neoantigens are identified, they are synthesized
some of these mutations are significant enough to in the lab and mixed with an adjuvant — a substance
produce new antigens which the immune system can that boosts the overall immune response. Ideally, a
recognize. These are called neoantigens, and they are neoantigen-based vaccine contains at least twenty
the secret sauce in cancer vaccines. different neoantigens, both to produce a strong
immune response and to reduce the likelihood of
resistance. A tumor may mutate and stop producing
one neoantigen, but it is unlikely to stop producing
several simultaneously.
Although in some cases, several different patients may
share common neoantigens, others may be unique to
a given patient. So it is likely that these types of cancer
vaccines would be truly personalized medicine —
designed just for one patient. This type of precision was
unimaginable even just a few years ago, but is possible
today because of the increased efficiencies in both time
and money achieved with genome sequencing.

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THE GAME CHANGERS combination with Bristol-Myers Squibb’s (New
York, NY) already approved checkpoint inhibitor
Several neoantigen-based vaccines are already in clinical
therapy Opdivo.
testing. Currently, the time taken to identify neoantigens
and produce a vaccine is around six to twelve weeks; • Gritstone Oncology (Emeryville, CA) and Immune
the goal is to bring this development time down to one Design (Seattle, WA) have a partnership to discover
month. Key players include: and develop neoantigen-based vaccines, with
clinical trials for a non-small cell lung cancer product
• TapImmune ( Jacksonville, FL) began Phase II clinical
expected to begin in 2017. Patients in the trial will
studies for a neoantigen-based vaccine (TPIV
be treated with a neoantigen vaccine in combination
200) for triple-negative breast cancer, the most
with an as yet unnamed immune-checkpoint
difficult type of breast cancer to treat using today’s
inhibitor. Checkpoint inhibitor treatments relieve
therapies. TPIV 200 will be tested in combination
the natural inhibitions on cytotoxic T-cells, enabling
with AstraZeneca’s (London, UK) durvalumab,
the immune system to become fully activated in
a checkpoint inhibitor currently in Phase III
response to the vaccine.
clinical testing.
If any of these companies succeed, we will be witness to
• Neon Therapeutics (Cambridge, MA) is testing
truly personalized medicine at work.
its melanoma and glioblastoma neoantigen
vaccine, NEO-PV-01, in Phase Ib clinical studies in

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THE RACE TO BEAT SMA Life Science Training from Industry Experts

ZEROING IN ON THE SMA PIPELINE produced. It has been found that patients with less
severe forms of the disease usually have extra copies
A decade ago, there was only one drug in development
of SMN2. This shows that even if small amounts of
for a debilitating neuromuscular disorder known as
functional SMN protein are made from each SMN2 gene,
spinal muscular atrophy (SMA) — robbing people of their
the extra copies of the SMN2 gene ultimately translate
ability to walk, eat, and ultimately, breathe.
into more functional protein overall, which helps to
Today, there are 14 therapies making their way through restore at least some motor nerve function.
the clinic according to the patient advocacy group
The four generally accepted classifications of SMA are:
Cure SMA. The increase is largely due to a better
understanding of the disease, more awareness, and • Type 1: The most severe and the most common type
a surge in funding for basic and clinical research. of SMA. Symptoms are usually present within the
SMA affects about 1 in 10,000 babies born in the first few months of life, and these babies often do
United States. not display movement of any kind. As the disease
progresses, toddlers have trouble with swallowing
In this weekly, we’ll decipher the science behind SMA
and respiratory function. SMA Type 1 is usually fatal
and find out how developmental drugs are zeroing in on
by age two.
ways to conquer this genetic disease.
• Type 2: Symptoms manifest between six and
SMA PRIMER eighteen months. These children can typically sit
but not stand or walk. Respiratory function is often
Our motor neurons work to pass messages throughout
compromised and is a major concern, however with
the body via the nervous system. In order for the motor
the help of machines many of these patients live
neurons outside of our brain to do their job, a protein
into adulthood.
called the survival motor neural (SMN) protein is critical
to keeping the motor neurons functioning and healthy. • Type 3: Symptoms occur after age one. These
The survival motor neuron 1 (SMN1) gene is the gene patients are usually able to walk, but may lose
responsible for producing most of the survival motor that ability as the disease progresses. Respiratory
neural protein used by the body. A second, closely function is less impaired, and life expectancy is often
related gene is the survival motor neuron 2 (SMN2) gene, near normal.
which produces a much smaller amount of SMN and is • Type 4: This is the adult-onset form, typically
seen as a sort of “back-up” version of SMN1. manifesting at age 30 or later. Muscles gradually
SMA is caused by a variety of mutations in the SMN1 weaken, and the patient often needs to use
gene. Without the functional SMN, the neurons do a wheelchair later in life. Life expectancy is
not work correctly and eventually die. How soon they not affected.
die depends on the extent of the SMN deficit, which Because SMA Type 1 is the most common and severe –
correlates with the severity of the disease: the less SMN about 60% of cases – most of the drugs in development
produced, the more severe the disease. aim to tackle this portion of the disease population.
The back-up gene, SMN2, produces a small amount of Successful therapies will likely later be tested in the less
functional SMN protein; however, differences in the severe forms of SMA. Below we list some approaches
way this gene functions means most of the protein going after the root cause of this orphan disease—not
is nonfunctional and degrades shortly after being enough SMN1 protein production.

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MAKING SENSE FROM ANTISENSE GENE THERAPY CURE?
The most advanced drug in the SMA pipeline is As a single gene disorder, SMA is an ideal candidate for
Nusinersen. Ionis Pharmaceuticals (Carlsbad, CA) gene therapy approaches because delivering a “good”
partnered with Biogen (Cambridge, MA) on the antisense copy of the mutated gene could potentially fill in the
therapy, and clinical trial data supports its efficacy in blanks to get the SMN1 gene working.
SMA Type 1 patients. With Phase III trials complete, a Using a “vector” — a virus stripped of its disease-causing
New Drug Application (NDA) has been filed. If the NDA ability — scientists are able to safely deliver corrected
is approved by the FDA, Nusinersen will be given the go genes into targeted cells. In the case of SMA type 1, the
ahead to be marketed in the USA. AAV9 vector crosses the blood-brain barrier and delivers
Nusinersen is an antisense drug, meaning it is synthetic corrected copies of the SMN1 gene into motor neuron
mRNA. Nusinersen (synthetic mRNA) binds to the cells in the brain.
naturally occurring SMN2 mRNA in such a way that AveXis (Bannockburn, IL) has a gene therapy
more of the mRNA is used to make protein—resulting candidate, AVXS-101, in Phase I/II clinical studies.
in greater amount of full-length, functional SMN The company reported promising results over the
protein. Recall from high school biology that mRNA is summer, noting that the drug appears to be safe and
the instruction of how to make a protein. If the mRNA is effective. Preliminary observations show babies who
working properly, the correct, functional protein is made. received this gene therapy showed marked increases
in SMN production and in movement. Also in the mix
SMALL MOLECULE ENHANCERS is Voyager Therapeutics (Cambridge, MA) with their
PTC Therapeutics (South Plainfield, NJ) — in partnership gene therapy treatment for SMA Type 1, currently in
with Roche (Basel, Switzerland) — has begun Phase II preclinical development.
clinical studies on its proprietary small molecule drug,
As these drugs move through clinical trials, SMA patients
RG67800. RG67800 is similar to Nusinersen in that it
and their family members are putting their trust into
changes the way nerve cells process the SMN2 mRNA,
these new therapies and hoping to one day kiss this
resulting in increased production of functional SMN
devastating illness good-bye.
protein. However, there is a notable difference is the
delivery mechanism — Nusinersin is an injectable while
RG67800 is a pill. Novartis (Basel, Switzerland) also has
a small molecule modulator of SMN2 mRNA in Phase II
clinical studies.

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KILLING CANCER AT ITS STEM CELL ORIGIN Life Science Training from Industry Experts

KILLING CANCER tumor recedes, the CSCs remain a possible source of

reemergence, pointing to the need for therapies that
When it comes to killing cancer, eradicating every
exterminate cancer at its source.
single cell is the goal. But did you know there are
different kinds of cancer cells? One specific type, called
cancer stem cells (CSC), is catching the eye of drug
developers. Discovered more than a decade ago, CSCs
are hypothesized to be the drivers of cancer growth
and metastasis. Let’s find out the story behind this
potential root cause of cancer and explore the pipeline of
therapies targeting this destructive cell with many faces.

CSC PRIMER
Regular stem cells are prized for their ability to make
copies of themselves (self-renew) without actually
becoming a specific cell type. Rather, they maintain the
potential to become a specialized cell type—say, a heart
or liver cell—in response to a certain mix of chemical The image above shows the difference between using CSC specific
signals. CSCs share the same characteristics: therapy versus conventional cancer therapy. By eliminating the
yellow cancer stem cell from a tumor, the cancer is cut off from
• Cancer stem cells can self-renew: CSCs can continue its source of proliferation, leading to tumor regression.
to divide and produce identical copies of themselves.
• Cancer stem cells can differentiate: CSCs can become MONOCLONAL ANTIBODIES
specialized and develop into multiple cell types that
Stemcentrx (South San Francisco, CA) is developing
make up tumors.
Rova-T, an antibody-drug conjugate that targets both
Cancer stem cells have been identified in most tumor cells and cancer stem cells. Antibody-drug
human tumors. conjugates use the ability of an antibody to recognize
Where do CSCs come from? Most tissue types within a single protein target to deliver a toxic drug directly
our bodies have a collection of regular stem cells that to cells which have the target on their surface. Rova-T
remain in a self-renewing, non-specialized state—they targets the protein DLL3, found on the surface of both
are not carrying out the function of any specific tissue. CSCs and tumor cells. When tested in mice that received
Tissue damage activates those stem cells to develop tumor grafts from small cell lung cancer patients, Rova-T
into replacement cells for that specific tissue. Now, if eliminated both tumor cells and CSCs, suggesting the
a mutation occurs in one or more genes involved with drug may be successful in treating the initial cancer and
cell division, those regular stem cells could morph into preventing recurrence in humans. Rova-T is currently in
cancer stem cells. The opposite may be true too, because Phase II clinical testing.
some fully developed cancer cells acquire mutations OncoMed Pharmaceuticals (Redwood City, CA) is
which could cause them to revert back to a stem-like targeting CSCs with their lead candidate, a monoclonal
state. Since all kinds of stem cells have a relatively long antibody named demcizumab, currently in Phase II
lifespan, the odds of mutation accumulation increases, clinical testing for pancreatic cancer and non-small cell
which supports the idea of CSCs driving cancer. lung cancer. Demcizumab aims for tumor cells and CSCs
CSCs are different enough from actual tumor cells with the surface protein DLL4, which activates white
that many treatments which target and kill tumor cells blood cells to destroy those cancer cells. DLL3 and DLL4
don’t adequately destroy cancer stem cells. Even if the are part of the same family of proteins, and both play

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 Kinase (FAK) pathway. (FAK) pathway. When FAK is
overactive, cells lose their ability to respond to signals
that activate cell death. Verastem (Cambridge, MA) has a
small molecule FAK inhibitor, VS6063, in Phase II clinical
studies for non-small cell lung cancer.

FUSION PROTEINS
Stemline Therapeutics (New York, NY) created a fusion
protein that targets the IL3 receptor, which is present on
both tumor cells and CSCs. A fusion protein is a single
protein that combines characteristics of two different
proteins to fight disease.
Stemline’s product, SL-401, combines the IL3 protein
a role in activating a signaling pathway inside of cancer
with the diphtheria toxin protein. IL3 normally functions
cells that promotes their development.
to activate white blood cells as part of the immune
response. In SL-401, this function is not relevant; it is
SMALL MOLECULE INHIBITORS
simply being used to attach the diphtheria toxin to
Boston Biomedical (Cambridge, MA) is tackling the CSC cancer cells and CSCs that have the IL3 receptor on their
signaling pathways that drive CSC development with the surface. These cancer cells then “take up” or internalize
small molecule drug napabucasin. Currently in Phase III SL-401. Once inside, the diphtheria toxin is released,
clinical studies for gastric/esophageal adenocarcinoma, killing the cell. SL-401 is in Phase II clinical testing for
colorectal, and non-small cell lung cancer, napabucasin blastic plasmacytoid dendritic cell neoplasm, and in
inhibits a protein called STAT3. STAT3 turns on genes Phase I/II for acute myeloid leukemia.
that promote cell growth and development. Normally,
Targeting cancer at its stem cell source offers new hope
STAT3 activation is very tightly regulated, occurring only
for longer term, progression-free cancer survival. Time
in response to specific cues from the cell’s environment.
will tell, but hopefully one of the many CSC-targeting
In cancer stem cells, STAT3 is active all the time, so
therapeutics — monoclonal antibodies, small molecule
inhibiting STAT3 may prevent the transformation of CSCs
inhibitors, and fusion proteins — in development will one
into tumor cells.
day soon be clinical reality.
Another cell-signaling pathway thought to be involved in
CSC development and activation is the Focal Adhesion

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POST-TRANSLATIONAL
MODIFICATIONS IN BIOTECH Life Science Training from Industry Experts

BASIC SCIENCE MEETS APPLICATION PHOSPHORYLATION &

“Post-translational modification” may not be a term you DISEASE: CANCER
hear every day, but it is critically important to the biotech Phosphorylation is important in regulating cell growth
industry—from understanding how cancer develops to and division.
producing biologics. Cell growth and division are regulated by growth
Breaking the term down, we know that: factor receptors, which are proteins found on all
• “post” means “after.” cells. Growth factor receptors pass through the cell
membrane and have an external and internal domain.
• “translation” is “the cellular process of This transmembrane protein allows the cell to take in
making proteins.” messages from the outside of the cell and pass them on
• “modification” means “change.” to the inside of the cell.
So, a post-translational modification (PTM) simply means Once the external domain binds a growth factor signal,
that after a cell makes a protein, a change is made to an internal signaling cascade begins. The cascade usually
that protein. involves the addition of phosphate groups by kinases to
a succession of proteins. The cascade ends when the last
As you know, DNA provides the instructions to the
activated protein enters the cell nucleus and instructs
cell to make a protein, however, that is not the whole
the DNA to make the proteins required for cell division.
story. Often modifications are necessary for the protein
to fold into its correct shape and function properly. Since cancer is uncontrolled cell growth, many different
These changes occur during the process of post- types of cancer are associated with problems in signaling
translational modification. cascades. For example, a mutation in one of the kinases
may cause it to be permanently “on.” This non-stop
In this WEEKLY, we’ll explain the most industry-relevant
activation means the DNA is always getting the signal
post-translational modifications and examine how PTMs
to make proteins that cause the cell to grow and divide.
impacts everything from biologics to biomanufacturing.
This constant cell division eventually results in a tumor.
THE BIG TWO: PHOSPHORYLATION There are 28 FDA-approved kinase inhibitors on the
& GLYCOSYLATION market. A few of the recent approvals include:
Two of the most important PTMs are phosphorylation • Alecensa (Roche, Basel, Switzerland), approved in
and glycosylation. 2015 for non-small cell lung cancer.
Phosphorylation is the addition of a negatively charged • Tagrisso (AstraZeneca, Cambridge, England),
phosphate group to a protein. This phosphate changes approved 2015 for non-small cell lung cancer.
the shape of the protein; in most cases, this shape
change turns the protein “on” from an “off” state.
The enzyme that adds the phosphate group is called
a kinase and kinases play a critical role in different
signaling pathways.
Glycosylation is the addition of a sugar group
(carbohydrate) to a protein. The sugar ensures proper
protein folding and increases protein stability. The
enzyme that adds the sugar group is called glycosylase.
As we’ll discuss below, glycosylation is critical for
monoclonal antibody function. In fact, all antibody
therapeutics must be produced in mammalian cells
and not bacterial cells, because mammalian cells have After the binding of the growth factor to the receptor, the
glycosylase and bacteria do not. growth factor signal activates the the first kinase.

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 glycosylation enzymes, changing the structure of the
sugars attached to the mAb.

PTM IN BIOMANUFACTURING: SMARTAG

Speaking of cell lines, Catalent (Emeryville, CA) has
engineered another type of cell line to produce a
desirable post-translational modification. Unlike
glycosylation, this modification is not common in nature;
rather it is a clever way to create monoclonal antibodies
that can be used for the targeted drug delivery of small
molecules, toxic chemotherapeutics, peptides, RNA,
DNA, or other proteins.
Dubbed SMARTag, the cells are engineered to
overproduce an enzyme called “formylglycine-generating
The signaling cascade involves kinases adding phosphate groups
to the succession. Once the domino effect meets the nucleus, the
enzyme” (FGE). FGE converts the amino acid cysteine to
instructions to initiate cell growth and division are received. the amino acid formyglycine, which is not normally found
in proteins. Formylglycine contains an aldehyde group,
• Cotellic (Exelixis and Genentech, South which can be used to attach whatever molecular entity
San Francisco, CA), approved 2015 for needs to be delivered to a diseased cell. The specificity
advanced melanoma. of this reaction ensures that attachment occurs only
at the formylglycine sites, creating a uniform, stable
GLYCOSYLATION & DRUG product. SMARTag technology is a type of antibody-drug
DEVELOPMENT: MABS conjugate technology (ADC).
Glycosylation is of particular importance to the From glycosylation to phosphorylation, post-
shape, stability, and function of monoclonal antibody translational modifications have a significant impact
therapeutics (mAbs). Many mAbs approved for cancer on protein function, illustrating once again the complex
work by recognizing and binding to a protein on the nature of proteins and protein-based therapeutics.
surface of a tumor cell. This binding attracts white By fully understanding PTM, drug companies can
blood cells, such as macrophages, to attach to the mAb use this process to their advantage to create more
and destroy the antibody-tumor cell complex. Because effective products.
the white blood cells recognize mAbs based on their
glycosylation pattern (i.e. the specific type of sugar
groups attached and the location of the attachments),
changes to the glycosylation pattern may increase or
decrease the drug’s efficacy.
One prime example is Roche’s (Basel, Switzerland)
leukemia drug Gazyva. Gazyva has significantly better
clinical efficacy than Roche’s earlier drug Rituxan,
even though both drugs target the same protein on
blood cancer. This is because Gazyva has a different
glycosylation pattern than Rituxan, making the antibody-
tumor complex more recognizable by macrophages.
This new glycosylation pattern was achieved by
producing Gazyva in a mammalian cell line that has been
genetically engineered to overproduce two alternative
The Y-shaped SMARTag antibody above shows how the
formylglycine containing aldehyde group can be used to
attach a drug after administering a chemotherapeutic agent,
turning it into a Y-shaped antibody-drug conjugate.

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THE FIRST THREE-PARENT BABY Life Science Training from Industry Experts

BREAKING DOWN THE baby. The resulting child has DNA from two different
SCIENCE & SCRUTINY women and one man.

“World’s First Three-Parent Baby” made headlines in LEIGH SYNDROME IS THE REASON
publications ranging from Nature to CNN last week. This
The three-parent boy’s parents sought out the TPIVF
human interest story is of a baby boy born in Mexico
technique because they had already lost two children
with genetic material from three different parents,
to a rare, mitochondrial DNA-linked disease called Leigh
achieved by a technique known as three-parent in vitro
syndrome. Referred to as a “neurometabolic disorder,”
fertilization (TPIVF). In this issue, we will explain why
Leigh syndrome affects the central nervous system.
TPIVF is used and examine the science—and scrutiny—
Symptoms usually begin within a year of birth and
surrounding this controversial baby-making method.
include failure to thrive, low muscle strength, and lack of
REPLACING MITOCHONDRIAL DNA IN IVF control over movements. Death by cardiac or respiratory
The rationale behind TPIVF lies with mitochondria— failure usually occurs within a few years.
what many of us remember from high school biology
as the “powerhouse” of a cell. Recall mitochondria are
CYTOPLASMIC TRANSFER
the organelles that convert glucose into the energy
STARTED IT ALL
our cells use to do work. What you may not remember TPIVF has some precedent in a procedure known as
is mitochondria have their own DNA that is inherited cytoplasmic transfer. In the late 1990s, this experimental
maternally. When an egg is fertilized, the mitochondria procedure was used to help women, whose fertility had
from the egg get passed on in subsequent rounds of cell declined, to conceive.
division, becoming a part of every cell in the developing The prospective mother’s egg was injected with a
baby. A woman with defective mitochondrial DNA passes small amount of cytoplasm from another woman’s egg
this trait onto her child. However, with the advent of before fertilization. Cytoplasm is the liquid portion
TPIVF, these mutations can be weeded out of a family’s of a cell outside of the nucleus that also happens to
genetic landscape. contain mitochondria. The exact mechanism by which
During TPIVF the nuclear DNA is removed from the egg cytoplasmic transfer enables pregnancy is not clearly
of the afflicted mother. This nuclear DNA is transferred defined, but many doctors suspect some cases of
into a donor egg whose own nuclear DNA has been infertility are caused by damaged mitochondria.
removed, but whose mitochondrial DNA remains Twenty-four women achieved pregnancy via cytoplasmic
intact. The resulting donor egg is implanted into the transfer two decades ago. However, the FDA had safety
prospective mother, and with fingers crossed, that donor concerns, and the practice was abandoned when fertility
egg (once fertilized with a sperm) develops into a healthy clinics were required to file an IND to continue the
procedure. Cytoplasmic transfer is still not approved
by the FDA, and this policy is credited with creating a
market for reproductive tourism abroad.

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EXAMINING GERMLINE MODIFICATION with the restriction that it be limited (at least initially)
to male offspring because men cannot pass down
TPIVF has the potential to become a routine medical
mitochondrial DNA. The panel also recommended
procedure, but not without some scrutiny involving
following any children born of this technique for several
germline modification. In more simple terms, germline
years and sharing data with the public. The procedure
modification is the genetic changes made to sperm or
was approved in the UK in 2015 without restriction.
egg cells. Any genetic changes made to these sex cells
will be passed down to all resulting generations, which COCKTAIL FODDER: ONLY 37 GENES
may have unforeseen risks and consequences.
Although it is correct to say that a baby conceived by
The world’s first “three-parent baby” was born in Mexico TPIVF has genetic material from three different parents,
to Jordanian parents with assistance from a New York the vast majority (~99.9%) of that baby’s genetic material
City-based fertility doctor. Why not the United States? will come from the nuclear DNA of the original egg and
Because the FDA has yet to officially approve the sperm. Mitochondrial DNA codes for only 37 genes,
technique. In February 2016, a U.S. National Academies whereas nuclear DNA codes for ~21,000 genes.
of Science panel recommended approving the method

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A BONE TO PICK WITH OSTEOPOROSIS Life Science Training from Industry Experts

THE RACE TO BE THE NEXT currently on the market, and which ones in the pipeline
BIG BONE DRUG are threatening to become the new gold standard for
osteoporosis care.
Amgen’s recent positive Phase III results for an antibody
drug is putting it neck and neck with Radius Health’s BONE FORMATION 101
peptide — the latest in a batch of osteoporosis therapies
Bone formation and maintenance is a complex process
edging their way to the market. The current widely
that relies on many different interacting factors but
prescribed generic — bisphosphonate — only works to
is driven by two specific cell types: osteoblasts, which
slow the loss of bone, while these newer drugs add to
lay down bone tissue, and osteoclasts, which degrade
the therapies that aim to rebuild.
bone tissue. This process may be activated when bone
Osteoporosis is the loss of bone density which causes remodeling is required, such as in response to a fracture.
bones to become weak, brittle, and easily broken. In Osteoclasts break down the damaged bone tissue by
healthy people, bone is constantly being broken down secreting an acidic substance in a process known as
and replaced. When bone is broken down more quickly bone resorption, and then osteoblasts produce new
than it is replaced, osteoporosis occurs. As the disease tissue for bone formation. During childhood, formation
progresses, patients become more vulnerable to broken exceeds resorption; as people age, resorption begins to
bones, especially in the hip, spine, and wrist. In advanced exceed formation.
cases, even minor falls or bumps can result in a fracture,
Other factors that influence bone formation and
leading to loss of mobility.
maintenance include calcium, vitamin D, the hormone
According to the National Osteoporosis Foundation, estrogen, and weight-bearing exercise. Calcium is
osteoporosis-related fractures number as many as the mineral that provides bone with its hardness and
two million per year in the US, with related costs at $19 strength. Because calcium plays a role in other critical
billion. These numbers are expected to climb to three metabolic processes like muscle contraction, diets
million and $25.3 billion, respectively, by 2025. Let’s inadequate in calcium may trigger the activation of
break down how bone loss occurs, which therapies are bone resorption in order to release supplies of this
vital mineral into the bloodstream. Making sure that we
absorb the mineral from the food we eat is necessary to
keep up with demand, and vitamin D is essential to the
calcium absorption process.
Weight-bearing exercise such as walking, jogging, or
weight-lifting help to build and maintain bone mass.
When stress is put on bones from weight-bearing
exercise, they respond by activating osteoblasts to
better withstand the stress which ultimately increases
bone density.
It’s long been known that estrogen protects women
from bone loss. In the past decade, researchers at
the University of Buffalo pinpointed the reason
why: estrogen protects osteoblasts by inhibiting an
enzyme that would normally trigger apoptosis—
programmed cell death—in the osteoblast cells. This is
why post-menopausal woman have the highest rates
of osteoporosis.

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ACTIVATING APOPTOSIS at preventing vertebral fractures as Forteo, and more
effective at preventing other types of fractures. Radius
The most common treatment for osteoporosis today is
submitted a new drug application to the FDA in May
bisphosphonate with the most widely prescribed drug
2016. Like Forteo, abaloparatide is an anabolic, or bone-
being Fosamax (Merck; Kenilworth, NJ). Bisphosphonate
growing, treatment.
drugs activate apoptosis (cell suicide) in osteoclasts,
and have been clinically shown to decrease fractures THWARTING RANKL
of the wrist, spine, and hip of at-risk postmenopausal
Another FDA-approved drug for osteoporosis is Amgen’s
women. These bone-preserving drugs may have some
(Thousand Oaks, CA) Prolia. Prolia is a monoclonal
unpleasant gastrointestinal side effects, however, and
antibody that works by inhibiting RANKL, a protein
their long-term use is associated with the development
on the surface of osteoclasts. By impeding RANKL,
of low-impact femoral fractures. Thus the race is still on
osteoclast activation is diminished.
to find new osteoporosis drugs.
Amgen recently announced positive Phase III results for
PTH, THE REMIX a second antibody osteoporosis drug, romosozumab.
Forteo (Eli Lilly, Indianapolis, IN) is another popular Romosozumab works by inhibiting a protein called
treatment for osteoporosis. It is a recombinant version sclerostin, which hampers bone formation by
of the parathyroid hormone (PTH). While chronically high osteoblasts. By suppressing the inhibitor, the drug
levels of parathyroid hormone will activate osteoclasts, appears to make bone-producing osteoblasts more
a once-daily injection of PTH can activate osteoblasts active. A second company, OsteoGeneX (Kansas City,
more than osteoclasts, thereby increasing bone growth. KS), is also developing a small molecule inhibitor of
sclerostin. Sclerostin inhibitors are bone-building agents.
Abaloparatide, by Waltham, MA startup Radius Health,
is a peptide analog of human parathyroid-related As the population ages, osteoporosis will become
protein. It is a naturally occurring peptide hormone an even more prevalent problem. These new bone-
that has a similar effect as parathyroid hormone. In strengthening drugs provide hope for good health and
Phase III clinical trials, abaloparatide was as effective mobility for at-risk older adults.

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DMD MAKES THE CUT Life Science Training from Industry Experts

THE SCIENCE BEHIND SAREPTA’S TERMS OF THE WEEK: INTRON & EXON
HOTLY DEBATED ANTISENSE DRUG DNA and RNA are made up of segments of introns and
Sarepta’s (Cambridge, MA) Duchenne muscular exons. An intron does not code for a protein. An exon
dystrophy drug Exondys 51 crossed the finish line earlier does code for a protein. Said another way, exons make
this week, with a conditional stamp of approval by the up genes and introns separate genes.
FDA. This hotly debated regulatory result offers new When DNA is first “read” by cellular enzymes, the result
hope for patients and families whose previous treatment is a pre-messenger RNA (pre-mRNA). This pre-mRNA
options only managed the descent of the disease. contains both introns and exons. More cellular enzymes
Affecting one in approximately 3,500 newborn baby join the party: some cut out the introns and others splice
boys worldwide, Duchenne muscular dystrophy (DMD) the exons together to form mRNA. It is this mRNA that
is caused by a mutation in the gene that codes for contains instructions the cell needs to make a protein.
dystrophin, resulting in a lack of the protein. DMD is The Exondys 51 namesake is partly derived from
characterized by damaged muscle cells, progressive the word exon. Bet you can guess where “dys 51”
muscular weakness, and serious medical problems, comes from!
including significant impairment of the heart and lungs.
Typical life expectancy for a child with DMD is 25 years.
Today’s WEEKLY is a primer on DMD and the science
driving Exondys 51.

MUSCLE GLUE
DMD patients produce no functional dystrophin
protein—a protein so important that its loss is fatal.
In muscle cells, including cardiac muscle, dystrophin is
part of a complex that works together to strengthen and
protect muscle fibers from injury during contraction
and relaxation cycles. Dystrophin’s job is to connect Introns are the segments in between the different colored exons
shown above. Introns are discarded during splicing, leaving only
the proteins that make up the muscle cell structural the exons to make up the final protein product.[/caption]
framework (the cytoskeleton) with the network of
proteins that surround each cell (the extracellular
matrix). For this reason, dystrophin is sometimes
referred to as the “glue” that holds muscle cells together.

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EXONDYS 51 EXPLAINED EXON SKIPPERS IN DEVELOPMENT
Exondys 51 is a first in class biologic. It is an antisense All cases of DMD are caused by mutations in the
therapy, meaning it is a short, synthetic RNA analogue dystrophin gene, a large gene containing 79 exons.
also referred to as an antisense oligo. When Exondys 51 Mutations amenable to exon 51 account for about 13% of
enters a patient’s cell, it binds to exon 51. This binding all DMD cases. Sarepta estimates it can develop exon-
causes the enzymes that convert pre-mRNA into mRNA skipping technology for 80% of the DMD population and
to skip over that exon so that exon 51 is not included in has therapies targeting exon 45 and exon 53 in Phase I
the mRNA. The resulting mRNA produces a protein that and II clinical testing, respectively.
is a truncated version of the normal dystrophin protein. Exondys 51 isn’t the only exon-skipping drug shaking up
the world of biotech. Ionis Pharmaceuticals (Carlsbad,
CA) has an exon-skipping treatment in Phase III clinical
trials for the treatment of the rare disease spinal
muscular atrophy, and Wave Life Sciences (Cambridge,
MA) has several exon-skipping treatments in preclinical
development, including drugs that target spinal muscular
atrophy and DMD.

COCKTAIL FODDER: A
RECIPE FOR SPLICING
Alternative splicing is the process of including some
The exon splicing shown above demonstrates how a mutation exons and excluding others in the final mRNA.
is able to make it into the final protein.[/caption] Alternative splicing is not a random act. This explains
how a single gene can give rise to multiple types of
Interestingly enough, patients with Becker muscular
proteins and why humans can get by with ~20,000
dystrophy (another rare disease) naturally produce the
genes. In fact, the gender of the good ole’ geneticist’s
same truncated dystrophin protein and experience a
favorite subject, the fruit fly, is partially determined
milder form of muscular disease as compared to DMD.
by alternative splicing. It’s been 40 years since the
So, the production of a partially functioning dystrophin
phenomenon of alternative splicing was discovered, and
protein appears to be enough to restore some
we are just now learning to modulate the process.
functionality to damaged muscle cells.

Exon skipping involves an “antisense oligo” shown above that

targets the mutation, rendering it incompatible. The mutation
is now excluded from the makeup of the final protein.

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THE TIGHT JUNCTION’S FUNCTION Life Science Training from Industry Experts

A PATHWAY LIES IN THE THE GAP IN CELIAC

BRIDGE BETWEEN CELLS In patients with celiac disease, consuming the wheat
Most of us remember from high school biology that protein gluten disrupts the barrier function of tight
cells are the basic building blocks of all living things. junctions within the intestinal epithelium. This disruption
Cells join together to form tissues and organs such as occurs when gliadin, a component of gluten, binds
hearts, lungs, and livers, which work together to create to a receptor protein on the surface of intestinal
a functioning human. An adult human body contains epithelial cells, initiating a signaling pathway resulting
trillions of cells, and these cells must coordinate to form in tight junction disassembly. This, in turn, leads to
a cohesive whole. increased intestinal permeability and the activation of
One of the ways in which cells work together is through inflammatory pathways that cause serious damage to
cell junctions, or “intercellular bridges.” These junctions intestinal cells.
consist of multiprotein complexes that anchor cells to Innovate Biopharmaceuticals (Raleigh, NC) is
each other — and in some cases enable communication developing a peptide (short sequence of amino acids)
between them. In this WEEKLY, we’ll zoom in on one drug that appears to reverse the opening of tight
specific type of cell junction known as a tight junction, junctions in celiac patients. It does this by inhibiting the
and examine its role in health and disease. signaling pathways initiated by gliadin binding. Dubbed
larazotide acetate, the drug has successfully completed
TIGHT JUNCTIONS UNRAVELED Phase II clinical studies and is preparing to enter
Tight junctions are found in epithelial cells, or the cells Phase III.
that line the cavities and surfaces of blood vessels
and organs. In addition to holding cells together, THE GAP IN GASTROINTESTINAL
tight junctions form a barrier between adjacent cells, & PANCREATIC CANCER
preventing the passage of various molecules into tissues One of the key protein components of tight junctions
between cells. Anything entering a tissue must actually is a family of proteins called claudins. Ganymed
enter the cells themselves rather than going through Pharmaceuticals (Mainz, Germany) is developing a
any “spaces” around them. Tight junctions enable monoclonal antibody (mAb) therapeutic (IMAB362) that
tighter control over what gets in and out of the tissue by targets claudin 18.2, a subtype of claudin that is only
creating barriers between cells. expressed in mature stomach cells and is overexpressed
The tight junction itself is composed of proteins from in 80% of gastrointestinal adenocarcinomas and 60% of
two neighboring cells that pass through each cell pancreatic tumors. The tumor growth disrupts the tight
membrane and interact with each other outside of the junction function in these cases. Because the protein is
cells, bridging the two cells: so specific to gastrointestinal cells, it makes a good drug
target—the mAb ideally should not impact other tissues
in the body, reducing the potential for toxicity. IMAB362
showed very promising results in a Phase II clinical trial
for gastroesophageal cancer and is preparing to enter
Phase III trials. The drug has also shown promise in
preclinical studies of pancreatic cancer.
In order to ensure optimal efficacy, IMAB362 should
be given to patients who over express claudin 18. To
meet this need, Ganymed is also developing Claudetect
The tight junction visualized. 18.2, a companion diagnostic for IMAB362. Claudetect
determines expression levels of claudin 18.2 to find out

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which patients are most likely to respond to treatment. harmful substances from entering the brain. In patients
It is currently being used to identify patients for with multiple sclerosis, inflammation disrupts the BBB,
Ganymed’s IMAB362 clinical trials. allowing white blood cells to enter the brain and attack
neurons. Brain tumors also disrupt the BBB, resulting
CRITICAL BARRIER: BLOOD- in potentially fatal brain swelling due to excess fluid
BRAIN BARRIER entering the brain.
A great example of a tight junction barrier function is the We often think in terms of what our tissues need, but a
blood-brain barrier (BBB). This barrier is formed by tight closer look at tight junctions reminds us that sometimes
junctions between cells lining the capillaries surrounding what cells and tissues keep out can be just as important
the brain. Although this barrier can present a challenge as what is allowed inside.
to scientists trying to deliver medicines to the brain,
the BBB plays the critical function of preventing most

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ZIKA ON THE BRAIN Life Science Training from Industry Experts

THE LATEST ON THE RISE OF ZIKA GETTING A DIAGNOSIS

The spread of the Zika virus is raising alarm bells in Developing a new diagnostic test and gaining FDA
the United States, and Congress has yet to pass any approval can take years—even a decade or more. In the
legislation to combat the fight. Zika infections reported case of a potential public health emergency, the FDA
in Miami confirm the virus’ stateside arrival, pushing may grant Emergency Use Authorizations to make
residents to take measures to protect their pregnancies diagnostic tests available more quickly. In the past six
from potentially devastating effects. In our previous months, the FDA has used this maneuver to authorize
issue, we explored the underpinnings of Zika: it is linked tests to detect either RNA (Zika’s genetic material) in
to microcephaly, in which babies are born with brains early infections or to find the presence of anti-Zika
significantly smaller than normal. antibodies in blood. The RNA-based tests may detect
New reports state the disease is sexually transmissible, the virus much earlier during infection, but may have a
persisting in those infected for up to six months. This has higher incidence of a false negative, so antibody tests are
quickly turned into a public health crisis, with the FDA advised to confirm the diagnosis. The RNA-based tests
recently recommending universal testing of all donated granted emergency authorization include:
blood across the US. As the wrath of Zika continues to • Roche Molecular Systems’ (Rotkreuz, Switzerland)
migrate onward, today’s WEEKLY showcases the latest LightMix Zika rRT-PCR test.
developments surrounding this mosquito-borne virus. • Luminex Corporation’s (Austin, TX) xMAP MultiFLE
Zika RNA assay.
ZIKA ON THE BRAIN
• Siemen’s Healthcare Diagnostics’ (Erlangen,
Two recent studies on the basic science of Zika
Germany) Versant Zika RNA 1.0 Assay.
discuss how the virus may impact brain cells called
human neuronal stem cells—the same cells found in • Viracor-IBT Laboratories’ (Lee’s Summit, MO) Zika
developing fetal brains. Yale University (New Haven, Virus RT-PCR Test.
CT) researchers demonstrated that the Zika virus infects • Hologic’s (Bedford, MA) Aptima’s Zika Virus Assay.
lab-grown human neuronal stem cells, disrupting their • Altona Diagnostics’ (Hamburg, Germany ) RealStar
reproduction and leading to cell death. By interacting ZIka Virus RT-PCR Kit.
with a cellular protein called TBK1, Zika prevents cells
• Focus Diagnostics’ (Cypress, CA ) Zika Virus RNA
from organizing during cell division—suggesting a cause
Qualitative Real-Time RT-PCR.
and effect for newborn microcephaly.
• Center For Disease Control’s (Atlanta, GA) Trioplex
Adult brains maintain reserves of what are referred to
Real-time RT-PCR Assay.
as “neural progenitor” cells. These are stem cells that
have begun the process of differentiating into neurons, The antibody-based tests granted emergency
and are found in the adult brain in specialized locations authorization include:
called “niches.” These progenitor cells play an important • InBios International’s (Seattle, WA) ZIKV Detect IgM
role in learning and memory. Researchers at Rockefeller Capture ELISA.
University (New York City, NY) and La Jolla Institute for
• Center For Disease Control’s Real-Time RT-
Allergy and Immunology (La Jolla, CA) demonstrated
PCR Assay.
the virus infected adult neural progenitor cells in a
mouse model in much the same way as seen with fetal
THERAPIES ON THE RISE
stem cells. Since the brain is already fully developed, the
impact may be much more subtle than the microcephaly While the current recommended treatment is rest and
cases seen in infants. Zika could potentially have an hydration, researchers at Yale found the hepatitis C virus
unappreciated impact on adult learning and memory. (HCV) drug Sovaldi (Gilead; Foster City, CA) appeared

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to prevent Zika infection of cultured stem cells. Sovaldi
or its derivatives may be a potential therapeutic or
preventative treatment after additional clinical testing.
This may seem surprising, but despite causing very
different diseases, it turns out HCV and Zika come
from the same family of viruses. They share structural
similarities: both have a single strand of RNA as their
genetic material, and both rely on an enzyme called an
RNA-dependent RNA polymerase (RdRp) to replicate.
Although the two RdRps are not identical, it is possible
that since Sovaldi inhibits this enzyme in HCV it may also
inhibit the RdRp in Zika.
Researchers at SRI International (Menlo Park, CA) are
screening existing anti-viral compounds in hopes of development of a broad-spectrum antiviral drug for Zika
unearthing a potential Zika therapeutic. In this case, the virus,” says Beylkin. “We are going to need a combination
chemical library contains more than 3,000 members, of new antiviral drugs and vaccines to successfully
including hundreds of anti-influenza compounds control this emerging threat.”
specifically targeting the influenza RdRp. Although not
in the same family as Zika, influenza is also an RNA A PREVENTION CONVENTION
virus and relies on a RdRp for replication. According There are still no vaccines on the market, but Inovio
to Dr. Diane Beylkin, a lead scientist for the project, Pharmaceuticals (San Diego, CA) recently announced
the RdRp enzyme makes an especially good target a second round of clinical testing in Puerto Rico for its
because it mutates less than other viral proteins, making DNA-based Zika vaccine candidate. Its first round in June
resistance less likely to emerge. Dr. Beylkin’s team is was the first test of a Zika vaccine on humans. Earlier
developing a novel screening assay using a modified in August, the National Institute of Health (Bethesda,
Zika virus which can replicate but is unable to assemble MD) also began clinical testing of a DNA-based Zika
into new virus particles, enabling the measurement of vaccine. Several other companies are conducting
replication (RdRp) inhibition without actually producing preclinical studies of Zika vaccines, including Takeda
new virions. “It is very exciting to be collaborating with (Osaka, Japan), GlaxoSmithKline (Brentford, UK), and
SRI’s experts on vector-borne diseases, biochemical Sanofi (Paris, France).
assay development, and antiviral drug discovery on the

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THE LONG AND SHORT OF AMD Life Science Training from Industry Experts

A SPOTLIGHT ON ONE OF THE AMD EXPLAINED

MOST PREVALENT EYE DISEASES AMD is caused by damage to the macula, resulting in
It’s easy to take our ability to see for granted. On a day- blurred vision progressing to vision loss in the center of
to-day basis, we rely on vision to process information the patient’s field of vision. The exact cause of macular
and to navigate the world without giving it a second damage is not known, but AMD is associated with the
thought. For the millions of people affected by eye buildup of proteins and lipids just beneath the retina.
disease, vision loss, and blindness, impaired vision These deposits are referred to as “drusen,” and are a
is a daily reality that changes how they interact with normal part of aging but are increased in AMD. As the
the world. disease progresses, vision loss increases. Advanced AMD
Age-related macular degeneration (AMD) is one of the is classified into two types: neovascular (wet) or atrophic
most common eye diseases, affecting 13.4 million people (dry).
in the United States and is the leading cause of vision
loss in those over the age of 50. In this WEEKLY, we get
THE WET
out our funduscope to peer into the science behind this In wet AMD, macular damage is caused by an infiltration
devastating disease of the retina. of blood vessels into the retina. These abnormal vessels
often leak fluid and blood, causing damage. Wet AMD
TERM OF THE WEEK: RETINA progresses quickly, and can lead to rapid loss of central
The retina is composed of light-sensitive nervous tissue vision if not treated. About 10% of advanced AMD cases
which forms the thin membranous lining in the rear two- are wet.
thirds of the eyeball. Its job is to convert images from the FDA-approved treatments available include Lucentis
optical system of the eye into electrical impulses that are (Genentech; South San Francisco) and Eylea (Regeneron
sent along the optic nerve to the brain. Pharmaceuticals, Tarrytown, NY). Both work by
“mopping up” excess vascular endothelial growth factor
(VEGF), which causes the excess abnormal blood vessel
growth. Lucentis is a monoclonal antibody specific for
VEGF. Eyelea is a fusion protein consisting of the VEGF-
receptor fused to the constant region of an antibody for
stability. Similar to a mAb, this construct is highly specific
for VEGF and binds to the abnormal blood vessels before
it binds to any other blood vessel receptors found in the
eye. These VEGF-blocking treatments have proven to be
very effective at stopping the progression of AMD, but
they do not cure the disease.
Ophthotech (New York, NY) is developing a new
treatment, Fovista, for wet AMD that targets platelet-
The anatomy of the eye (The National Eye Institute).
derived growth factor (PDGF), a second growth factor
critical for the formation and maintenance of new blood
Considered an extension of the brain, the retina is vessels. Currently in Phase III clinical studies, Fovista in
formed embryonically from brain tissue and connects combination with a VEGF inhibitor appears to be more
directly to the brain via the optic nerve. The macula is effective than the VEGF inhibitor alone. Fovista is a DNA-
the small central area of the retina and is the area of aptamer drug, meaning that it is a short DNA sequence
acute central vision. selected for its ability to bind and inhibit PDGF.

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 progression. There are a number of drugs in clinical
development targeting the complement pathway:
• Roche’s (Basel, Switzerland) Phase III clinical studies
on lampalizumab. Lampalizumab is a Fab, or antigen-
binding fragment of a monoclonal antibody, and
targets complement factor D.
• Apellis Pharmaceuticals’s (Crestwood, KY) peptide
APL-2 in Phase II development.
• Novartis’s (Basel, Switzerland) mAb Tesidolumab/
LFG316 in Phase II.
• Ophthotech’s (Princeton, NJ) DNA aptamer Zimura
in Phase IIa.

THE GENETICALLY BASED

Most cases of macular degeneration are associated
with aging. However, juvenile forms, such as Stargadt
disease, are associated with several different genes and
symptoms of blurred central vision which start to occur
THE DRY in adolescence, usually progressing to legal blindness.
Also referred to as macular dystrophy, all varieties of
Dry AMD involves a gradual breakdown in the light-
juvenile macular degeneration are an inherited disease
sensitive cells of the macula. The dry variety progresses
with no current cure.
much more slowly than wet and accounts for about
90% of AMD cases. Currently, there are no treatments As the rate of the aging population grows, projections
available for dry AMD, although some studies have point to 20 million suffering from AMD by 2020. The
suggested that taking high doses of antioxidants need to discover therapies for wet and dry AMD is in
including C and E vitamins, copper, zinc, and beta- high demand. As the pipeline evolves, we look to drug
carotene may slow progression. development to treat our treasured sense of sight.
Inflammation and the activation of the complement
pathway is associated with drusen buildup and dry AMD

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VACCINES: SCHOOLING THE HERD Life Science Training from Industry Experts

VACCINATION NATION THE ABCS OF VACCINES

Back to school means shopping for new school supplies, There are a few ways to create a vaccine and below we
adjusting to a new schedule and making sure all required list the most common methodologies.
vaccinations are up to date. • Inactivated vaccines: The most obvious type is the
Every state requires school-age children to be vaccinated inactivated vaccines—the use of heat or chemicals
against certain infectious diseases including tetanus, to kill the pathogen. Inactivated vaccines produce a
hepatitis B, measles, mumps, rubella, polio, pertussis dampened immune response in comparison to other
(whooping cough), and chicken pox. Vaccination policies vaccine methods and often require “booster” shots.
are highly effective at eliminating many types of sickness Inactivated vaccines include polio, influenza, and
from the most perfect incubator—the classroom. pertussis vaccines.
As with anything “required,” there are loopholes. • Live, attenuated vaccines: A second category
Certainly, medical exemptions are necessary for the is live, attenuated vaccines, which simply means
immunocompromised or children allergic to vaccine weakened (not killed) pathogens that are unable
components. However, there is a vocal minority of to cause disease. Attenuation occurs by a process
parents who wish to skip out on vaccinations due called “passage,” or growing viruses at temperatures
to religious or personal convictions. This presents a slightly lower than the human body or in cells
problem, as witnessed in California during the 2014 different from the human host cells. Under these
measles outbreak and the 2010 whooping cough conditions, the virus accumulates mutations that
outbreak. These two avoidable incidents pushed make it better able to survive in a new environment,
California to pass one of the most stringent vaccination but when injected into a human, it is no longer
mandates in the country this past summer—Senate virulent. Attenuation may also be more direct and
Bill 277. occurs when genes associated with pathogenesis
In this WEEKLY, we’ll go to the chalkboard to learn the or replication are removed in the lab. In general,
basic science of the vaccines. attenuated vaccines induce a strong and long-lasting
immune response. Examples of live, attenuated
THE BASIC SCIENCE vaccines include measles, mumps, rubella, chicken
pox, and polio.
The idea behind vaccination is simple: by exposing
someone’s immune system to a harmless version of a • Subunit vaccines: In some cases, just one protein
pathogen, we can train it to recognize and respond to from the virus can be enough to induce an immune
the bug in the wild. Our immune system creates memory response. These are called subunit vaccines and are
cells after an initial exposure to a virus: memory cells are typically made using recombinant DNA techniques to
then ever-ready to spring into action and attack disease. produce the desired protein. Advantages of subunit
vaccines include easier production and a better
Vaccination is required because creating these pathogen-
safety profile for patients. Examples of subunit
specific memory cells takes a few weeks—a length of
vaccines include pertussis and hepatitis B.
time that is long enough for a virus to do serious damage
to the body. • DNA vaccines: DNA vaccines are the next frontier in
vaccine development. Rather than delivering a whole
pathogen or pathogen subunit, DNA vaccines deliver
just a gene. Once inside the body, the patient’s own
cells reproduce the pathogenic protein. If successful,
this technology would mimic a natural infection
and elicit a strong immune response. The technical

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 challenge that remains to be solved is the delivery of The structure of those two proteins changes every
the pathogenic gene. Ichor Medical Systems (San season due to a high mutation rate. Once the structure
Diego, CA) and Inovio Pharmaceuticals (San Diego, changes, the immune system no longer recognizes it,
CA) are both developing electroporation-mediated and the body must be retrained. This is also why the
DNA delivery systems to solve this problem. vaccine is only 60-70% effective—when formulating each
year’s vaccine, scientists attempt to predict the influenza
TERM OF THE WEEK: HERD IMMUNITY strains that will be circulating in winter, and they are
Herd immunity means a significant portion of a seldom 100% correct.
population has immunity to a particular pathogen. There For many years, scientists have talked of producing
is little opportunity for an outbreak, so even those who a universal flu vaccine. Recent advances in vaccine
cannot be vaccinated such as immunocompromised technology have led to some promising developments.
individuals, pregnant women, or newborn babies One approach involves formulating the internal
are unlikely to become infected despite their proteins with compounds known as “adjuvants,” or
unprotected state. immune response stimulators. Primed by the adjuvant,
the immune system produces antibodies against the
associated protein—an approach used by VaxInnate
(Cranbury, New Jersey). Clinical testing demonstrates
that an adjuvant-based vaccine targeting one of the
virus’ internal, more stable proteins, is capable of
boosting patient’s response to current vaccines. It has
yet to be tested as a stand-alone vaccine.
BiondVax (Ness Ziona, Israel) has identified nine
epitopes—short sequences of proteins that elicit an
immune response—that do not vary much between
different strains of the virus. These sequences were
combined to make one recombinant protein referred to
as Multimeric-001 (M-001). The hope is this combination
of epitopes will invoke a strong immune response which
will be protective over several flu seasons. M-001 is in
Phase II clinical testing.
Researchers at Crucell Vaccine Institute of Janssen
Pharmaceuticals (Leiden, Netherlands) have discovered
an antibody that recognizes and binds a portion of the
HA protein that doesn’t mutate very rapidly. Studies
in animals suggest treatment with this antibody
significantly reduces the amount of active virus present.
Human clinical trials are in the works.

A
A clinically tested and FDA-approved universal flu
breakdown of herd immunity, courtesy of the National vaccine is still years away, but these early results are
Institute of Allergy and Infectious Diseases. promising. The new vaccine would most likely need
to be administered every five or 10 years, rather than
A UNIVERSAL FLU VACCINE? annually—but the real advantage will be its ability to
protect against a range of different influenza strains,
Unlike most other vaccines, you must get the flu shot
inching closer to 100% efficacy. And since most of us
every year in order to be afforded protection. Current
have suffered through a bad case of the flu, we can
flu vaccines work by mounting an antibody response
all agree that a universal flu vaccine cannot come
against two large proteins on the surface of the virus—
soon enough!
hemagglutinin (H) and neuraminidase (N). The catch?

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THE NEXT GENERATION FIGHT AGAINST CF Life Science Training from Industry Experts

TARGETING THE ROOT OF EASILY CONFUSED: POTENTIATORS

CYSTIC FIBROSIS VS. CORRECTORS
Innovative therapies targeting the root cause of cystic CFTR mutations can be divided into two classes: those
fibrosis (CF) hit the market several years ago, but those that lessen the quantity of CFTR proteins reaching the
treatments were only for a subset of CF patients. Now, cell surface, and those that reduce the functioning of
companies like AbbVie and Vertex may have the the proteins reaching the cell surface. Drugs that work
potential to treat a large majority of the population—up by assisting CFTR to fold correctly and reach the cell-
to 90%—according to clinical trial data. In this issue, surface are referred to as CFTR correctors; drugs that
we’ll explain the science of cystic fibrosis and review the enable CFTR to function correctly once it has reached
pipeline of hope for this orphan disease. the cell surface are called CFTR potentiators. These two
different mechanisms of action are used in combination
CYSTIC FIBROSIS PRIMER to fight CF at the cellular level.
Cystic fibrosis is a genetic disease caused by one of
several possible mutations in the gene that encodes the ON THE MARKET: KALYDECO & ORKAMBI
cystic fibrosis transmembrane conductance regulator In 2012, Vertex Pharmaceuticals (Boston, MA) made
(CFTR) protein. The CFTR protein is critical for the headlines by winning FDA approval for Kalydeco,
production of sweat, digestive fluids, and mucus. ushering in the first CF drug to treat the underlying
CFTR is classified as a channel protein—a category of cause of the disease. Kalydeco works by binding to
proteins that create a channel, or tunnel, across the cell the misfolded CFTR protein and increasing its ability
membrane. This specialized gateway allows things to to remain open and functional on cellular surfaces—a
pass through the cell that would otherwise be denied CFTR potentiator. Although highly effective, it is only a
entry or exit. Negatively charged chloride ions use CFTR lifesaver for approximately 10% of CF patients. Kalydeco,
to exit cells, and if CFTR is not functioning correctly, the when used alone, is not helpful to patients whose CFTR
chloride ions build up inside of cells, causing the cells is unable to reach the cell surface due to their type
to retain water. This buildup affects the fluid balance of mutation.
of tissue, resulting in the characteristically thick mucus When Vertex combined Kalydeco with newly developed
seen in the lungs of CF patients, making them vulnerable lumacaftor, a dynamic duo was born. Coined Orkambi,
to fatal lung infections. the combination has the potential to benefit as many as
CF is an autosomal recessive disorder, meaning if an 50% of CF patients. The lumacaftor portion targets the
individual has one functioning copy of the CFTR gene, most common mutation (a single amino acid deletion
they will not develop the disease and are termed in the CFTR protein) responsible for two-thirds of CF
“carriers.” Two copies of the malfunctioning CFTR gene cases. In patients carrying this mutation, the protein
(one received from each parent) means a CF diagnosis. is so misfolded it never makes it to the cell surface.
And while CF is always caused by a mutation in CFTR, Lumacaftor is a CFTR corrector; it works by binding to
many possible mutation combinations have been and stabilizing at least some of the misfolded proteins,
associated with the disease. improving their ability to travel to the cell surface. Once
there, Kalydeco kicks in and improves the function of
Developing new drugs for CF is an example of the
CFTR. Orkambi was approved in 2015 and is effective in
much talked about precision medicine initiative:
approximately 50% of CF patients.
determining which patient populations will best
respond to a particular type of therapy based on their
specific mutations.

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IN THE CLINIC: NEXT GENERATION
CFTR CORRECTORS
The next generation of CFTR correctors offer more hope
to the CF community, with a giant step forward in drug
development. Preclinical testing of next generation CFTR
correctors predict efficacy greater than lumacaftor—and
these are already in clinical trials.
AbbVie (North Chicago) and Galapagos (Mechelon,
Belgium) recently announced a partnership to pursue
the clinical development of new CFTR correctors and
potentiators. In January 2016, the first duo—GLPG222, a
corrector, and GLPG1837, a potentiator—entered Phase I
clinical studies. Plans are in the works to move two more
candidates—a potentiator called GLPG2451 and the next
generation corrector GLPG2737—into clinical testing by is based upon observations of children with CF, as
the second half of 2016. The testing of double and triple young as four weeks old, who have elevated levels
combinations of this batch of potentials are expected of inflammatory markers which further drive the
to begin in 2017. Preclinical studies indicate that a triple progression of the disease. Corbus’ lead compound,
combination could be effective against up to 90% of Resunab, is an oral anti-inflammatory that works by
CF cases! binding to and activating the CB2 receptor present on
a variety of immune cells—think macrophages, T-cells,
Vertex is also working on next generation CFTR
and B-cells. Activation of the CB2 receptor inhibits the
correctors, with two products in Phase I clinical testing.
immune function of these cells, potentially resulting in a
Plans for a triple-combination drug—using the approved
resolution of the inflammation. Corbus is promoting the
therapies from their prior lineup—are also expected to
drug as a possible treatment for all CF patients, not just
be up to 90% effective for CF cases.
those with a particular type of mutation, meaning more
avenues of attack.
OUTSIDE OF THE CFTR BOX
Spyryx Bioscience (Durham, NC) has identified a tactic
The direct targeting of the CFTR protein is not the
based on an observation that links the regulatory
only approach to CF drug discovery currently being
protein SPLUNC1 and dehydration which leads to thick
entertained in development circles. Vertex is testing
mucus, the hallmark of CF. Simply put, SPLUNC1 helps
an epithelial sodium channel (ENaC) inhibitor (VX-371)
to modulate how much fluid is absorbed into lung cells.
in Phase II trials. In CF, ENaC is often up-regulated—
Spyryx is currently conducting preclinical testing of
meaning a patient makes much more ENaC than those
an inhalable peptide therapeutic ( a peptide is a short
not suffering from the disease. Up-regulation becomes
segment of a protein) based on the SPLUNC1 protein
a problem because ENaC reabsorbs sodium ions, which
that will enhance the absorption of fluid into lung cells,
also results in water reabsorption, contributing to the
relieving the key symptom of CF.
thick mucus that causes the clinical symptoms of CF. By
impeding ENaC, sodium ions are balanced which lessens The leap from managing CF to a bonafide cure has
the chances of severe symptoms from manifesting. turned into an “all in” battle. With more therapies in the
works, the ability to fight CF on all levels is a hopeful step
Corbus Pharmaceuticals (Norwood, MA) is treating
towards shutting down this disease.
CF as a chronic inflammatory disease. The concept

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THE ANTIBIOTIC ECONOMY Life Science Training from Industry Experts

THE STATE OF ANTIBACTERIALS work by inhibiting bacterial cell wall synthesis. The cell
wall is a layer of sugars and amino acids (peptidoglycans)
One of the greatest public health challenges of the 21st
that surround bacterial cell membranes, providing
century is antibiotic resistance, which occurs when a
bacteria with structural support, protection, and a
few bacteria in a given population develop a genetic
filtering mechanism. These new drugs are a synthetic
mutation that enables them to survive—even in the
lipoglycopeptide—a chemical entity similar enough
presence of antibiotics.
to the peptidoglycans so they are easily incorporated
How do bacteria become drug resistant? Suppose a into the cell wall, but different enough so that once
particular antibiotic inhibits an enzyme required for integrated, cell wall synthesis stops. Without a functional
bacterial replication. If one bacterium mutates so the cell wall, bacteria die. Since human cells do not have cell
enzyme has a slightly different shape, the antibiotic is walls, they are not affected by Dalvance and Orbactiv.
no longer effective. The mutated bacterium lives on and
Sivextra (Cubist, Lexington, MA), works by inhibiting
continues to replicate, even as all the others die off. Over
bacterial ribosomes—the enzyme that makes all
time, this resistant strain becomes dominant, spreading
bacterial proteins. Without new protein production, the
from person to person, remaining unchecked and
bacterium is unable to carry out functions essential for
thriving. It is not uncommon for a strain of bacteria to
life and dies as a result.
become resistant to several different antibiotics, giving
rise to the term multi-drug resistant (MDR) bacteria. Avycaz (Actavis, Parsippany, NJ) is used in combination
with a previously approved antibiotic called
One of the most common types of antibiotic-resistant
cephalosporin. Avycaz inhibits the enzyme beta-
bugs is methicillin-resistant Staphylococcus aureus
lactamase, which bacteria secrete in order to break down
(MRSA), which typically causes potential life-threatening
other antibiotics—essentially boosting the potential of
skin infections. In addition to MRSA, other drug-resistant
its paired antibiotic.
microorganisms of urgent concern include Clostridium
difficile (life-threatening diarrhea), Enterobacteriaceae Although these approvals are encouraging, none work by
(bloodstream infections), Neisseria gonorrheoeae entirely novel mechanisms so it is likely that resistance
(severe reproductive complications), Pseudomonas will eventually come about—underscoring the continued
aeruginosa (pneumonia, bloodstream, urinary tract, and need to discover and develop antibiotics.
surgical site infections), and Mycobacterium tuberculosis
(tuberculosis).
IN THE CLINIC
GlaxoSmithKline (London, UK) has a novel antibacterial
Antibiotic resistance is largely caused by antibiotic
compound, Gepotidacin, in Phase II clinical testing.
overuse—the more a bacterial population is exposed,
Gepotidacin is a topoisomerase II inhibitor, meaning it
the greater the probability of mutations. This
inhibits a bacterial enzyme involved in helping bacterial
begs the question: what is the current state of the
DNA to unwind in order to be replicated. Inhibiting
antibiotic economy?
the enzyme impedes bacterial DNA replication.
ON THE MARKET Gepotidacin is being developed in collaboration with
the US government’s Biomedical Advanced Research
The last few years have seen several new antibiotics
and Development Authority/Defense Threat
make it to the market. Three for the treatment of acute
Reduction Agency.
bacterial skin and skin structure infections, often caused
by MRSA, as well as an antibiotic that enhances the Spero Therapeutics (Cambridge, MA) is developing a
effectiveness of its partner antibiotic. way to more effectively target gram-negative bacteria,
or bacteria whose outer membranes contain negatively
Dalvance (Durata Therapeutics, Chicago, IL) and
charged lipopolysaccharides. Lipopolysaccharides
Orbactiv (The Medicines Company, Parsippany, NJ)
are lipid molecules with sugar groups attached, and

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this combination makes it difficult for antibiotics The compound is also highly toxic to bacteria cells, a trait
to penetrate. By developing novel “potentiator” that should also defer resistance by quickly killing off
compounds that interact with and disrupt these target populations before resistance emerges.
lipopolysaccharides, Spero scientists hope to make these Another large and mostly untapped source of potential
bacteria susceptible to antibiotics that already exist. new antibiotics are the microbes of the ocean.
Examples of gram-negative bacteria include strains that Because of significant environmental differences in
cause pneumonia, meningitis, urinary infections, and temperature, salinity, and pressure between the ocean
gastrointestinal problems. and the lab, these microbes can be near impossible to
Motif Bio (London, UK ) has a novel antibiotic, Iclaprim, grow outside of the marine environment. University
in Phase III for acute bacterial skin infections, including of California San Diego scientists are developing
MRSA and MDRSP (multi-drug resistant Streptococcus ways to circumvent this plight through genomics. By
pneumoniae). Iclaprim works by inhibiting an enzyme isolating and sequencing the DNA of marine microbes,
called DHFR, which plays a key role in bacterial researchers identify gene clusters that they predict will
metabolic pathways. code for antibiotic-like compounds. These genes are then
Melinta Therapeutics (Lincolnshire, IL) recently transferred and grown in bacteria. One such compound
completed Phase III clinical studies for MRSA skin is dubbed taromycin A, which impairs the growth of
infections with Baxdela, a compound that belongs to an several types of drug-resistant bacteria, providing a
established class of antibiotics, the quinolones. Baxdela potential platform for future antibiotics.
works by interfering with bacterial DNA replication. Another new approach can be found in the immune
Melinta scientists are also developing novel antibiotics system of the bacteria itself—the often buzzed about
which inhibit the bacterial ribosome—the giant enzyme CRISPR/Cas 9. CRISPR stands for “clustered regularly
that synthesizes proteins. inter-spaced short palindromic repeats.” These are
short DNA sequences that read the same forward and
RESEARCH & DEVELOPMENT backward and are found dispersed throughout the
Antibiotics have traditionally been found by screening genomes of many types of bacteria. In the mid-2000’s,
soil bacterium for compounds they release to kill their scientists realized these sequences were a part of the
competitors: other bacteria competing for the same bacterial immune system. Bacteria insert bits of invading
resources. The big problem with this methodology, viral genomes between the CRISPR sequences. If the
however, lies in the fact that most soil bacteria do not bacteria is subsequently infected by the same virus, the
grow well in the lab—making it impossible to screen 99% previously inserted viral DNA is used to make RNA that
of potential candidates. recognizes and binds to the invading viral genome—
triggering the protein Cas9 to cut up the viral DNA.
A team of scientists at Northeastern University
(Boston, MA) developed a method of cultivating bacteria Researchers at Tel Aviv University and MIT are working
in the lab by sandwiching them between layers of the soil to develop CRISPR/Cas9 systems to target bacteria-
separated by a semi-permeable membrane. NovoBiotic encoded antibiotic resistance genes. Their approach
Pharmaceuticals (Cambridge, MA) used the technique involves encoding CRISPR sequences on either side of an
to screen 50,000 new types of soil bacteria and identified antibiotic resistant gene, which may trigger the gene’s
25 new drug candidates. NovoBiotic’s most promising destruction. The genes can potentially be delivered via
candidate is Teixobactin, a compound that binds to two viruses (called bacteriophages), which in turn can be
different lipid components of the bacterial cell wall, engineered to target specific bacteria. This precision
preventing them from being used to produce future cell technology eliminates the issue of accidentally wiping
walls. Neither lipid is rapidly evolving, so it is likely to out “friendly” bacteria along with harmful bacteria—a
take a much longer time for resistance to come about. problem seen with many current antibiotics.

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A DROP OF GENE THERAPY Life Science Training from Industry Experts

A WALK IN THE PARK FOR HEMOPHILIA? GUNNING FOR GENE THERAPY

Imagine tripping over your feet during a leisurely stroll Hemophilia is a monogenic disease—mutations in only
down the sidewalk. Ouch! Your knees are scraped below one gene can cause the disease—which makes for a
your shorts and blood starts to drip. A quick wipe from a tantalizing therapy target. If scientists can deliver a
conveniently pocketed napkin and soon enough, you are correct copy of the gene that codes for the deficient
the proud owner of some new scabs. Life goes on. clotting factor, then the patient will be able to make the
If you are one of the 20,000 Americans with hemophilia, clotting factor for himself, and no longer be forced to rely
this story has a much different ending. Hemophilia is a on infusions.
condition which prevents scabs from forming due to a Gene therapy relies on viral vectors to deliver genes
lack of functional blood clotting factors. You would need to patients. A virus is simply a segment of genetic
to start replacement therapy right away, or the bleeding material—either RNA or DNA—encapsulated in a protein
could last days, weeks, or even lead to death. coat. Proteins on the surface of the virus help to target
Getting the clotting protein factors for a bleeding it to proteins on the surface of a specific host cell.
hemophiliac involves receiving donor plasma or an Once inside the host cell, certain viruses have evolved
infusion of biologic drugs produced using recombinant to incorporate their genetic material into the host’s
DNA technology. Unfortunately, these traditional genome, in order to “trick” the host cell into making
replacement therapies have been cumbersome in viral proteins.
practice, especially for patients with severe hemophilia. Scientists have been able to adapt these viruses into
Clotting factors are not stable: they have a half-life delivery vehicles for therapeutic genes by tweaking them
of between 10 and 25 hours, so infusions have to be so that they do not cause illness to the person and do
repeated every 48 hours. target the therapeutic tissue of interest. Using standard
Significant improvements in treatment efficacy have genetic engineering techniques, the virus can be further
increased the quality of life for hemophiliacs since modified to contain the therapeutic gene.
our original issue on this topic, which aired in June of
2014. Let’s find out what’s on the market and in the
IN THE CLINIC: GENE THERAPY
pipeline, hopefully making for an easier walk in the park Over a decade ago, gene therapy stalled out in the
for hemophilia. clinic due to safety concerns. Researchers have now
developed vectors that enable better control over
MEN-BER’S ONLY CLUB where the therapeutic gene is inserted into the genome,
There are two types of hemophilia, A and B. Both are making the therapies much safer. As a result, there are
tied to the clotting factor involved in stopping scabs; now several companies with gene therapy treatments
Hemophilia A, the most common, lacks clotting Factor in late-stage clinical trials for hemophilia and other
VIII and hemophilia B is deficient in Factor IX. serious illnesses.

The disease is X-linked: the genes involved are carried BioMarin (Novato, CA) reported encouraging results
on the X chromosome. Since women have two copies of in April 2016 from an ongoing gene therapy trial for
the X chromosome, they are almost never affected by patients with hemophilia A. Factor VIII is normally
hemophilia (you only need one good copy of the gene to produced in the liver, so BioMarin scientists engineered
produce enough clotting factor.) Men only have one copy a liver-targeting virus to deliver the gene for Factor
of the X chromosome, so if the clotting factor gene is VIII. After treatment, six of eight patients produced
mutated, they develop hemophilia. clinically meaningful amounts of the Factor VIII protein.
Although these results are preliminary, and the number
of patients small, they have generated much excitement

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 immune response to the therapeutic virus. By modifying
the virus to be less recognizable by the immune system,
co-administering immunosuppressants, and decreasing
the amount of virus given to patients, the problem might
be solved. However, decreasing the therapeutic dose
limits the potential efficacy. Even for the small dose,
many patients showed an immune reaction against the
treated cells.
Enter the Padua gene variant of Factor IX, named after
the town in Italy where a young man once lived with
severe blood clots. It turned out his version of the Factor
IX gene produced a Factor IX protein with 776 times the
activity level of the normal variant. By using this Padua
gene variant in the viral vector, clinical researchers were
able to deliver much lower doses and still see a clinical
within the gene therapy community because they are the
effect. Spark Therapeutics and Baxalta are both using
first gene therapy trials to meet with any success at all
the Padua variant in their vectors. A life-threatening gene
for hemophilia A.
variation in one may prove to be life-saving for many.
Spark Therapeutics (Philadelphia, PA) has also made
headlines in recent months for its success in gene IN THE CLINIC: GENOME EDITING
therapy trials to treat hemophilia B. In four patients
Editing the genome itself is another approach eyeing the
treated with the viral vector delivering Factor IX, all
clinical trial space. Using zinc-finger nuclease genome
are now producing about 30% of average levels of
editing, Sangamo Biosciences (Richmond, CA) has
the protein—enough to enable patients to go without
successfully corrected a mutated Factor IX gene in mice,
replacement therapy. Despite the small sample
and Sangamo is now conducting Phase I clinical studies
size, the consistency of these results has impressed
with hemophilia B patients. Plans are in the works to file
clinical researchers.
an IND for hemophilia A treatments later this year.
Other companies that have reported preliminary positive
The push for better hemophilia treatments might finally
results for gene therapy trials for hemophilia B include
get the much-needed shove into the market if the
Baxalta (Bannockburn, IL) and Uniqure (Amsterdam,
above-mentioned gene therapies pan out. From 48-hour
Netherlands). Dimension Therapeutics (Cambridge,
infusions to using the power of our genes will make a
MA) started a trial in May 2016.
huge difference in the lives of those suffering from the
disease. The sheer force of gene therapy and genome
THE TALE OF A MAN FROM PADUA
editing are to be reckoned with, and we will be watching
One of the hurdles preventing past hemophilia B gene this space to see where the drops may fall.
therapies from success involved an overly intense

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BREAKING BAD WITH NSCLC & SCLC Life Science Training from Industry Experts

BREAKING BAD it is usually present at very low levels. However, it can

sometimes build up to dangerous levels in well-insulated,
The hit TV series Breaking Bad features anti-hero
tightly-sealed homes built on soil rich in uranium,
Walter White, who starts out as a sympathetic
thorium, or radium.
character: a mild-mannered high school chemistry
teacher with a nagging cough that turns out to be Asbestos used to be a common insulating material
lung cancer. Money problems precipitated by costly used in buildings and ships. The microscopic fibers in
treatments, poor insurance, and a modest salary push asbestos can be inhaled and become lodged in lung
him to start cooking up meth to ensure the financial cells, triggering the activation of inflammatory pathways
security of his family. Spoiler alert: The treatments that result in the release of mutagens and factors that
succeed beyond his expectations, restoring his health promote tumor growth. Since its hazards became
long enough for him to become an unexpected well-documented in the mid-1970s, it is no longer used
meth kingpin. as insulation.

Breaking Bad is a fictionally extreme example of the In addition to carcinogen exposure, there are likely
chaos that can arise from a lung cancer diagnosis. In fact, genetic elements that make certain individuals more
lung cancer is one of the leading causes of cancer-related or less susceptible to lung cancer. Even though 90% of
deaths in the United States. Today’s WEEKLY will give you lung cancer cases are caused by smoking, only about
an update on the causes, types of lung cancer, and the 10% of smokers get lung cancer. In African-American
latest treatments in the clinic and in the marketplace. populations, even when differences in smoking rates and
access to healthcare are controlled for, the rates of lung
THE DANGER cancer are higher. Both of these scenarios suggest that
there may be genetic factors that make certain people
While Walter White did not smoke cigarettes, 90% of
more (or less) susceptible.
those affected by lung cancer are smokers. Other causes
of lung cancer include environmental or workplace
SMALL CELL
exposure to carcinogens (known cancer-causing agents)
such as radon, asbestos, or air pollution. About 10% of lung cancer is small cell, meaning it occurs
in the very small cells found in the bronchii—the tubes
Smoking causes cancer because the inhaled smoke
that branch off of the trachea, enter the lungs, and
contains a range of chemicals, 70 of which are known
divide into even smaller branches within the air sac.
to be carcinogens, including benzene, formaldehyde,
methanol, and acetylene. Some carcinogens are There are currently no targeted therapies available
genotoxic, meaning that they cause cancer by directly for small cell lung cancer, with chemotherapy and/or
interacting with and damaging DNA. If that DNA damage radiation used as the main line of treatment. Broad
occurs in a gene involved in regulating cell division, ranging therapies that harness the immune system are
cancer may result. Non-genotoxic carcinogens have no in the pipeline—Merck’s (Kenilworth, NJ) Keytruda in
direct interaction with DNA, rather they disrupt cellular conjunction with chemotherapy, known as PembroPlus,
structures and change the rate of either cell division or is in Phase II.
processes that increase the rate of genetic error.
NON-SMALL CELL
Radon gas exposure can result in cancer because it is
Cancer that occurs within any cell outside of small cells is
radioactive, and the high-energy radioactive particles
referred to as non-small cell lung cancer (NSCLC), making
given off as the gas decays can cause direct damage to
up the majority (~90%) of lung cancer cases.
cellular DNA. Radon gas is released from the normal
decay of radioactive elements occurring naturally in soil A number of drugs targeting new blood vessel growth—
and rocks. Radon is not considered dangerous because angiogenesis inhibitors—have been approved for the

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treatment of NSCLC. These include Avastin (Genentech;
South San Francisco, CA) and Cyramza (Lilly;
Indianapolis, Indiana).
There are also drugs that target specific NSCLC–
associated mutations. For example, between 10% and
35% of NSCLC cases are caused by the over-expression of
the growth factor receptor EGFR. These types of NSCLC—
more common in non-smokers—can be treated by drugs
that target and inhibit this receptor. These include Iressa
(AstraZeneca; London, UK), Vectibix (Amgen; Thousand
Oaks, CA), Tarceva (Roche; Basel, Switzerland), Afatinib
(Boehringer Ingelheim; Ingelheim, Germany), Portrazza
(Lilly), and Tagrisso (AstraZeneca). Asians are much
more likely than other races to carry an EGFR mutation. the delivery of a highly toxic chemotherapy agent. Not all
NSCLC patients display high levels of DLL3, but for those
About 5% of NSLC cases are caused by mutations in a
who do, Rova-T, which is currently in Phase III clinical
gene known as anaplastic lymphoma kinase (ALK). ALK
testing, could offer new hope.
proteins activate cell division, and mutated versions
can drive cell division inappropriately. The drugs Xalkori Merrimack Pharmaceticals (Cambridge, MA) has a
(Pfizer; New York City, NY), Zykadia (Novartis; Basel, monoclonal antibody, seribantumab, in Phase II clinical
Switzerland), and Alecensa (Roche) inhibit ALK. First development. Seribantumab inhibits activation of the
approved in 2011, Xalkori was granted an expanded HER3 growth factor receptor, and has just been awarded
approval in March 2016 for ROS-1 positive NSCLC. Like Fast Track status by the FDA.
ALK, ROS-1 is an enzyme which activates cell division,
and is mutated in about 2% of lung tumors. COCKTAIL FODDER:
The checkpoint inhibitor drugs Opdivo (Bristol-Myers
WALTER’S DIAGNOSIS
Squibb; New York City, NY) and Keytruda (Merck; Diagnosed with NSCLC, Walter White specifically had an
Kenilworth, New Jersey) have also been approved for inoperable stage 3A adenocarcinoma. This means the
NSCLC patients whose cancers start growing again cancer was initiated in the mucus-producing cells of the
after chemotherapy. lungs and had spread to the lymph nodes (or other sites
near the lungs), but had not spread to distant sites within
Two novel monoclonal antibody therapies are also in
the body. Some types of adenocarcinomas are caused by
development for NSCLC. Rova-T (AbbVie, North Chicago,
ALK mutations, so it is possible that Walter’s miraculous
IL) is an antibody-drug conjugate which targets lung
recovery was caused by one of the ALK inhibitors
cancer cells expressing high levels of a protein called
discussed above.
DLL3 on their surface. Rova-T uses that protein to target

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THE PLIGHT & PROMISE OF P53 Life Science Training from Industry Experts

A HOLY GRAIL? On the left is the structure of p53 bound to DNA’s double
helix. Four domains of the p53 protein bind to DNA in a
What if scientists could identify a mutated gene that
cooperative manner. On the right is an example of one of
accounts for more than half of all cancerous tumors?
the p53 domains bound to DNA shown in light grey. The
This gene would become a holy grail of cancer research.
DNA binding surface of the p53 molecule neatly fits into
This gene exists, and it is called p53. the grooves of the DNA helix.
The name p53 refers to its molecular mass. It is Most of the mutations that occur in cancer—called
sometimes nicknamed “Guardian of the Genome” mutation “hotspots”—happen in the DNA binding
because p53 protects cells from damaging mutations. domains close to the DNA binding surface. These
Mutations in p53 are often associated with various mutations destroy p53’s ability to interact with
cancers for this very reason: if the gene is not doing its DNA. If p53 is no longer able to bind DNA, it can’t
job correctly, the entire genome is more likely to incur trigger cell death in damaged cells. By studying p53
mutations which may affect its ability to regulate cell structural abnormalities, scientists are learning how to
growth and division, potentially leading to cancer. develop new drugs that compensate for the aberrant
Much like Sir Lancelot’s answer to the Bridgekeeper’s protein structures.
question, “What is your quest?”, let’s cross the bridge “to
seek the holy grail.” PROMISING PEPTIDES
A peptide is simply a short (fifty or fewer) sequence of
GUARDIAN OF THE GENOME amino acids. Peptides are often better able to target
p53’s purpose is to trigger cell death—apoptosis— specific protein structures than small molecule drugs
in response to excessive DNA damage. To perform because their shape can mimic naturally occurring
this function, p53 has to bind to DNA in a very protein-protein interactions.
specific manner: Although many p53-associated cancers are caused by
mutations to the p53 gene itself, about half are the
result of p53 inactivation caused by the tight binding of a
second protein, MDM2. Drugs that inhibit or prevent this
MDM2 interaction could potentially result in restoring
full activity to p53, enabling it to carry out its cell-
protecting mission. Aileron Therapeutics (Cambridge,
MA) is working to prevent MDM2 from closely
binding to p53 in their new class of therapeutics—
“stapled peptides.”
Aileron’s stapled peptides have been chemically
modified to be more stable in the patient’s body, and to
enter cells more easily. Their lead drug candidate, ALRN-
6924, binds MDM2 and disrupts its interaction with p53,
freeing p53 up to do its job. ALRN-6924 is in Phase I
clinical studies.
ADRx (Thousand Oaks, CA) is a biotech better known for
focusing on Alzheimer’s disease (AD) rather than cancer.
However, company researchers view one type of p53
mutation similarly to the amyloid-beta plaques linked
to AD. This particular misshapen p53 forms clumps

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reminiscent of amyloid-beta plaques found in AD,
rendering the protein ineffective. ADRx has a peptide
inhibitor of this interaction in preclinical development.

FIXING UP P53
Aprea AB (Stockholm, Sweden) is developing a small
molecule drug, APR-246, which binds to mutated p53 and
restores its correct structure and function. In preclinical
studies, APR-246 showed strong anti-tumor activity
against a range of cancers, including ovarian, small cell
lung cancer, esophageal, and leukemia.
The drug has completed Phase I safety testing in humans
and is preparing to enter Phase II efficacy studies for
ovarian cancers. It is being tested in patients who have COCKTAIL FODDER: AN ELEPHANT-
shown resistance to platinum-based chemotherapy SIZED INSPIRATION
agents such as cisplatin and doxorubin, two drugs Elephants very rarely get cancer. One would expect them
that are normally effective in ovarian cancer. These to get the disease more often than smaller animals due
drugs work by promoting additional damage to cancer to the vast amount of cells in their bodies and the fact
cell DNA, in hopes of triggering apoptosis. However, that they live almost as long as humans. Researchers at
in patients with defective p53, apoptosis does not the University of Utah have pinpointed the likely reason
get triggered. Preliminary clinical data suggests that for their longevity: extra copies of the p53 gene. Humans
APR0-246 helps to overcome resistance to cislatin and have two copies while elephants have 40! By studying
doxirubin. APR-246 is the first compound in clinical elephants and other cancer-resistant animals such as the
development that reactivates mutant p53 inside of naked mole rat, scientists may find additional protective
cancer cells. proteins that could inspire human therapeutics.

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BIOTECH IN SPACE! Life Science Training from Industry Experts

EXPERIMENTING IN LOWER PLENTY OF BONES ABOUT IT

EARTH’S ORBIT The therapeutic area most associated with microgravity
Drug discovery in space? If this sounds like a page from research is osteoporosis, which causes bones to
science fiction, think again. Leading pharmaceutical become weak, brittle, and easily broken. In healthy
companies are now collaborating with NASA to run people, bone is constantly being broken down
experiments on the International Space Station and replaced. When bone is broken down more
(ISS). The focus on bone density, protein structure, quickly than it is replaced, the loss of bone mass
vaccine development, immune function, and aging causes osteoporosis.
brings a swath of potential to this fascinating endeavor. One of the major factors that contributes to healthy
In this issue, we’ll understand the reasons for doing bone mass is weight-bearing exercise, such as walking,
biomedical research just outside the Earth’s orbit jogging, or weight lifting. On Earth, just walking around
and highlight some of the most intriguing projects on every day can help your bones. In contrast, astronauts
the move. on a mission must use exercise machines to ensure they
3... 2... 1... Blast off! do not lose too much bone mass while in space.
Being on the ISS presents a perfect opportunity to
THE GRAVITY OF THE SITUATION improve the preclinical testing of osteoporosis drugs.
First, the obvious question—why? Drug discovery On Earth, it is very difficult—if not impossible—to
research is already extremely resource-intensive on control for differences in an animal subject’s activity
Earth. Why add to the time, effort, and expense by level as influenced by bone mass. In a microgravity
conducting research in space? Because of the unique environment this variable is removed, and investigators
environment offered by the ISS: microgravity. can conclude that any increases in bone mass are likely
As the space station orbits around the earth in free due to an experimental drug.
fall, so too does everything contained inside. The One FDA-approved osteoporosis drug, Prolia, was
result is an environment of weightlessness that can tested in mice on the ISS. Prolia (Amgen; Thousand
be maintained for significant periods of time, enabling Oaks, CA) is a monoclonal antibody that inhibits the
observations and the completion of certain technical cells that break down bone.
tasks unachievable on the ground.
HIGHER QUALITY
PROTEIN CRYSTALLIZATION
Many drugs work by simply inhibiting an overactive
protein. For example, overactive kinase enzymes
can signal cells to divide when they should be at
rest, leading to cancer. It is much easier to develop
an inhibitor against a drug target when the target’s
structure is well characterized. One of the most
effective small molecule kinase inhibitors, Novartis’
(Basel, Switzerland) Gleevec, was developed based on
knowing the structure of the kinase BCR-ABL.
In order to determine protein structure, researchers
must first obtain a crystallized form of the protein,
which in many cases is not possible. Enter microgravity.
Various researchers, largely from academic and

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government labs, have demonstrated that the quality can take years on Earth as compared to months in
and size of protein crystals generated in microgravity is microgravity. These identified genes are then deleted to
often much higher than those made on Earth. make a vaccine strain—a weakened version of the virus
Now, pharmaceutical companies are getting in on the that still produces antigens which are recognized by the
action—Lilly (Indianapolis, IN) is conducting protein immune system, but lacks the ability to make people
crystallization experiments in collaboration with NASA. sick. The company is getting ready to begin clinical
Lilly is also investigating the potential that microgravity testing of a salmonella vaccine initially developed on
eases the process of protein freeze drying and solid- the ISS.
liquid-gas mixture processing, both crucial to biologic
drug formulation. Merck (Kenilworth, NJ) is also running
LONG LIVE THE T-CELL?
protein crystallization studies on the ISS. It’s been known for several years that space travel
has immunosuppressive effects on astronauts. Now,
NEEDLING IN ON VACCINES experiments conducted with T-cells grown on the
Vaccine development is yet another area of medicine ISS point to changes which closely mimic changes in
that has benefited from microgravity research; it turns aging populations. Observations show aging T-cells
out various pathogens increase their virulence more responding slower and less effectively against antigens.
quickly in space than on Earth. Dr. Millie Hughes-Fulford, a former astronaut and
current investigator at the Veterans Affairs Medical
Austin-based Astrogenetix is using microgravity to
Center in San Francisco, is studying these T-cell
better understand pathogens such as salmonella and
changes. Her work may lead to boosting the immune
methicillin resistant streptococcus aureus. Astrogenix
systems of aging populations or other immuno-
was able to identify the genes attributed to both
compromised people in innovative ways.
pathogens’ ability to cause disease—a process that

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WITHOUT A TREG TO STAND ON Life Science Training from Industry Experts

REGULATORY T-CELLS Tregs to tamp down the immune response. TxCell is

MAKE A PATHWAY developing therapeutics based on isolating Tregs from a
patient’s blood, expanding them in the lab in response
After our focus on T-cell activated immunotherapies
to a specific activating antigen, and infusing them back
revving up the immune response, we turn to the other
into the patient. Although the process is expensive, the
side of the coin—a subset of helper T-cells known as
extracted material can be frozen and used as needed,
regulatory T-cells.
providing enough material for five years’ worth of
Regulatory T-cells (Tregs) suppress the immune treatment every few months.
response. The exact mechanism by which this
Their most advanced clinical-stage product is Ova-Treg,
suppression occurs is not entirely understood, but
which is in development for Crohn’s disease. Tregs
some of the effect is due to the release of anti-
are exposed to the protein ovalbumin, which makes
inflammatory cytokines. They are thought to be
up about 60% of egg whites. The Tregs recognizing
activated by “Tregitopes”, or short peptides recognized
ovalbumin are expanded and infused back into the
by Tregs.
patient’s body. Tregs naturally migrate to sites of
Relatively recent arrivals on the drug discovery and major inflammation—or in the case of Crohn’s disease,
development scene, Tregs are quickly attracting the gut. Although ovalbumin is not related to the
the attention of those interested in developing new pathology of Crohn’s, it is typically present in the gut,
pathways to diabetes, cancer, Crohn’s disease and other so the ovalbumin-sensitive Tregs will be activated to
autoimmune disorders. In this WEEKLY, we’ll take a secrete anti-inflammatory signals. Ova-Treg is currently
closer look at some of these innovative applications. in Phase II clinical studies. TxCell believes different
activating antigens can be used to activate Tregs in
IN DEVELOPMENT: TXCELL other parts of the body and is pursuing other avenues
TxCell (Valbonne, France) aims to treat severe chronic in the chronic inflammatory and autoimmune arena.
inflammatory and autoimmune diseases by using TxCell is also working on a second Treg platform—
CAR-Treg. This technology is very similar to CAR-T
therapy discussed in previous issues, only instead
of engineering killer T-cells to recognize targets,
Tregs are engineered to target a specific area of
inflammation. TxCell’s CAR-Tregs are currently in
preclinical development.
Also joining the Treg race is T-cell pioneer Carl June’s
Tmunity Therapeutics (Philadelphia, PA). The company
has announced plans to develop CAR-Tregs as one
of the many immune-based therapies under their
engineered T-cell umbrella.

IN THE CLINIC:
CALADRIUS BIOSCIENCES
Caladrius Biosciences (Basking Ridge, NJ) is conducting
Phase II clinical studies of a Treg-based treatment
for Type 1 diabetes. An autoimmune disorder in
which the immune system attacks insulin-producing
pancreatic cells, Type 1 diabetics find as many as 20%

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of their insulin-producing cells are still intact at the FIGHTING THE TREGS IN CANCER
time of diagnosis. Caladrius’ treatment suggests that
An overabundance of Tregs is a contributing factor to
if the insulin-producing cells can be protected from
the immunosuppressive characteristics of the tumor
destruction and restored to function, the clinical
microenvironment—which means suppressing Tregs
progression of the disease may be halted, making
themselves is a potential avenue for cancer therapy.
control of blood glucose levels more manageable and
By lessening the Tregs in the area around a tumor,
reducing the risk of long-term complications.
the immune system could potentially find it easier to
Caladrius’ Type 1 diabetes product CLBSO3 is a identify and attack cancer.
collection and expansion of the patient’s own Tregs. The
Tizona Therapeutics (South San Francisco, CA)
company began Phase II clinical testing in March 2016.
is developing a monoclonal antibody therapeutic
Enrolled patients receive a single administration of
targeting Tregs with the goal of depleting them from the
Tregs and will be monitored for two years.
tumor microenvironment.
IN DEVELOPMENT: TREGITOPES AbbVie (North Chicago, IL), in partnership with Argenx
EpiVax (Providence, RI) is bypassing cell therapy (Rotterdam, Netherlands), recently announced a
altogether by relying on “Tregitopes”—short amino program to develop an inhibitor of GARP—a protein on
acid sequences that activate subsets of Tregs. The idea the surface of Tregs that enables the production of anti-
is to administer Tregitopes together with an antigenic inflammatory signaling molecules. By inhibiting GARP,
protein, in hopes of activating antigen-specific Tregs Tregs should be disabled, allowing cancer to be targeted
within the patient. Currently conducting preclinical and taken down.
studies in Type 1 diabetes and various allergies, Flexus (San Carlos, CA) is an early-stage biotech
EpiVax anticipates their approach may be useful in company developing small molecule inhibitors of
preventing immune reactions to a range of therapeutic Tregs. Recently acquired by Bristol-Myers Squibb
interventions such as therapeutic proteins, organ (New York, NY), their pipeline is attracting additional
transplants and gene therapies. excitement and attention to Tregs as the next big thing
in immunotherapy.

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ON A TUMOR’S TURF Life Science Training from Industry Experts

SOLID TUMORS DEFINED

A solid tumor is defined as an abnormal mass of tissue
that usually does not contain cysts or liquid areas
according to the National Cancer Institute.
Solid tumors may be classified as:
• Benign (noncancerous) if they do not invade nearby
tissue or spread to other parts of the body.
• Malignant (cancerous) if they invade and destroy
nearby tissue and spread to other parts of the body.

THE TUMOR MICROENVIRONMENT

The tumor microenvironment (TME) describes the
cellular environment in which the tumor exists.
Different than healthy tissue, the TME may contribute to
tumor growth and complicate treatment strategies.
The TME is typically immunosuppressive—that is, it
actively attempts to prevent the immune system from
recognizing a tumor as harmful. One of the factors
contributing to immunosuppression is the presence
of “suppressor cells”— white blood cells that subdue
rather than activate the immune system. These
include tongue-twister names such as myeloid-derived
suppressor cells, tumor-associated macrophages, and
TACKLING THE SPACE regulatory T-cells.
AROUND SOLID TUMORS Suppressor cells release signaling molecules that “turn
Covering the science behind T-cell-based down” activated immune cells within the immediate
immunotherapies has been the name of the game for area. They also release growth factors that increase
our past couple of issues. CAR-T and TCR therapies blood vessel growth into the tumor, which helps the
show significant promise in early phase blood cancer tumor to survive. The TME also inhibits the display
clinical trials, but what about solid tumors? of antigens (tumor-specific proteins) on the tumor
surface, which are necessary to activate killer T-cells
Previously mentioned Juno Therapeutics’ (Seattle,
against cancer.
WA) Armored CAR technology has declared war on
tumors, and as you will read below, there are plenty of Harnessing the space around the tumor might
interesting battle grounds to choose from. Let’s find herald some fighters in the field. Surface Oncology
out why solid tumors are more difficult to treat than (Cambridge, MA) is using a combination of proprietary
leukemia, lymphoma, and myeloma and where the technologies to modify the TME. These modifications
biopharma industry is staging its fight. include blocking suppressor cells, stopping
immunosuppressive signaling molecules, and improving
the presentation of antigens on tumor cell surfaces.
Currently in the preclinical stage, Surface recently
partnered with Novartis (Basel, Switzerland) to

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further develop their platforms. Clinical development currently conducting early stage clinical trials using
candidates are expected to be released in the engineered Listeria monocytogenes against pancreatic
near future. cancer, non-small cell lung cancer, ovarian cancer,
and mesothelioma.
BACTERIA TO THE RESCUE?
In the context of immunotherapies, most people use THE STING PATHWAY
the term “cell therapy” to refer to the activation or Aduro Biotech is also working on a small molecule
engineering of T-cells to attack cancer. Aduro Biotech immune system activator to help overcome the
(Berkeley, CA) puts a new spin on the concept by immunosuppressive TME. These small molecules are
using engineered bacteria cells to induce a cancer- modified versions of naturally occurring molecules
specific immune response—the technology aims to get that activate a pathway known as STING: Stimulator of
around the problem of poor antigen presentation. The INterferon Gene.
concept: to uncloak tumor cells so they are more easily Activation of the STING pathway results in the
identifiable by the immune system. production of immune-activating signaling molecules.
Scientists at Aduro have taken the pathogenic Preclinical studies suggest that delivering the STING-
bacterium Listeria monocytogenes and modified it in activating molecules near tumor sites invokes
two ways: the immune system to recognize and respond to
• Safety modification: Two genes that are critical to its cancer cells.
ability to infect liver cells were deleted. If these possibilities become a reality, the battleground
• Efficacy modification: Two genes that code for around a solid tumor may be taken by the world of drug
tumor-specific antigens were added. discovery, adding to the arsenal of immunotherapies in
our fight against cancer.
When delivered to a patient, these cells should
activate tumor-specific T-cells to attack. Aduro is

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NEXT-GENERATION CAR-T Life Science Training from Industry Experts

THE RACE IS ON In ACTR, the antibody part of the cell attaches to a

monoclonal antibody (mAb) delivered separately to the
Our last WEEKLY—Hacking the Immune Response—
patient. (Recall in CAR-T the antibody part of the cell
unveiled the science behind CAR-T and TCR, two
attaches to the cancer cell directly.) First the mAb locks
immunotherapies under the microscope of the
onto the cancer cell and then ACTR locks onto the mAb.
mainstream press. The well-deserved media attention
highlights the ability of these “living drugs” to recognize
and obliterate cancers.
With all of the early phase clinical success, a few
challenges have popped up:
• S
 afety: CAR-T and TCR can set off an immune
response that is too strong, which could result in
life-threatening adverse reactions.
• C
 ost: The high price of engineering CAR-T
and TCR therapies for each patient is an
exceedingly expensive.
As we know, the biopharma industry loves a challenge.
In this WEEKLY, we’ll continue our CAR-T focus by
examining next generation T-cells that may prove to be ACTR has two key advantages:
safer, more effective and more cost conscientious. • T
 he same ACTR cell type could be used to target
different cancers—the physician would simply need
ACTR COST ADVANTAGE to change the monoclonal antibody biologic used in
Unum Therapeutics (Cambridge, MA) is working on a conjunction with ACTR. Producing ACTR in bulk may
next generation T-cell, referred to as antibody-coupled keep costs down.
T-cell receptor (ACTR). • A
 dverse reactions could be halted when the patient
stops taking the mAb, because discontinuation of
the mAb essentially acts as an “off” switch in cases
where the immune response is too strong. The
combination of ACTR and mAb could increase the
safety of the T-cell therapy.
Unum has one ACTR—used in combination with non-
Hodgkin’s lymphoma, targeting Rituxan (Genentech;
South San Francisco, CA)—in Phase I clinical trials and
several more in preclinical development.

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BISPECIFIC CAR SAFETY antibodies. This triggers a release of cytokines,
which revs up the immune system. Cytokines may be
Juno Therapeutics (Seattle, WA) is developing more
particularly valuable in solid tumors, which are often
ways to control CAR-T. One approach is bispecific
harder to treat than the blood cancers which have seen
CAR-T: engineered T-cells with two different chimeric
early clinical success with first-generation CAR-T.
antigen receptors.
Juno has one armored CAR in Phase I clinical studies
The goal is to increase specificity for cancer
and several more in preclinical development.
cells and decrease specificity for healthy cells.
Two considerations: OFF-THE-SHELF CAR-T COST SAVINGS
• E
 ngineer a bispecific CAR-T to recognize not First generation CAR-T has been patient-specific, or
one, but two cancer-specific proteins to be autogenic. In other words, T-cells are first removed
fully activated. from the patient to be treated, modified, and infused
• E
 ngineer a bispecific CAR-T to activate in the back in. Customizing CAR-T in this manner is time
presence of one cancer-specific protein, but if a consuming and expensive but necessary to avoid
second protein is also recognized, an inhibitory immune rejection.
signal is sent, and CAR-T shuts down. In this case, A few different companies are exploring ways to create
the second protein would be present on the surface “off the shelf” T-cell therapies that can be used with
of healthy cells and not cancerous cells. any patient. The trick is to remove the tell tail signs that
Juno’s bispecific CAR-Ts are in preclinical development. allow the recipient’s immune system to recognize the
engineered T-cell as foreign. Strategies in preclinical
ARMORED CAR EFFICACY development use genome editing to remove the genes
While keeping the immune response under control is responsible for immune rejection. Companies working
important for safety reasons, in some cases, amplifying on developing this type of allogenic (from another
T-cell response may be required for efficacy. Juno’s person) cell therapy include Cellectis (New York, NY),
“armored CAR” technology uses ACTR technology Celyad (Boston, MA), and Adaptimmune (Philadelphia,
(described above) with an increased dose of monoclonal PA).

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HACKING THE IMMUNE RESPONSE Life Science Training from Industry Experts

T-cells roam the body on the look-out for signs of

invasion by recognizing foreign proteins displayed
on virus-infected cells and bacteria. A T-cell is highly
specialized and able to match its own distinctly shaped
receptor to the foreign protein─much like a lock and key
interface. Once a match is made, the T-cell is activated.
Activated T-cells divide rapidly and produce two types
of cells: killer T-cells and helper T-cells. Killer T-cells
trigger cell death and helper T-cells secrete signaling
molecules (cytokines) that rev up the immune response.

TRAINING T-CELLS TO GO
AFTER CANCER
T-cells don’t naturally recognize tumor-specific proteins
on the surface of cancer cells due to the cloaking ability
BIOPHARMA’S DARLING
of cancer. Tumor cells are seen as “self” by our immune
Immunotherapy is the current biopharma darling, system, which partly explains the success of cancer as
garnering massive investment and media attention. a disease.
Ranging from monoclonal antibodies to engineered
To train T-cells to go after a tumor, killer T-cells with
T-cells, companies are rapidly learning how to harness
special markers are first isolated from a blood sample.
the power of the immune system to fight disease.
These selected killer T-cells are genetically altered to
There are two general categories of immunotherapies: create recognition receptors unique to the patient’s
• Activation immunotherapies “turn on” or “turn tumor. These engineered T-cells are then multiplied in
up” the patient’s own immune response to help the lab, and once infused back inside the patient’s body,
fight disease. they seek out and destroy the cancer.

• Suppression immunotherapies suppress the

EASILY CONFUSED: CAR-T VS. TCR
patient’s immune response in disorders caused
by an overactive immune system—think There are two types of engineered T-cell therapies:
autoimmune disorders. chimeric antigen receptor therapy (CAR-T) and T-cell
receptor therapy (TCR).
In this WEEKLY we concentrate our efforts on two
types of activation immunotherapies, chimeric antigen The word “chimeric” means to be composed of parts
receptor therapy (CAR-T) and T-cell receptor therapy from two or more different sources. A CAR-T is part
(TCR). Both rely on engineering T-cells to recognize antibody and part T-cell. The hybrid produces a
specific cancer targets and lay the groundwork for some seek (the antibody part) and destroy (the T-cell part)
of the hottest medicine in development today. molecule that goes after cancer.
Once a T-cell has locked onto its target, it makes copies
WHAT ARE T-CELLS? of itself and sends out cytokines to call the rest of the
Your body is an amazing machine, and just like all immune system to the fight. The killing ability of CAR-
well-designed mechanics it has a built in degree of Ts are extended past the original cell line to the next
redundancy. Should your non-specific immunity fail, the and onward, ideally attacking any and all cancer cells it
immune system will call in the big guns: T-cells. can find.

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 Simply put, CAR-T attacks cancers that display tumor-
specific proteins on the outside of cells and TCR attack
cancers that display tumor-specific proteins on the
inside of cancer cells.

SAFETY CONCERN: CYTOKINE STORMS

CAR-T and TCR are showing great promise (especially
treating blood cancers) in early, small-scale clinical
trials. However, one of the main clinical concerns is
their ability to induce a “cytokine storm.” Activated
T-cells release signaling molecules called cytokines
that stimulate the immune response, but if too many
cytokines are released, a patient can suffer life-
threatening effects. In order for CAR-T and TCR to
become routine clinical practice, fail-safe methods for
CAR-Ts can only recognize proteins expressed on the
controlling cytokine storms are needed.
surface of the cancer cell, but TCRs can recognize
tumor-specific proteins on the inside. Inside tumor-
CONTROLLING THE STORM
specific proteins are broken into fragments; these are
“presented” on the cell surface with another protein One approach to controlling an overactive immune
called MHCI. response is to require two inputs: a target-recognition
domain and a small molecule drug. Rather than having
TCRs are engineered to recognize a tumor-specific
the target-recognition domain directly connected
fragment/MHCI complex. The gene sequences
to the activation domain, they are each controlled
introduced into engineered TCRs are selected by
by a separate “switch” that is “turned on” by a small
screening healthy donors for naturally-occurring TCRs
molecule drug. Once a patient is infused with the T-cell
that recognize a specific fragment/MHC combination.
therapy, the patient is given a small molecule drug
to fully activate the drug. If this results in a cytokine
storm or other overpowering response, the drug can be
immediately withdrawn.
Bellicum Pharmaceuticals (Houston, Texas), Intrexon
(Germantown, MD), and Theravectys (Villejuif, France)
are all working on the preclinical development of
control switches for CAR-T cells.
Next week, we will continue our discussion of T-cell
innovations including the development of “off the shelf”
CAR-T therapy.

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PUSHING THE SELF-DESTRUCT BUTTON Life Science Training from Industry Experts

undergoing treatment have this chromosomal

abnormality. This mutation is identified by Abbott’s
(North Chicago, IL) companion diagnostic Vysis CLL FISH
probe kit, which the FDA approved in 2011.

MECHANISM OF ACTION: VENCLEXTA

Venclexta is a small molecule inhibitor of BCL2, a
protein that inhibits apoptosis. Apoptosis is known
as “cell suicide,” it is initiated within cells that have
incurred a significant amount of DNA or cellular
damage. In healthy cells, BCL2 can stop cell suicide from
happening inappropriately. But in the 17p deletion type
of leukemia, BCL2 is overexpressed, preventing those
cells from initiating apoptosis even when damaged.
PUSHING THE SELF-DESTRUCT BUTTON
Inhibiting BCL2 enables those apoptotic pathways to
Breakthrough drug Venclexta charged onto the be activated, causing the cancer cells to flip the self-
marketplace three months early to battle chronic destruct switch. Venclexta is the first BCL2 inhibitor to
lymphocytic leukemia. AbbVie’s (North Chicago, IL) and be approved by the FDA.
Roche’s (Basel, Switzerland) new therapy gained a quick
approval after 80% of patients in the 106-person clinical THE PLAYING FIELD
trial responded to the small molecule inhibitor.
Prior to Venclexta’s approval, monoclonal antibodies
Chronic lymphocytic leukemia (CLL) is the most such as Rituxan (Roche) and Campath (Sanofi; Paris,
common type of leukemia in adults, with approximately France) targeted proteins found on the surface of
15,000 new cases diagnosed each year according to the B-cells to fight CLL. Small molecule drugs also on the
National Cancer Institute. Let’s find out why the FDA market included Imbruvica ( Janssen Pharmaceuticals;
awarded a Breakthrough Therapy Designation and Beerse, Belgium) and Zydelig (Gilead; Foster City, CA)—
discover what exactly causes Venclexta to push the self- both are inhibitors of enzymes that drive forward cell
destruct button. division inside of cancer cells.

17P DELETION A LITTLE TOO GOOD

The term “leukemia” simply means an overproduction We don’t usually think a highly effective drug can cause
of white blood cells that do little more than continue to a problem. However, sometimes potent cancer drugs
divide. CLL ultimately crowds out healthy cells due to cause “tumor lysis syndrome,” wherein tumor cells die
the uncontrolled growth and accumulation of B-cells in at such a high rate that the release of their contents
the bone marrow and blood. upon lysis (breaking open) can cause changes in blood
Venclexta is specifically approved for CLL patients electrolytes and metabolites. These changes can be
who have a deletion in chromosome 17, known as dangerous and even fatal. During initial clinical trials,
17p deletion; this deletion typically correlates with a Venclexta was so effective that some patients showed
poor prognosis. Today, approximately 10% of newly signs of tumor lysis syndrome, which necessitated a
diagnosed individuals and 20% of those already lower dose as the trials moved forward.

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ATTACK OF THE MIGRAINE Life Science Training from Industry Experts

NEW AVENUES FOR face and is considered a pain-associated pathway in

MIGRAINE THERAPY migraine attacks.

Migraines. One of the most convenient excuses to • Lowered Serotonin Levels: Another area of active
get out of work or dinner with your mother-in-law. research involves the neurotransmitter serotonin.
More than just a headache, migraines often include Serotonin is most often associated with mood—
symptoms like intense pain, nausea, dizziness, and antidepressants such as Pfizer’s (New York, NY)
extreme sensitivity to light or noise. They can last Prozac, which increases levels of serotonin in the
anywhere from a few hours to days on end. Episodes brain. This neurotransmitter is also implicated in
may be as frequent as several times a month, or as migraine pain pathways, with levels dropping during
infrequent as a few times a year. an attack.

As many as 36 million Americans suffer from • Hormonal Link: Migraines are more common in
migraines—about 12% of the population. Although women than men, so a hormonal link may be tied
there are products on the market to treat symptoms into the causality. There is often a reduction in
once started, nothing is currently available to prevent symptoms after menopause.
pain onset in frequent sufferers. In this WEEKLY, we’ll • Glutamate Accumulation: Migraines have
make no excuses as we explore current treatments and been observed to run in families, pointing to
unearth the exciting clinical pipeline. possibilities in the gene pool. In recent years, a
few gene variations that appear to increase the
ATTACK OF THE MIGRAINE risk of developing migraines have been identified
There are three distinct parts of a migraine episode as through genome-wide association studies. Two
outlined by science, however, not all migraine sufferers of these genes result in increased levels of the
experience these phases with the same intensity. Each neurotransmitter glutamate, suggesting that
set of symptoms is unique to the individual, and can accumulation of glutamate in synapses may be
include prodome, aura, or postdome phases. a trigger. A third gene, casein kinase 1 delta
(CSNK1D), produces an enzyme that modifies many
• Prodome occurs in the hours or days before a
different proteins in the brain. Because CSNK1D
migraine attack. It includes mood disturbances, stiff
interacts with multiple targets, it is less clear
muscles, and sensitivity to smells or noise.
how mutations in the gene may be contributing
• Aura is the period just before the severe pain to migraines, but the finding offers support for
attacks. Visual distortions are the most common the idea that there is at least some genetic basis
symptom, with sensory or motor disturbances for migraines.
potentially occurring as well.
• Postdome happens after the actual headache and IN YOUR MEDICINE CABINET
includes symptoms such as lingering pain and Currently, there are no treatments that prevent the
cognitive difficulties. onset of a migraine, but there are medicines that can
ease symptoms and stop the progression of an episode.
THE SCIENCE BEHIND THE EPISODE Some sufferers gain pain relief from over the counter
While the exact cause is largely unknown, there are a non-steroidal anti-inflammatory medicines such as
few theories: ibuprofen or aspirin. If these are not effective, the next
step is to try prescription drugs designed specifically for
• Brain Stem Changes: Research by the Mayo Clinic
the treatment of migraine headache.
suggests migraines derive from changes in the
brain stem and its interaction with the trigeminal Triptans work by binding to and activating serotonin
nerve. The trigeminal nerve supplies feeling to the receptors in the network of blood vessels supplying

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the brain. Triptans are agonists, so the blood
vessels constrict in response to the activation,
which then inhibits the release of pro-inflammatory
neurotransmitters. The swelling in blood vessels goes
down, relieving migraine pain in about 70-80% of
patients. However, triptan use is contraindicated in
those who have a history of cardiovascular disease.

IN THE CLINIC
The hot new kid on the block is calcitonin gene related
peptide (CGRP) inhibitors. CGRP spikes during migraine
attacks, and is thought to play a role in the brain
pathways that process pain. The exact molecular
mechanism of how the CGRP spike is related to migraine from getting to its intended target by blocking as many
onset is not yet fully understood, but that hasn’t entry points as possible. Lilly’s mAb is in Phase III
stopped several leading drug companies from moving testing; Teva’s recently completed Phase II.
forward with clinical testing. Results so far suggest
On the tail end of the migraine development train
that while they may not be a panacea for everyone,
is Allergan (Dublin, Ireland) with its small molecule
CGRP inhibitors may significantly reduce the number of
inhibitor of the CGRP receptor—clinical trials are
migraine days per month in some patients, restoring a
expected to begin mid-2016.
quality of life that could only be dreamed of before.
Alder Pharmaceuticals (South Bothell, WA) and Lilly COCKTAIL FODDER: A
(Indianapolis, IN) both have mAbs directed at the CGRP HEADACHE OF THE PAST
molecule in Phase III clinical testing. The idea is for the
We sometimes blame migraines on the stress of
mAb to “mop up” CGRP before it reaches the receptor
modern living. In some cases, stress is a trigger, but
playing a role in migraine initiation.
the headaches are hardly a modern phenomenon.
Teva Pharmaceuticals (Petah Tikva, Israel) and Descriptions consistent with migraines are found in the
Lilly both have mAbs targeting the CGRP receptor. By ancient Egyptian medical text Ebers Papyrus, dating
selecting for an antibody that binds to yet doesn’t from 1550 B.C., as well as Hippocratic texts dating from
activate the receptor, researchers can prevent CGRP 200 BC.

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DISRUPTING THE CYCLE Life Science Training from Industry Experts

proteins interact with each other and regulate levels

of expression of other proteins throughout an
approximately 24-hour period. This is why most people
feel disoriented after traveling through several time
zones, even if they get an adequate amount of sleep.
These tiny molecular clocks are signaling that it is later
(or earlier) than the local time—and a whole range
of physiological functions from body temperature
to heart rate, blood pressure, and alertness levels
respond accordingly.
Clock proteins are regulated by a “master clock” in the
brain. Technically referred to as the suprachiasmatic
nucleus, this master clock consists of about 20,000
CIRCADIAN RHYTHMS GOVERN MORE nerve cells just above the region where the optic
THAN 24 HOURS OF YOUR DAY nerve crosses into the brain. This location, called the
optic chiasm, explains why many of these neurons are
Overnight shift workers, students pulling an all-
sensitive to light.
nighter to cram for a final exam, and business people
rushing between time zones all share one thing in
ON THE MARKET: HETLIOZ
common: significant disruption to their circadian
rhythm. This roughly 24-hour activity cycle responds People who are totally blind cannot receive the proper
primarily to light and darkness and is found in most light input to control their master clock. As a result,
living organisms—people, plants, animals, and even many suffer from non-24-hour sleep-wake disorder. It
some microbes. becomes virtually impossible to fall asleep at standard
times, which severely impacts their ability to function
Disruption in the cycle can cause more serious
professionally and socially.
consequences on top of a few days of disorientation.
Abnormal circadian rhythms are correlated with In July 2014, the FDA approved Vanda
insomnia, diabetes, increased cardiovascular events, Pharmaceuticals’ (Washington, DC) Hetlioz for patients
some types of cancer, Parkinson’s disease, and more. with non-24-hour sleep-wake disorder. Hetlioz is an
agonist; it binds directly to and activates the melatonin
In this issue, we’ll take a closer look at how circadian
receptor. Melatonin is a sleep-inducing hormone with
rhythms are regulated, examine some links with various
levels increasing at the onset of darkness, linking it to
disease states, and find out how drug discovery efforts
the sleep-wake cycle. Since melatonin production is
are taking this important phenomenon into account.
dysregulated in totally blind people, Hetlioz aims to
normalize sleep patterns. Although melatonin itself
PROTEINS ON THE CLOCK is available in pill or liquid form over the counter,
The circadian rhythm is regulated by a combination synthetic melatonin receptor activators such as Hetlioz
of environmental and internal cues. Environmental are designed to be more stable.
cues are based on light and dark, while internal cues
are simply “biological clocks.” These biological clocks IN DEVELOPMENT: RESET YOUR PRESET
are groups of interacting proteins in cells throughout
Aptly-named Reset Therapeutics (South San
the body.
Franscisco, CA) is focusing on circadian-rhythm
The most extensively characterized biological clocks disruptions as they relate to a whole range of diseases
are appropriately dubbed Clock proteins. The Clock including type 2 diabetes, Cushing’s syndrome

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(elevated cortisol levels), high blood pressure, obesity, medications, however, the time of day the pill is
sleep apnea, cancer, and inflammation. Scientists at swallowed may influence drug efficacy.
Reset have identified potential circadian-modulating In most healthy people, blood pressure is tied to
compounds and are testing them in animal models of circadian rhythm, naturally dropping between 12:00
Cushing’s syndrome and type 2 diabetes. AM and 3:00 AM. In people with high blood pressure,
however, this drop often doesn’t occur naturally. By
PRECLINCAL: SHUTTING DOWN taking blood pressure medication at night, patients can
THE CANCER CLOCK restore this normal decrease in pressure.
A possible link between circadian rhythm dysfunction
Dosage timing may also be important for certain types
and cancer has caught the eye of drug development.
of cancer drugs. For example, PARP1 inhibitors work
One of the functions of the Clock proteins is to set
by shutting down a DNA repair enzyme in cancer
restrictions on when cells can divide, so circadian
cells. The shutdown results in so much DNA damage
disruptions may affect cancer-related cell division.
to the cells that they initiate a cell-suicide program
Researchers at the University of California, Santa
known as apoptosis—the cancer cells pretty much end
Cruz are zeroing in on a protein known as PASD1, which
up killing themselves! Researchers at University of
is expressed in many different cancer cell lines. PASD1
North Carolina (Chapel Hill, NC) have discovered that
interacts with the Clock proteins, essentially interfering
DNA repair enzymes are more active later in the day,
with their function and shutting them down. Early
translating into PARP1 inhibitors possibly being more
preclinical work suggests that the inhibition of PASD1
effective if given in the morning; inhibiting the enzymes
causes the Clock proteins to reactivate—making PASD1
should be easier when they are not at their peak
a possible drug target.
activity level.

TIMING IS EVERYTHING Circadian rhythms govern more than the 24 hours of

your day, and uncovering various mechanisms at the
Taking medicine at the same time every day is a good
cellular level of the sleep-wake cycle might just open up
way to make sure a dose isn’t forgotten. For certain
new avenues to treat a whole array of diseases.

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AN INBORN ERROR OF METABOLISM Life Science Training from Industry Experts

DIET SODA’S OMINOUS WARNING DECIPHERING THE CAUSE

“Phenylketonurics—contains phenylalanine” is listed The exact mechanism behind PKU is not known, but
on many diet sodas, including Diet Coke and Diet we do know the manifestations of the disease─brain
Pepsi. There is no elaboration, just an ominous warning imaging studies show damage to both the white matter
in bold typeface. Since a few of us at BioTech Primer (myelinated axons) and the grey matter (neuronal
are big fans of diet soda, we wondered: Should we be cell bodies).
worried? And what exactly is a phenylketonuric? There are two different disease cascades:
A quick Google search reveals phenylketonurics are • Classical PKU is caused by several possible
people born with the rare disease phenylketonuria mutations in the gene coding for the enzyme
(PKU); their bodies are unable to break down the amino phenylalanine hydroxylase (PAH), which is
acid phenylalanine. In today’s BioTech Primer WEEKLY responsible for breaking down phenylalanine.
we’ll examine the story behind PKU as we continue our Different mutations affect the level of impairment—
series on neurological diseases. some patients lack all ability to metabolize
phenylalanine while others have reduced capability.
PKU EXPLAINED
• PKU is also the result of a deficiency in cofactor
We know from diet soda labels that phenylketonurics
tetrahydrobiopterin (THB). Cofactors are chemical
must specifically avoid the artificial sweetener
compounds (i.e. non-proteins not coded for by
aspartame—think brand names like Equal or
genes) required for the PAH enzyme’s activity. THB
NutraSweet. But an even bigger challenge is avoiding
is essential for the proper functioning of PAH
all high-protein foods—meat, dairy, fish, nuts, beans,
legumes, and soy-based foods such as tofu─which all ON THE MARKET
contain phenylalanine. As many readers know, living
Kuvan is the only treatment for PKU approved by the
with dietary restrictions is no walk in the park, but
FDA. Developed by BioMarin Pharmaceuticals (San
living with one that can cause serious brain damage
Rafael, CA), it is the synthetic form of THB. As a small
is devastating.
molecule drug, Kuvan is taken orally and is effective
There are an estimated 50,000 phenylketonurics under at reducing blood phenylalanine levels in 30% to 50%
the age of 40 living in developed countries. The disease of patients. Indicated for phenylketonurics whose
is autosomal recessive—meaning phenylketonurics disease is primarily the result of THB or for certain
receive two defective copies of the gene. Since healthy PAH mutations resulting in a sub-optimal THB enzyme
parents might be in the dark about their PKU carrier functionality, these patients benefit the most from
status stringent newborn testing is routine. Starting Kuvan, however, a restrictive diet must still be followed.
from day one any infant that tests positive for PKU
must follow a highly restrictive diet because high levels IN DEVELOPMENT
of phenylalanine can potentially cause developmental
BioMarin is currently conducting Phase III clinical
delays, seizures, irregular motor functioning, and
studies of Pegvaliase, an enzyme replacement therapy
mental retardation.
for PKU. This injectable is a recombinant version of
PKU diets in infancy focus on phenylalanine-free baby phenylalanine ammonia lyase, an enzyme related
formulas. Pasta, fruit, vegetables, as well as low protein to PAH, which also breaks down phenylalanine.
bread are recommended when solid foods come into Preliminary study results suggest Pegvaliase reduces
play. Lifelong nutritional supplements are key since the phenylalanine blood levels by 62% when compared
overall PKU diet lacks protein. to placebo, offering hope to those not responding
to Kuvan.

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Rubius Therapeutics (Cambridge, MA) has announced
plans to use its novel red blood cell therapeutics
platform to tackle PKU. Bone marrow stem cells,
marked for red blood cell formation, are programmed
to produce an enzyme that breaks down phenylalanine.
These phenylalanine-metabolizing red blood cells
are grown in the lab and transfused into the patient
where they remain in circulation for several months.
This platform is still in preclinical testing but has
generated much excitement within research and
investment communities.
Synlogic (Cambridge, MA) is also taking a novel
platform approach to this orphan disease. Dubbed
“synthetic biotics,” they plan to engineer gut
microbes—so-called “healthy bacteria”—to produce
phenylalanine-digesting enzymes. Synlogic’s goal is to COCKTAIL FODDER: TOXIC FUNGUS?
create bacteria that can sense changes in phenylalanine Why does the defective PAH gene persist in human
concentrations and respond appropriately. Ultimately, populations, when its effects can be devastating to
the hope is to create a “living drug” that can be taken human health? It’s thought that heterozygotes—
orally and integrated into the patient’s existing gut individuals who have one functional copy of the
microbiome. Synologic’s PKU treatment is currently in gene and one dysfunctional copy—have some
preclinical testing. protection against the toxic effects of the fungal toxin
ochratoxin A, which can occur in grain, pork, coffee,
and grapes. Turns out, having one copy of the gene
may have conferred a survival advantage throughout
human evolution.

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TWO MONOCLONAL ANTIBODIES
WALK INTO THE MARKET Life Science Training from Industry Experts

ATTACKING ASTHMA & Inflammatory cytokines activate the immune response;

PUSHING OUT PSORIASIS anti-inflammatory cytokines turn that response down.
If a person’s body is producing too many cytokines that
Two new monoclonal antibodies debuted on the market
jump start too many immune cells, the immune cells
thanks to FDA approvals last month. Both treat chronic
themselves may also become overactive. The challenge
inflammatory conditions: Lilly’s (Indianapolis, IN)
for drug discovery is figuring out where to intervene to
Taltz for psoriasis and Teva’s (Petah Tikva, Israel) drug
calm an overactive immune response—the nuances of
Cinqair for severe asthma.
biological processes at the cellular level are hardly black
Interestingly enough, having psoriasis is associated with and white. Cell signaling pathways are all shades of
an increased risk of developing asthma according to grey, highly complex with many intermediate steps.
a study in the British Journal of Dermatology. While
these two conditions have different clinical manifesta- EASILY CONFUSED: INTERLEUKIN
tions—constricted airways in asthma vs. red, scaly, itchy & IMMUNOGLOBULIN
skin lesions in psoriasis—both are the result of chronic Interleukins are a particular type of cytokine produced
inflammation often triggered by overactive cell sig- by white blood cells. There are dozens of different
naling pathways inciting the immune response. In this interleukins, each with a slightly different role in tuning
WEEKLY, we’ll take a closer look at the molecular path- the immune system. Interleukins may be used to
ways and various medications designed to control these therapeutically activate the immune system. In chronic
two conditions that can sometimes coexist. inflammatory disorders such as psoriasis, for example,
the strategy is to inhibit the interleukins inducing an
TERM OF THE WEEK: INFLAMMATION overactive immune response.
Inflammation refers to the accumulation of fluid,
Immunoglobulin (Ig) is another term for antibody.
plasma, proteins, and white blood cells initiated by
Although all antibodies have a similar structure, there
a physical injury, infection, or immune response.
are five distinct classes of Ig proteins. Subtle differences
When provoked for the right reasons, inflammation
in structure between these classes differentiate their
is a good thing—it removes damaged cells, irritants,
functions and localization within the body. For example,
and pathogens. In inflammatory disorders such as
immunoglobulin E (IgE) proteins bind to allergens and
psoriasis and asthma, however, the immune response is
trigger histamine release. This is why inhibiting IgE may
inappropriately set off against the body’s own healthy
be useful for severe allergies or allergy-induced asthma.
cells and tissues—resulting in damage and discomfort.
TREATMENT PROFILE: ASTHMA
CELL SIGNALING PRIMER
Inhaled small molecule drugs have been the mainstay
Chronic inflammation is often caused by disorders in
for asthma sufferers for quite some time. Beta-
cell signaling, the system of communication governing
adrenergic receptor activators relax the smooth muscle
basic cellular activities and coordinating cellular
tissue in the lung, widening the airway and leading to
actions. Cell signaling is typically initiated by a signaling
easier breathing. Corticosteroids act as a general anti-
molecule binding to a receptor protein on the surface
inflammatory agent by decreasing the activation of
of a cell. This triggers a cascade of events inside the cell,
eosinophils, a type of white blood cell playing a major
ultimately resulting in a specific cellular response such
role in driving asthma-associated inflammation.
as cell growth and division, tissue maintenance and
repair, or immune cell activation. Newly approved Cinqair specifically targets eosinophilic
inflammation and is used on top of standard
The immune system makes a type of signaling molecule
medications to tame severe asthma. Cinqair is a
called a cytokine that modulates its activation response.
monoclonal antibody (mAb) that binds interleukin 5, the

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cytokine mentioned above that activates eosinophils. TREATMENT PROFILE: PSORIASIS
When interleukin 5 is bound by Cinqair, it is no longer
The flaky patches seen in psoriasis are skin cells simply
available to activate eosinophils, thereby reducing their
developing too fast. Even though psoriasis is seen on
overall numbers in the body and lessening the rate of
the skin, it is thought to be triggered by overactive
serious asthma attacks.
cell signaling spurring the immune system to cause
There is only one other mAb currently on the market, systemic inflammation.
Genentech’s (South San Francisco, CA) Xolair for
The newly approved mAb Taltz works by a mechanism
treating allergy-induced asthma. Xolair works by
similar to Cinqair. Taltz binds and inhibits interleukin 17,
inhibiting immunoglobulin E, the protein discussed
which activates the inflammatory pathways thought to
above that promotes allergic reactions.
drive psoriasis. With the cytokines blocked, the internal
There are several companies in late stage mAbs clinical inflammation is lessened, and the skin cells regenerate
development targeting the interleukins (IL) involved in at a slower rate.
eosinophil activation and general airway inflammation.
There are already a number of approved mAbs for
the treatment of psoriasis, all inhibitors of different
cytokines. Janssen Pharmaceuticals’ (Raritan, NJ)
Remade and AbbVie’s (North Chicago, IL) Humira both
work by impeding the inflammatory cytokine TNF-
alpha. Novartis’ (Basel, Switzerland) Cosentyx hinders
interleukin 17, while Janssen’s Centecor targets both
interleukin 12 and interleukin 23.

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UNMASKING MULTIPLE SCLEROSIS Life Science Training from Industry Experts

MANY CHANNELS AVAILABLE TO older, one theory points to these rogue WBCs being
IMPEDE MS PROGRESSION activated by a viral infection.

Continuing our series on central nervous system • Second, a breach in the blood-brain barrier (BBB),
(CNS) disorders—previously covering Alzheimer’s and the protective network of blood vessels preventing
Huntington’s—we pivot to unmask Multiple Sclerosis potentially harmful substances from entering the
this week. Famous faces suffering from Multiple brain. Exactly what elicits this compromise is not
Sclerosis (MS) include former talk show host Montel fully understood, but generalized inflammation—
Williams and Sopranos star Jamie-Lynn Sigler. MS possibly triggered by a viral infection—is thought
typically occurs in susceptible individuals between to play a role, in part because certain anti-
the ages of 20 and 50, and there are an estimated 2.3 inflammatory medications (see below) appear to
million cases worldwide according to the MS Society. lessen the permeability

Both a neurological disorder and autoimmune Other theories:

disease, MS originates as an immune-derived attack • Hormones may be involved—MS is more common
on the myelin sheath surrounding the axon portion in women than men by a three to one ratio.
of the neuron. With this insulating layer damaged, • Geography may play a factor—there is a higher
the electrical signal relied upon by neurons for incidence of cases the farther north one travels
communication loses the ability to push data forward. from the equator. Since sunlight is required
Initial symptoms vary due to the particular part of the for the sufficient production of vitamin D, it’s
nervous system affected. Neurons all along the front speculated the fat-soluble vitamin may protect
lines of communication are under threat, so symptoms predisposed individuals.
can include motor, sensory, or visual problems.
The ultimate expression of the disease is likely
About 85% of MS cases are relapsing-remitting (RRMS), the result of a combination of genetic and
meaning periods of recovery followed by remission environmental factors.
are seen between clearly defined periods of new or
increasing neurologic symptoms. Those afflicted with INJECTABLES BLUNT THE
RRMS usually transition into secondary progressive MS, IMMUNE RESPONSE
experiencing an accumulation of disability over time.
Although there is no cure for MS, there are a number of
Primary progressive MS, in contrast, exhibits a worsening
drugs on the market that can slow its progression. Most
of neurologic function from the onset of symptoms.
work by blunting the immune system’s attack on the
In both cases, significant loss in mobility continue as
CNS. MS sufferers switch between different drugs as
neurons lose their ability to send data.
the course of their disease worsens.
UNPLUGGING THE ROOTS One of the most common first-line treatments for
RRMS is recombinant interferon-beta: Avonex and
The identification of susceptibility genes, or genes
Plegridy by Biogen (Cambridge, MA), Rebif by EMD/
that increase the odds of developing a disease, have
Serono (Darmstadt, Germany) and Pfizer (New York,
contributed to the framework for understanding MS.
NY), Extavia by Novartis (Basel Switzerland), and
The direct cause, however, is largely unknown.
Betaseron by Bayer (Leverkusen, Germany). Interferon-
What we do know: beta is a signaling molecule that reduces the overall
• First, white blood cells (WBCs) must mistakenly levels of inflammatory agents in the brain. Patients on
label the myelin sheath as a “foreign invader.” interferon-beta therapy experience an increase in the
Since MS typically doesn’t develop until age 20 or integrity of their BBB, resulting in fewer myelin sheath
attacking WBCs entering the brain. Recent studies

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suggest interferon-beta may increase the production of IN THE CLINIC
nerve growth factor, improving neuronal survival.
MS drug discovery and development is very active. Here
Copaxone, marketed by Sandoz (Holzkirchen, Upper are just a few of the highlights:
Bavaria, Germany), consists of four amino acids—
• D
 aclizumab: Monoclonal antibody that binds to a
glutamate, lysine, alanine, tyrosine—that constitute a
receptor on the surface of T-cells, preventing them
large percentage of the myelin protein, which compose
from becoming fully activated. Regulatory filings
the myelin sheath. This injectable is thought to act as a
completed. Developed by Biogen and AbbVie
decoy of sorts to the immune system—white blood cells
(North Chicago, IL).
that would normally attack the myelin sheath attack
Copaxone instead. • O
 crelizumab: Monoclonal antibody targeting B-cells
for destruction. Previously, B-cells were not thought
Two monoclonal antibodies are available to treat MS.
to play a role in MS pathology, but the apparent
Sanofi’s (Paris, France) Lemtrada binds to a protein on
clinical efficacy of Ocrelizumab suggests they might.
the surface of T-cells, one type of WBC thought to be a
Currently in Phase III clinical testing. Developed by
key player in attacking the myelin sheath. A Lemtrada-
Biogen and Roche (Basel, Switzerland).
bound T-cell is targeted for destruction by other WBCs,
thereby reducing the number of T-cells available to • A
 nti-LINGO-1: LINGO is a protein that appears to
destroy the myelin sheath. Biogen’s (Cambridge, MA) inhibit myelination when it binds to its receptor on
Tysabri helps to prevent T-cells from entering the brain nerve cells. Anti-LINGO is a monoclonal antibody
by binding to proteins called VCAM1 on the surface attempting to stop this inhibition. Data from animal
of the blood vessels making up the BBB. In order for studies suggest it may even trigger re-myelination.
myelin-attacking T-cells to cross the BBB, they must If proven to work in humans, it will be the first MS
attach to these VCAM1 proteins and “pull” themselves drug to directly fix the myelin sheath. Currently in
through. With Tysabri in place, the T-cells cannot attach Phase II clinical testing. Developed by Biogen.
to the VCAM1 proteins, and are denied entry into • M
 N-166: Oral medication that suppresses the
the brain. production of pro-inflammatory signaling molecules
and increases the production of anti-inflammatory
SMALL MOLECULE FIGHTERS signaling molecules. In Phase II clinical testing.
Until 2010, injectable drugs (listed above) were the Developed by MediciNova (La Jolla, CA).
only option. With the approval of Novertis’ Gilenya,  ultiStem: Athersys (Cleveland, OH) is in preclinical
• M
MS medication in pill form arrived. Gilenya works by development of this “off-the-shelf” adult stem cell
binding to a receptor on the surface of T-cells that platform for the treatment of MS where tissue
prevents them from leaving the lymph nodes. Thus, matching isn’t required for treatment.
they are unable to enter the brain to weaken the
myelin sheath.
Two years later, a second oral MS drug, Sanofi’s
Aubagio, entered the market. Aubagio works by
inhibiting the growth of actively dividing cells—
including activated T-cells.
In 2013, much excitement surrounded the approval
of Biogen’s oral MS medicine, Tecfidera. Clinical trials
suggested it was at least as effective and had a better
safety profile than other MS drugs. While the exact
mechanism of action for Tecfidera is not known, it
has anti-inflammatory properties and activates the
expression of antioxidant proteins, which may explain
its neuroprotective effects.

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ON THE HUNT Life Science Training from Industry Experts

THE HURDLES OF HUNTINGTON’S

The nervous system is an incredibly complex piece of
human machinery, stretching to the far reaches of the
body while controlling and receiving the nuances of
life from a central command station. Just like any part
of the human body, the central nervous system (CNS)
is affected by various diseases that are sometimes
not entirely understood. After last week’s foray into
the Alzheimer’s, we turn to investigate Huntington’s
disease. Let’s catch up on the basics of the nervous
system and find out what biotech has in the pipeline for
this genetic neurodegenerative disease of the CNS.

NERVOUS SYSTEM PRIMER

Neurotransmitters are data delivery guys and work
The CNS—which consists of the brain and spinal cord—
in the synapse—the space between the neurons.
works in tandem with the peripheral nervous system.
When the dendritic branches of a neuron encounter a
The peripheral nervous system is the vast network
neurotransmitter, ion channels in the cell membrane
of nerves feeding into every tissue of the body, the
open, allowing positively charged sodium ions to enter
tentacles of data for the CNS. The signals received
the cell. The positive charge initiates an electrical signal
from nerve cells enable voluntary and involuntary
through the neuronal body and down the axon tail,
movement, and allow the brain to process and interpret
causing the release of other neurotransmitters into the
sensory information via the spinal cord.
next synaptic break. This process creates a cascade of
Specialized cells called neurons convert chemical neurotransmitters to react down the neuron chain, with
messages into electrical signals to convey data different neurons sending and receiving different types
throughout the nervous system. The branch-like of neurotransmitters.
extensions are called dendrites, which work to take
in chemical messages down through the cell body
(soma). The long, tail-like extension on the other end is
called an axon. They are enclosed in a fatty membrane
known as a myelin sheath, which serves to insulate the
axons and facilitate the passage of the electrical signal
through the axon terminals.

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The billions of neurons within the CNS—communicating trinucleotide repeat, but fewer than 36 repeats means
via hundreds of different types of neurotransmitters— the disease is not present. Higher than 40 means the
regulate just about everything within the human body: disease does manifest; copy numbers between 36 and
from movement to hunger, temperature, emotion, 40 means the individual may or may not be affected.
and wakefulness. Generally speaking, the higher the number of repeats
beyond 40, the earlier disease onset occurs and the
HUNTINGTON’S DISEASE EXPLAINED more quickly HD progresses.
Like Alzheimer’s, Huntington’s disease (HD) is also a
neurodegenerative disorder. Neurons progressively lose IN THE PIPELINE
structure and function, get damaged and eventually Until very recently, the function of HTT and how its
die. Early stages may show subtle symptoms, such mutations might cause neuronal cell death were largely
as involuntary movements and mood disturbances. unknown. Recent research from the University of
As the disease progresses and more neurons die, the California San Diego suggests that HTT plays a role
symptoms worsen. Sufferers of HD lose the ability to in activating gene expression in neurons through its
walk and speak, swallowing becomes more and more interaction with a protein known as PPARδ. PPARδ’s
difficult. Life expectancy is about 20 years after the function is significantly diminished in HD neurons,
onset of initial symptoms. 90% of HD cases affect adults possibly as a result of interaction with the mutated HTT
between the ages of 30 and 50; juvenile onset occurs in protein. Researchers identified an experimental drug,
the remaining 10% of cases. KD3010, as one that actually increases PPARδ activity.
HD is monogenic, it is caused by a mutation in one KD3010 was tested in a HD mouse model and found to
gene dubbed the Huntingtin gene. The disease is also significantly reduce neurodegeneration. Even better
autosomal dominant, meaning if one parent has HD, news—this drug has already passed human safety trials
their children have a 50% chance of getting it. There is as a potential diabetes treatment, so initiating HD trials
currently no cure for HD and some at-risk individuals of KD3010 could happen quickly.
struggle with the choice to test for the gene. Unless Because HD is a single gene disorder, it is also a good
symptoms of juvenile-onset HD are present, testing candidate for gene therapy and genome-editing
for HD before the age of 18 is prohibited to ensure approaches. Sangamo Therapeutics (Richmond, CA) is
those tested are old enough to understand the full conducting preclinical research on genome-editing in
implications of the disease. HD. Also in the mix is Ionis Pharmaceuticals (Carlsbad,
CA) with their Phase II clinical studies of an antisense
TERM OF WEEK: drug to block the production of the mutated HTT
TRINUCLEOTIDE REPEAT protein in HD patients.
The mutation that causes HD is known as a trinucleotide The hurdles of Huntington’s disease are high, however,
repeat, meaning the three nucleotides (CAG) are these latest technologies might just stand tall enough to
repeated multiple times in the middle of Huntingtin make a difference.
gene (HTT). Even normal versions of HTT have the CAG

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ATTACKING ALL ANGLES OF ALZHEIMER’S Life Science Training from Industry Experts

ALZHEIMER’S PIPELINE ROUNDUP Neuralstem’s lead neurogenesis candidate, NSI-189,

increased the hippocampal region of mouse brains by
Alzheimer’s disease ranks as one of the toughest nuts to as much as 20%. Neuralstem recently completed Phase
crack within drug discovery and development. Current I trials for major depressive order for NSI-189, with an
treatments merely manage symptoms, so finding a aim to branch out to Alzheimer’s disease in the future.
better solution becomes more and more urgent as the
aging population grows. YUMANITY’S YEAST
The pathology most commonly associated with Rather than directly targeting Aβ plaques, Yumanity
Alzheimer’s disease (AD) is the buildup of amyloid- Therapeutics (Cambridge, MA) is working to identify
beta (Aβ) plaques in the brain. Recent research from their roots. By engineering yeast cells to produce
Stanford University suggests the plaques bind to the Aβ protein, Yumanity monitors the detrimental
a receptor on nerve cells, disrupting their function. downstream effects of over-expression—like the
However, there is no absolute consensus that these disruption in the folding of other essential cellular
clumps of protein are the origins of AD or a symptom of proteins. Compounds that show promise in the yeast
the underlying cause. cells are then tested in AD patient-derived cells to
Most drug targets have focused on “mopping up” or screen for potential drugs. Yumanity is currently
inhibiting the production of Aβ plaques. Failures in preparing to begin clinical trials with its lead compound.
early clinical trials dominate the treatment landscape,
ANNEXON’S ACCUMULATIONS
with a few potentials in the pipeline—like Biogen’s
(Cambridge, MA) aducanumab in Phase III. In the race South San Francisco-based Annexon is bypassing Aβ-
to find a cure, every possibility offers a glimmer of plaques altogether and going after neuroinflammation.
hope, so let’s shine a light on the developmental drugs This pathway grew out of research conducted at
stepping away from Aβ plaques. Stanford, suggesting that a protein known as c1q is
present at higher levels in the brains of AD sufferers.
NEURONASCENT & C1q accumulates at neuronal synapses, the key points
NEURALSTEM’S NEURONS of communication between brain cells. C1q also acts
A key clinical feature of AD patients is the loss of as a flag for other immune cells like macrophages—
neurons. What if there was a therapy that could jump these “big eaters” chomp up cellular debris. The
start the development of new neurons? Neuronascent correlation of c1q could account for the observed
(Clarksville, MD) and Neuralstem (Germantown, MD) reduction in synapse numbers and the accompanying
are leading the charge in developing small molecule loss of cognitive function seen in AD. Annexon’s lead
activators of neurogenesis. By screening large chemical candidate ANX005, now in preclinical development, is a
libraries, they have identified various compounds monoclonal antibody which “mops up” excess c1q.
that show promise in activating neurogenesis from
PROTEOSTASIS’ PROTEASOMES
adult neural stem cells, both in tissue culture and in
mouse models. In partnership with Biogen, Cambridge-based
Proteostasis Therapeutics is targeting AD-associated
In a mouse model of Alzheimer’s, Neuronascent’s
protein aggregates by activating a cellular component
lead compound NNI-362 promoted the growth of new
known as the proteasome. Proteasomes get rid of
hippocampal neurons that not only migrated to the correct
damaged proteins and dysfunctional protein aggregates
functional location but also differentiated and survived
by dismantling the peptide bonds holding them
long enough to reverse previously observed cognitive
together. USP14 is one of the proteins that inhibits the
declines. The hippocampus is thought to play a role in
proteasome, so naturally Proteostasis is working on the
memory formation and spatial navigation and is one
preclinical development of a USP14 inhibitor to allow
of the first regions of the brain to show damage in AD.
proteasomes to be fully activated in AD patients.
Neuronascent is preparing for Phase I trials of NNI-362.
Attacking Alzheimer’s from all angles is the surefire way
to get closer to better treatments and a real cure.

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THE EMERGING PROTEOMICS MARKET Life Science Training from Industry Experts

is organized into databases and applied to medical and

biological questions.

GENOME VS. PROTEOME

The genome is fixed for life—it’s the same at birth
as it is at age 99. And that genome is the same in
every cell of your body—this is why sequencing the
genome of a white blood cell, for example, can yield
genetic information about a patient’s liver. Of course,
environmental exposures to certain chemicals or
radiation may cause changes in the DNA of some cells,
but for the most part, the genome is fixed.
The proteome varies from cell type to cell type. Even
though a liver cell and a white blood cell have the same
DNA sequence, each cell has unique characteristics
and functions. Each cell type turns on and off distinct
sets of genes and it is this feature that confers different
cell characteristics.

PICKING APART THE PROTEIN The proteome is fluid over the course of a lifetime
with constant change in protein production through
Genomics continues to be all the rage in biotech
the years—and in some cases throughout a day or
circles—with special kudos to Illumina’s (San Diego,
hour. The amount of a specific protein often changes
CA) recent ability to obtain an entire human genome
significantly between healthy and diseased states.
sequence in 24-hours for a mere thousand dollars. Our
Some illnesses are the result of an increase in the
overall understanding of human DNA combined with
expression of proteins, such as the over-expression of
our ability to determine individual genomes leads to
HER2 in HER2-positive breast cancer. Other illnesses are
better disease insight, more powerful diagnostics, and a
the result of a decrease in the expression of proteins,
higher output of efficacious therapeutics.
such as the decreased expression of insulin in type
Equally important, though not as well-understood, 1 diabetes.
is the field of proteomics. While genomics testing
Importantly, these changes in the proteome can exist
lays out the odds of developing a particular disease,
even when there are no differences in a diseased
proteomics testing reveals the presence of disease and
person’s genome in comparison to a healthy individual.
its stage of development. In this issue, we’ll pore over
Developing a better comprehension of the proteome
the proteome and prime our readers for this emerging
in both diseased and healthy states is critical for truly
biotech market.
understanding disease and developing new diagnostics.

TERM OF THE WEEK:

PROTEOME & PROTEOMICS STUDYING THE PROTEOME:
MASS SPECTROMETRY
The proteome is the full complement of proteins
Determining the identity and relative concentrations
produced—or, expressed—by the genes of a particular
of different proteins in biological samples can be
cell, tissue, or organism. Proteomics is the analysis
technically challenging. One tried and true approach is
of the structure, function, and interactions of those
mass spectrometry. This complex technology can be
proteins. The big data generated by a proteomics study
summarized as a series of steps:

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1) Protein samples are treated so their complex three- colorectal cancer. Integrated Diagnostics’ (Seattle,
dimensional structures are unfolded, making them WA) blood test determines the probability that a
easier to analyze. lung nodule is benign by measuring levels of multiple
2) Unfolded proteins are “digested” with enzymes that circulating proteins.
cut them up into smaller fragments called peptides.
A TWIST ON PROTEIN MICROARRAYS
3) The peptide fragments are “ionized,” or given an
Protein microarrays are another way to determine
electrical charge. The process of ionization further
the identity of proteins. Traditionally, antibodies are
fragments the peptides.
attached to a glass chip. Each antibody has an affinity
4) Positively charged peptide fragments move through a for a specific protein. When samples are washed
magnetic field towards a negatively charged panel. over the chip, proteins will attach to their respective
5) Peptide fragments with different masses move at antibodies. Once “captured,” the proteins are decoded.
different rates, creating a graph or “mass spectrum” SomaLogic (Boulder, CO) is bringing a new twist to
which is compared to the baseline spectrum in the microarray game. Instead of using antibodies as
a database. the “capture” agent, they use DNA sequences called
6) The comparison identifies the proteins present in the aptamers, which are selected in the lab for their affinity
initial sample. to specific proteins. These aptamer sequences are
A number of companies have developed diagnostic attached to microscopic beads and probed with the
tests based on variations of mass spectrometry: protein sample. The aptamer sequences that capture
Biodesix’s (Boulder, CO) VeriStrat test measures the proteins can be easily quantified using techniques such
levels of different proteins in a blood sample of lung as qPCR. SomaLogic markets an inflammation panel
cancer patients to identify those with more aggressive which determines the levels of sixteen inflammation-
cancers. The VeriStrat test also reveals which patients related proteins.
will respond best to Genentech’s (South San Francisco, As the field of proteomics evolves, we can expect to
CA) Tarceva. Applied Proteomics’ (San Diego, CA) see a more comprehensive understanding of disease
SimpliPro analyzes eleven proteins correlated with translated into superior diagnostics and therapeutics.

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BIOTECH BATTLES ZIKA Life Science Training from Industry Experts

LATEST MOSQUITO-BORNE The biggest concern is for pregnant women—Zika has

been linked to microencephaly, in which babies are
DISEASE OUTBREAK
born with brains significantly smaller than normal.
The buzz around the Zika virus isn’t going to die down
Microencephaly results in developmental delays,
anytime soon. The global health threat is dominating
intellectual disability, seizures, difficulty swallowing,
the news while the Brazilian outbreak threatens to
hearing loss, and vision problems (CDC). The
spread to the rest of the Americas.
mechanism by which Zika causes microencephaly is not
As the ramifications of the virus emerge every passing known, although it has been confirmed that the virus
day, we here at Biotech Primer WEEKLY wonder: what is able to pass through the placenta. This is known as
is biotech doing to fight Zika? In this issue, we’ll vertical transmission, or the passing of a virus from
examine the virus and learn how companies plan to mother to fetus.
exterminate Zika.
GMO MOSQUITOS TO THE RESCUE?
TERM OF THE WEEK: VECTOR Intrexon (Germantown, Maryland) is taking a high-
Epidemiologists call an agent that carries and transmits tech spin on mosquito control by creating genetically
a pathogen into another living organism a vector. The engineered Aedes aegypti mosquitos in the lab. The
Aedes aegypti mosquito serves as the vector for Zika mutants contain a gene that codes for a protein which
virus, as well as dengue virus and yellow fever virus. prevents the expression of other genes critical for
survival. When kept in the lab, the male mosquitos are
THE ZEAL OF ZIKA
fed an “antidote” which suppresses the expression of
Sporadic cases of Zika have been detected in humans
the lethal protein. When released into the wild, the
since the 1950s in equatorial regions of Africa and Asia.
male mosquitos will survive long enough to mate and
In 2007, the first major outbreak occurred in Micronesia,
pass this gene on, however, their offspring do not
with 49 confirmed cases reported. The current outbreak
survive. The GMO mosquitos are currently making their
began in Brazil in April of 2015. The virus has now
way across Brazil, Panama and the Cayman Islands to
spread to other countries in South and Central America,
complete efficacy trials. According to Intrexon, these
the Caribbean, and Mexico.
studies show more than 90% reduction of the Aedes
The main route of Zika virus transmission is through aegypti mosquito population.
the mosquito Aedes aegypti. When the mosquito bites
an infected person, blood containing millions of tiny AN OUNCE OF PREVENTION
virus particles is taken up, the virus multiplies, and Since there is no vaccine available for Zika, several
penetrates into the mosquito’s salivary glands. When an companies have jumped at the challenge to prevent
infected mosquito bites its next victim, Zika is injected future outbreaks. Inovio Pharmaceuticals (Plymouth
and the person becomes infected. A less common route Meeting, PA), known for its expertise in DNA vaccine
is the sexual transmission from an infected human. development, has recently begun a new Zika-focused
To date, the CDC has three confirmed and fourteen vaccine program.
suspected sexually-transmitted Zika cases in the U.S. Hyderabad, India-based Bharat Pharmaceuticals has
For most, the virus causes mild symptoms such as fever, been working on a Zika vaccine since November 2014—
skin rash, and joint pain that go away in a few days with before the current outbreak started. With both a DNA
rest. However, public health officials report there may vaccine and a more traditional inactivated virus vaccine
be an association between Zika and a slight increase in ready for testing, Bharat is currently seeking approval
cases of the rare neurological disorder Guillain-Barre from the Indian government to begin preclinical trials
syndrome, a rapid-onset of muscle weakness caused by in animals.
the immune system damaging the peripheral nervous The race to beat the spread of Zika is on. By attacking
system. Guillain-Barre syndrome is known to be the outbreak from all angles, we have a better chance
triggered by certain viral infections. containing—and maybe even exterminating—a
potential world health crisis.

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STOMACH STAPLING PAVES A PATHWAY Life Science Training from Industry Experts

IDENTIFYING GUT PROTEINS TO It appeared that rerouting the digestive tract altered
TACKLE METABOLIC DISORDERS some basic physiology. Various hormones are
released in response to digestion, signaling satiety
Historians of science love to tell about “eureka
and regulating the pace at which the stomach empties.
moments”—simple observations leading to major
Could one of these hormones be altered just by
insight. Greek mathematician Archimedes allegedly
rerouting the stomach? This question formed the basis
exclaimed the original “Eureka!”—meaning “I have found
of NGM Biopharmaceuticals, a company founded on
it!”—upon witnessing the volume of water displaced
identifying potential therapeutic proteins produced in
by his body in the bathtub equaled the volume of the
the gastrointestinal (GI) tract.
body part he had submerged. Medicine’s most famous
eureka moment is probably the discovery of penicillin in REPLICATING A STOMACH HORMONE
1928 when microbiologist Alexander Fleming observed
One of the first GI tract proteins with potential
the absence of bacteria developing on his growth plates
metabolic benefits identified was fibroblast growth
in areas where the mold Penicillium rubens happened
factor 19 (FGF19). FGF19 is a hormone whose primary
to be sprouting.
function is to regulate bile synthesis, but FGF19 also
The roots of NGM Biopharmaceuticals (South San impacts glucose and lipid metabolism by interacting
Francisco, CA) are based on a eureka moment as with receptors on the surface of fat cells. In fact,
well—the simple observation that many recipients of patients with metabolic syndrome, non-alcoholic fatty
gastric bypass surgery begin to see almost immediate liver disease, or insulin resistance all have reduced
improvement in blood glucose levels before the levels of FGF19—which is restored to normal levels after
occurrence of weight loss, and in some cases even gastric bypass surgery and before weight loss. This
without significant dietary changes. In this issue, suggests lowered levels of FGF19 may be contributing
we’ll find out how the science of gastric bypass to some of the observed metabolic problems of
surgery is being used to discover new drugs in the obese patients, and increasing those levels may
metabolic space. confer benefit.

BYPASSING NORMAL NGM Bio’s drug candidate NGM282 is an engineered

DIGESTIVE ROUTES version of FGF19. NGM282’s gene sequence is
modified, so it does not promote liver cell growth
Gastric bypass surgery is pretty much “stomach
but still interacts with the signaling pathways
stapling” and then a change-up in the route of
regulating bile synthesis and normalizing glucose and
digestion. A portion of an obese patient’s stomach is
lipid metabolism.
made physically smaller, and therefore only capable of
holding about a cup or so of food at a time. Then the The exact mechanism by which FGF19/NGM282
new, smaller stomach is disconnected from the upper works is not fully understood. It is known that both
part of the small intestine and reconnected lower down, proteins interact with receptors on the surface of liver
thereby reducing absorption of food through the small and fat cells, and likely influence signal transduction
intestine. Less food and less absorption of food lead to pathways. Recent research also suggests that FGF19
weight loss. (and by extension, NGM282) also interact with neuronal
pathways that regulate metabolism.
The initial understanding of surgery results implied
positive outcomes came from weight loss (averaging
60% of excess weight). However, the significant
improvement in blood glucose levels seen in many
patients immediately following the surgery—before
weight loss occurs—remained unexplained.

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 ANOTHER GASTRIC PATHWAY?
FGF19 is not the only hormone released in the gut
(stomach, pancreas, and small intestine) that plays a
critical role in glucose and fat metabolism. More than
twenty exist to control appetite, glucose absorption,
and insulin release. These complex interactions help to
explain why understanding and managing metabolic
disorders continue to be a challenging medical problem.
One of the best characterized gut hormones is
glucagon-like peptide 1 (GLP-1). Secreted by cells of
the large intestine, GLP-1 increases insulin secretion
and insulin sensitivity, inhibits stomach emptying, and
decreases glucagon secretion. Since insulin release
causes muscle, fat, and liver cells to absorb blood
CHALLENGING CHOLESTATIC LIVER
glucose, and glucagon stimulates the liver to release
DISEASE, NASH & DIABETES
stored glucose, increasing insulin while decreasing
NGM282 is currently in Phase II clinical trials for glucagon will reduce blood glucose levels.
cholestatic liver disease, an inflammatory liver disease
There are several Type 2 diabetes drug on the market
caused by the build-up of bile in the liver. NGM282
acting as GLP-1 analogs, or peptides (amino acids
appears to modulate the production of bile acid,
sequences composed of 50 or fewer amino acids).
resulting in reduced liver inflammation. The drug is
These drugs mimic the effect of GLP-1 and lower
also in Phase II for Type 2 diabetes due to its ability to
blood glucose levels—examples include Amylin
lower blood glucose levels. Look for Phase II trials for
Pharmaceutical’s (San Diego, CA) Byetta and Lilly’s
nonalcoholic steatohepatitis (NASH) in the near future.
(Indianapolis, IN) Trulicity.
The rise in metabolic disorders has ballooned, so let the
eureka moments continue!

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FINDING FOOTPRINTS Life Science Training from Industry Experts

NEW PLAYER IN CANCER

BIOMARKER DETECTION
Catching a tumor’s first footprints is the holy grail of
cancer research. The ability to detect and treat cancer
early would put the collective mind of humanity at
ease. Our best bet yet is the liquid biopsy—a minimally
invasive test to detect and monitor cancer, which
happens to be all the rage in cancer diagnostics.
Liquid biopsies uncover the beginnings of a tumor
from tell-tale clues cancer cells leave behind in blood
or urine. Examples include DNA released from dying
cancer cells, exosomes secreted from cancer cells, and (Photo credit: Chris Woodcock, via Wikimedia Commons)

even rogue cancer cells themselves broken off from

a tumor. EPIGENETICS EXPLAINED
A new player on the block, Belgium-based VolitionRx, Changes to DNA that do not alter the actual gene
is developing liquid biopsies for colorectal and other sequence, but rather are chemical modifications to the
cancers based on a novel biomarker: nucleosome DNA itself are the basis of epigenetics. These changes
structure. In this WEEKLY, we’ll tune in to the science typically affect gene expression, or how the gene is read
behind nucleosomes and examine its role in by the cells. Epigenetic modification can occur either
cancer diagnostics. directly to the nucleotide bases themselves (A, C, G,
or T) or to the histones, which are small proteins that
TERM OF THE WEEK: NUCLEOSOME package and order DNA.
If we unraveled the DNA out of a single human cell, One of the most common types of epigenetic
we’d have a six-foot long double helix. DNA undergoes modification is methylation—the addition of a methyl
extensive “packaging” to fit inside of a cell—it is (CH3) group to cytosine (C) nucleotides. The result:
wrapped around proteins called histones, in much the methylation reduces or even blocks gene expression.
same way thread winds around a spool.
A second type of modification is called acetylation—the
The basic unit of DNA packaging is called a nucleosome, addition of an acetyl group (CH3 CO) to the histones.
which consists of a segment of DNA wound around Acetylation loosens the association of the DNA with
eight histone protein cores. The DNA in our cells is a the histones, making the DNA more accessible to the
series of nucleosomes connected by linker DNA, or enzymes used in gene expression, ultimately increasing
DNA not associated with histone proteins. The phrase protein production.
“beads on a string” is often used to describe the final
Deacetylation—the removal of an acetyl group—
configuration. A picture of nucleosome structure
increases the association or “grip” of the DNA around
captured by an electron microscope is shown below:
the histone proteins. Deacetylation makes the DNA less
accessible to enzymes used in gene expression, thereby
decreasing the production of proteins.
Histone proteins may also undergo a variety of other
chemical modifications, including the addition of a
phosphate group (phosphorylation), or ubiquitination.
The exact combination of epigenetic modifications will
determine the impact on gene expression.

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UNREELING NUCLEOSOMICS The nucleosomes are then probed with antibodies that
recognize cancer-associated epigenetic modifications.
Since epigenetic differences impact gene expression—
The extremely high specificity of antibodies
how much or how little of a particular protein is made—
allows for such precision—researchers are able to
it’s not surprising that various epigenetic changes are
develop antibodies which differentiate between
associated with different cancers. These epigenetic
the same histone proteins with different epigenetic
changes can be detected by examining nucleosomes.
modifications. The antibodies used as protein-specific
When cells (including cancer cells) die, enzymes in
probes will typically have a fluorescent dye so the
the cell’s nucleus cut the “linker DNA” connecting one
presence of the disease-associated protein can be
“bead” (nucleosome) on a string to the next. These
detected by the change in color. VolitionRx’s test is able
nucleosomes are then released into the patient’s blood.
to differentiate between their testable cancers and
Diagnostics based on collecting nucleosomes from other nucleosome-elevated conditions.
a blood sample and looking for specific structural
changes—epigenetic modifications —associated with
cancer are in development by VolitionRx (Namur,
Belgium). The test requires a very small amount of
patient blood for analysis: approximately 10 microliters
(one one-thousandth of a liter) as opposed to 5,000
microliters for other types of liquid biopsies. The
company recently reported data demonstrating over
90% accuracy in colorectal, lung, and pancreatic cancer
screenings, and is conducting additional studies in
colorectal, lung, and prostate cancer.

SEIZING THE FIRST SIGN

How do the tests work? The first step is to isolate
nucleosomes out of the blood by using an antibody that Antibodies find and attach to cancer-associated nucleosomes.
recognizes and binds to a core histone protein common
to all nucleosomes. The pairing “pulls” the nucleosomes The new focus on liquid biopsies to uncover the hidden
out of the blood. biomarkers of cancer might be the big win in stopping
cancer early. With a slew of players in the race, it’s only
a matter of time before we find the holy grail.

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DRUGGING THE UNDRUGGABLE Life Science Training from Industry Experts

ANTAGONISTS FIGHT A GOOD FIGHT

Small molecule inhibitors, also known as antagonists in
the industry, fight a good fight. In fact, many drugs on
the market today work by inhibiting overactive, disease-
associated proteins. Novartis’ (Basel, Switzerland) top
selling leukemia drug Gleevec, for example, is a small
molecule inhibitor of a protein called Bcr-Abl, whose
overactivity promotes excessive cell division.
The downside? It is hard to develop effective small
molecule inhibitors. The best inhibitors are designed
by knowing the shape of the target—information which
is often difficult to get. Even if researchers do know
the shape of the target, it may not be one for which
an effective inhibitor can be designed. This is because
the part of the protein that is overactive, for example,
may be a smooth or flat surface and inhibitors need
something to “hook onto”. And for those proteins for
which scientists are able to develop strong inhibitors,
resistance may emerge, rendering the drug ineffective.
MEDDLING IN THE MIDDLE
What if, instead of inhibiting disease-associated
proteins, we could simply dump ‘em? It turns out, Arvinas (New Haven, CT) is developing a platform
cells already have a system for getting rid of defective to target disease-causing proteins based on the
proteins. So, let’s unwrap a new class of drugs taking ubiquitin and proteasome interaction. Dubbed PROTAC
advantage of these tiny cellular garbage disposals and (Proteolysis-Targeting Chimera), the technology consists
find out how we might just drug the undruggable. of “bifunctional small molecules”—small molecules
that have been designed to simultaneously bind to two
LIKE A GARBAGE DISPOSAL different things.

All cells contain specialized compartments called In the case of PROTAC, one end binds to the target
proteasomes whose job is to degrade unneeded disease-causing protein and the other end to the
or damaged proteins. Like a garbage disposal, E3 ligase. This interaction triggers the transfer of
proteasomes are hollow on the inside in order to an ubiquitin protein to the target, marking it for
enclose and break down the peptide bonds that hold degradation. The whole complex is then degraded and
the proteins together. dumped by the proteasome. Since the PROTAC doesn’t
necessarily have to recognize a very specific part of the
Proteins are targeted for degradation via the action of
target protein, it is possible to target a wider range of
an enzyme called E3 ligase. E3 ligase first instructs the
proteins compared to existing technologies.
protein ubiquitin to attach to the unneeded protein.
Then, ubiquitin guides the protein into the proteasome Arvinas has released preclinical results on their
chamber, where it is broken down for disposal. platform’s ability to target the protein BRD4, which
This process enables the cell to regulate protein plays a role in cell division. Mutated versions of BRD4
concentration and dump any misfolded proteins. are associated with various cancers. Data suggests
PROTAC has successfully lowered levels of the protein
in lymphoma, multiple myeloma, and prostate
cancer cells.

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TERM OF THE WEEK: DRUGGABILITY RNA TO THE RESCUE?
Druggability refers to whether or not it is possible to RNA-based drugs are another approach to putting a
develop a drug that will bind to a particular target, and dent in the undruggable proteins of today. Recall from
alter the function of that target in a disease-modifying high school biology that RNA translates DNA code into
way. The structure of the target is a key determinant a language ribosomes can understand in order to make
of druggability. It’s been estimated that only about 2% proteins required by the cell. Messenger RNA (mRNA) is
of human proteins interact with currently approved the intermediary between genes and proteins: a gene
drugs, and only about 10% to 15% of human proteins is transcribed into mRNA, and that mRNA is translated
are druggable using today’s technology—and these into chains of amino acids—the building blocks of
druggable proteins may or may not actually be disease- proteins. Because these drugs target the mRNA
associated (source). These numbers underscore transcript of a protein rather than the protein itself,
the importance of developing new technologies for they do not require a protein structure that is accessible
drug discovery. to drugs. Read more about RNA-based platforms like
The disease-associated proteins inhibited by RNAi, antisense and more in our previous issue.
antagonists on the market are druggable proteins. But,
because antagonists are hard to make, and resistance
may develop over time, the need for additional avenues
is clear. Platforms such as PROTAC—and even RNA-
based technology—circumvent the need to inhibit a
protein by halting the production of the protein in the
first place.

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INTERRUPTING HIV Life Science Training from Industry Experts

THERAPIES IN HIV PIPELINE

Medicine has made gigantic strides in understanding
and treating the human immunodeficiency virus (HIV)
since its emergence in the early 1980s. Thanks to
robust drug discovery efforts, today HIV is managed
as a chronic disease instead of the death sentence of
years past.
The secret to HIV’s infectious success lies in its high
mutation rate. Developing drug resistance over time
is the name of the game as viral proteins change just
enough to escape detection, keeping researchers
on their toes. In this issue, we’ll examine the drug
industry’s persistence to stay ahead of HIV.

The mechanisms of action detailed above show the

HIV PRIMER various ways approved drugs target HIV.
HIV causes acquired immunodeficiency syndrome
(AIDS) because it infects and disables T-cells, a critical IN DEVELOPMENT
type of white blood cell. This incredibly destructive
Bristol-Myers Squibb (New York, NY) has begun Phase
power is carried out by just a handful of proteins
III clinical testing of BMS-663068 which targets GP120,
and a lipid envelope encapsulating the RNA-based
the portion of the viral surface protein that enables
genetic material.
HIV to bind to and enter T-cells. A similar drug target—
but only by proximity—is Roche’s (Basel, Switzerland)
injectable Fuzeon, which targets the GP41 portion of
the same viral surface protein. Even though both of the
drugs are targeting nearby areas, the part of GP120
targeted by BMS-663068 mutates at a much lower rate
than GP41, adding hope that resistant strains of the
virus will be less likely to emerge.
All of the HIV drugs currently on the market are small
molecule drugs, except for Fuzeon, which is a peptide,
or a short chain of amino acids (the building blocks of
proteins). Investigators at Rockefeller University (New
York, NY) in collaboration with Celldex Therapeutics
(Hampton, NJ) have initiated clinical trials of “broadly
neutralizing” antibodies, or antibodies that recognize
HIV infiltrates T-cells in order to expand its grip on the immune system.
multiple strains of HIV, as both a therapeutic and a
First introduced in the late 1980s, HIV drugs have been preventative measure.
developed on scientists’ understanding of the viral life
cycle. By deciphering key steps in viral infection and
replication, researchers have been able to come up
with drugs to stop the virus. Most HIV patients take a
combination of drugs to control infection.

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THE VACCINE HOPE MAKING THE RIGHT CUT
The best way to prevent any infectious virus is The dance to stay ahead of HIV lingers on, but a
through vaccination. Researchers have been trying new possibility might just stop the music. Sangamo
to successfully develop an HIV vaccine for decades. Biosciences (Richmond, CA) is currently conducting
Unfortunately, the same high mutation rate that Phase II clinical testing using genome editing to
empowers the virus to become resistant to drugs also eradicate the virus from infected patients. The idea is
enables it to evade immune responses potentially simple: remove T-cells from infected patients and edit
activated by vaccination. their genomes so that their T-cells no longer contain
Dr. Robert Gallo—a scientist who identified HIV as the CCR5 receptor. These edited T-cells are infused
the cause of AIDS back in 1984—is pioneering a new back into the patient’s body, where they reproduce
approach to HIV vaccination at the University of and become the primary T-cell population. No CCR5
Maryland’s Institute of Human Virology. Traditional receptor, no HIV infection. No virus can survive outside
vaccines work by injecting patients with an inactivated of its host cell, so by making T-cells impossible to infect,
version of the virus, or with one protein from the viral the virus no longer has a host and will not survive.
surface, in hopes of training the immune system to Genius!
produce antibodies to recognize and target the virus.
When that target is constantly changing, training the
immune system is somewhat of a moot point.
Instead of targeting the virus in its resting state, the
University of Maryland vaccine targets the virus when it
is in a transition state: the precise moment at which the
virus binds to its CCR5 receptor on T-cells. During this
transition, normally hidden parts of the GP120 protein
are exposed. Because this part of the protein is critical
for viral entry, it has a very low rate of mutation since
most random mutations would make the virus less
effective in cell entry.
Since this part of the GP120 protein is normally hidden,
the immune system will not produce antibodies
against it if the intact virus or GP120 protein is used
as the vaccine. The trick is to stabilize the virus in the
Sangamo Therapeutics is using its genome-editing platform
transitional state by linking the GP120 protein to a to disrupt the CCR5 gene on patients’ T-cells.
portion of the CD4 receptor. This forces the virus to
take on the conformation in which the “hidden” parts The leap from HIV being an unmanageable disease to a
are exposed, enabling the immune system to produce manageable one took a few decades. With the advent
antibodies against this critical portion of the GP120 of new biotechnologies on the rise, it may be just a
protein and prevent infection. few more hops before we land on a true breakthrough
against HIV.
After years of testing in various animal models,
University researchers—in collaboration with Profectus
BioSciences (Baltimore, MD)—announced they were
starting Phase I human clinical testing on the novel
vaccine last October.

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DETECTING A SILENT KILLER Life Science Training from Industry Experts

GETTING TO THE HEART OF USING GENES TO BREAK THE SILENCE

SUDDEN CARDIAC ARREST Chicago-based 3PrimeDx is developing a molecular
Sudden Cardiac Arrest (SCA) doesn’t discriminate against diagnostic to identify those at risk for SCA based on
anyone. From healthy to unhealthy, young to old, the silent genetic factors. The test borrows from research
unexpected cessation happens in an instant. Affecting demonstrating that people who express a shortened
more than 325,000 people per year with a 90% fatality version of the protein SCN5A are at greater risk for SCA.
rate, it is one of the leading causes of death among Since expression levels of different SCN5A variants in
people over 40. white blood cells mirror expression levels in cardiac
Individuals determined to be at risk—those with previous tissue, the test can be done via a simple blood draw.
heart attack, coronary artery disease, and heart rhythm The variants in SCN5A expression arise due to a process
disorders—may be fitted with an a battery-powered called “alternative splicing”, wherein various parts of the
device called an implantable cardioverter defibrillator. RNA produced from the SCN5A gene are spliced together
It is placed under the skin and tracks heart rate. If an differently in at-risk individuals. This splicing pattern
abnormal heart rhythm is detected, the device delivers is not necessarily constant throughout life, but studies
an electric shock to restore a normal heartbeat. suggest that detection of aberrant splice variants in
About 20% of SCAs have no risk factors and strike white blood cells can be used to predict increased SCA
people in the prime of their lives. In most cases death risk within the next year.
is instant, with survival possible if CPR or electric shock
to the heart are delivered within minutes. In this issue, SCN5 EXPLAINED
we’ll take a closer look at SCA and highlight a molecular SCN5 is a sodium channel—a protein embedded in
diagnostic test in the works to identify people at risk for cardiac muscle cell membranes to let positively charged
this silent killer. sodium ions enter and exit. Proper control of ion flow
is critical to maintaining consistent heart rhythm, so it
EASILY CONFUSED: SCA makes sense that defects in this protein are correlated
VS. HEART ATTACK with SCA.
SCA is an electrical problem—the heart stops beating
due to an electrical malfunction. Sometimes, victims JUMPSTARTING THE HEART
of SCA have warning signs such as fatigue, fainting, The best treatment for SCA is immediate defibrillation,
blackouts, dizziness, chest pain, shortness of breath, or the delivery of a therapeutic dose of electrical current
weakness, palpitations or vomiting, typically within to the heart. This essentially “resets” the heart, allowing
24 hours of cardiac arrest. In others, there are no a normal rhythm to be reestablished. Automated
warning signs. external defibrillators (AED) are available in hospitals,
A heart attack is a plumbing problem— the heart is ambulances and some public places; however, time is of
still beating, but a blocked artery prevents blood from the essence in cases of SCA.
reaching a section of the heart. The longer the flow For those at known risk for SCA, implantable
of blood is cut off, the more severe the damage to the cardioverter defibrillators (ICDs) are a potentially
affected area. It is imperative to start treatment with lifesaving option. These devices have the ability to
anti-clotting medications or cardiac catheter as soon as detect and correct life-threatening cardiac arrhythmias,
possible to reduce the clot and free up the artery. including SCA. With 3PrimeDx’s diagnostic test aiming
to identify those at risk for SCA, ICD placement could be
the next step after diagnosis. 3PrimeDX’s test is currently
in development and the company is expected to release
more data in 2017.

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THE MEDICINE MACHINE Life Science Training from Industry Experts

MODERN TECHNOLOGY The platform involves taking the basic chemical building
& MOTHER NATURE blocks found in the original molecule and recombining
them in ways that maintain the original structure—but
For thousands of years, nature has been the best
rearranged enough to improve efficacy or remove toxic
medicine cabinet around. Natural products are drugs
components. The actual machine used in the process,
derived from nature, typically plants or microbes, and
called REVBLOCKS, relies on a type of reaction worthy
have been especially useful in fighting cancer and
of the 2010 Nobel Prize in chemistry—carbon coupling.
infectious disease.
Carbon coupling uses metal ions as catalysts, and can
Drugs from nature can be highly effective. Think be performed at room temperature and without other
Bristol-Myers Squibb’s (New York, NY) Taxol, the potentially toxic additives.
breast cancer drug derived from the sap of the Pacific
The bottom line? Figuring out how to synthesize Taxol
yew tree. Another is Artemisinin, which is procured
took over a decade. Now, chemical synthesis pathways
from the plant Artemisia annual and used to treat
that used to take years to design can be completed in
malaria. Even good ol’ aspirin was originally taken
days, or even hours.
from willow tree bark and later synthesized by Bayer
(Leverkusen, Germany). Today, more than 50% of drugs TAKING THE TERRIBLE OUT
on the market are natural products or their derivatives. OF AMPHO-TERRIBLE
Many natural products show great promise in the lab, Over the past several decades, our heightened capacity
but are not easily translatable into new medicines to care for immunocompromised and significantly ill
because they are too difficult to produce on a large patients has led to an increase in fungal infections.
scale, or they prove to be toxic to humans. In this issue, Resistance to existing antifungals has become the
we’ll examine how we are overcoming these barriers norm, leading to more reliance on amphotericin B, a
thanks to innovative companies combining modern highly potent but extremely toxic drug. Side effects
technology with mother nature. ranging from fever and chills to impaired kidney and
liver function—and even cardiac failure—precipitated
REBUILDING MOLECULES the nickname of “ampho-terrible” to the last
WITH DR. BURKE resort antifungal.
Drugs from nature have definitely hit a few roadblocks.
Dr. Burke’s research uncovered the source of this
Some are highly toxic—not only do they kill cancer
toxicity: amphotericin B is effective because it
or bacterial cells, but they also wipe out human cells.
leeches ergosterol. Ergosterol is the fungal version
Others are highly effective and potentially safe, but
of cholesterol and a critical component of fungal cell
prove to be very difficult to reverse-engineer. It is tough
membranes. It binds to and destroys fungal cells in the
to produce a synthetic pathway to make large quantities
human body by seeking out the ergosterol found in
of a drug in a lab without relying on scarce and often
the fungal cell membranes. Unfortunately, because of
difficult to collect natural products.
similarity between human cholesterol and ergosterol,
Enter Revolution Medicines (Redwood City, CA). it also manages to destroy cholesterol molecules—and
Founded by Dr. Martin Burke (no relation!), the therefore human cells—at the same time.
company’s new approach uses technology licensed
Using the REVBLOCKS platform, Revolution created a
from the University of Illinois. Revolution has
safer version of amphotericin B that solely recognizes
developed a machine capable of taking molecules
the fungal version of cholesterol, leaving human cells
showing therapeutic potential and rendering them both
intact. The company has seen promising preclinical
safe and effective.
results and is preparing for clinical trials.

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COCKTAIL FODDER: ANOTHER CELL’S behind Chematica, a software package that has
TRASH IS A PHARMA TREASURE been dubbed the “chemical internet” or “Google for
Chemistry.” Researchers can enter a molecule they want
Synthesis of the influenza drug Tamiflu relies on a
to synthesize into Chematica and the software sifts
chemical called shikimic acid. Shikimic acid is derived
through 200 years of organic chemistry know-how to
from star anise, the fruit of the plant Illicium verum,
recommend the best recipe. Currently, the software is
an evergreen tree found in Vietnam and China. During
used by a few research groups, with commercialization
the 2005 and 2009 swine flu outbreaks, star anise
plans in the works.
shortages created Tamiflu shortages, so Roche (Basel,
Switzerland) begun using bacterial fermentation to Also in the mix is Dial-A-Molecule, a British project
make shikimic acid. When a particular strain of E. led by University of Southampton chemist Richard
coli is overfed sugar, shikimic acid is produced as a Whitby with the goal of encouraging collaboration
waste product. among different research groups to develop automated
chemical synthesis machines. The boom of scientific
THE CHEMICAL INTERNET knowing paired with technological advancement within
the last century has catapulted yesterday’s discarded
Imagine marrying the intricacies of chemistry with
natural products into today’s pharmaceutical reality.
the processing power of computers. Chemist Bartosz
Gryzbowski of Ulsan National Institute of Science
and Technology (South Korea) is the driving force

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A FOODIE’S FIRST-IN-CLASS FDA APPROVAL Life Science Training from Industry Experts

GMOS FINALLY SWIM UPSTREAM

AquaBounty’s genetically modified salmon will make
it to your dinner table in less time than it took to toil
through the FDA pipeline. Coined AquAdvantage
Salmon, it is the first genetically modified animal
available for human consumption thanks to an
approval last month. The two year wait time to grow
the first batch pales in comparison to the 20 year
application period undertaken by Massachusettes-
based AquaBounty.
Public concerns contributed to the reasoning behind the
lengthy and rigorous review time for the AquAdvantage
Salmon application. To quell the safety concerns, the Because of the faster growth cycle, the AquAdvantage
FDA and AquaBounty are releasing the entire data fish require 25% less feed than conventionally farmed
from the application to the public—something not fish. This reduces both the environmental impact and
routinely done for new food or drug applications due to the cost to the consumer when compared to other
confidentiality. So, let’s take a closer look at the science farmed fish, and eases the pressure on declining wild
and piece through the numbers to find out exactly salmon populations from overfishing.
what’s on the menu for GMO salmon.
SAFETY CONCERNS: ALLERGENS
SPAWNING SUPER SALMON
Increased allergen levels or introducing new allergens
Wild salmon live mainly in the ocean, swimming not seen in regular salmon are some of the biggest
up rivers to their freshwater birthplaces to spawn. concerns of the GMO variety. The first evaluations
Farmed salmon, or salmon aquaculture, use open for allergenicity tested the Chinook salmon growth
cages in the ocean or self-contained systems to hormone for similarities to known allergens, with none
grow the fish, typically reaching maturity in about 30 found. The FDA analysis of the data related to known
months. AquAdvantage Salmon have been genetically salmon allergen levels was not significantly different
engineered to reach market size in a fraction of the from the levels of the allergens in non-GMO, farmed
time—only 18 months! salmon. In fact, increased levels of vitamin B6—that did
How do the fish manage to grow so quickly? First, not exceed the recommended daily allowances—were
researchers took a growth hormone gene from a salt the only significant differences between the two types
water relative—the Chinook salmon—and inserted it of fish.
into the AquAdvantage Salmon. Next, they engineered
the fish to produce the growth hormone year-round SAFETY CONCERNS: HORMONES
(instead of only in warmer months) by putting the For both AquAdvantage Salmon and non-GMO control
gene under the control of a promoter, or a type of “on” salmon, the levels of growth hormone were below the
switch. They borrowed the promoter from an eel known detection limit, and levels of estrogen and testosterone
as the ocean pout, which enables it to survive the did not differ significantly.
cold North Atlantic water by ensuring the continuous
The only potential cause for concern are the increased
production of so-called “antifreeze proteins.” When
levels of insulin-like growth factor (IGF-1) in the GMO
transferred into AquAdvantage Salmon, the promoter
salmon, at 10.26 nanograms (ng)/gram vs. 7.34 in the
signals the fish to produce growth hormone year-round.
control group. It is important, however, to put this

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number in perspective when compared to other animal chromosome, creating so-called triploid fish. This is
products that we eat. These numbers are only slightly a standard technique used for years to create sterile
higher that the levels of IGF-1 found in organic milk. fish for fish farms.
Beef from cows with no growth hormone treatments • P
 hysical: AquaBounty has taken significant steps
contain 275 ng IGF-1/ per ml of blood. The comparisons to ensure the physical containment of the GMO
suggest consumption of normal amounts of salmon. The fish eggs are produced at a facility in
AquAdvantage Salmon would not result in significantly Canada, and any eggs that pass through filters and
higher levels of dietary IGF-1 than diets containing into the drainage area will be killed by chlorine. The
regular animal products. salmon themselves are grown in large land-based
It is also important to remember that dietary IGF-1 likely tanks in Panama, with multiple filters and nets
does not impact human health. First and foremost, all installed to reduce the likelihood of escape to less
food proteins are broken down in the digestive track, than 1%.
meaning no individual would absorb an intact hormone • Environmental: In the very unlikely event that
from the foods they eat. And even if absorbed, the salmon eggs or salmon did escape production
structure of salmon IGF-1 is different enough from that facilities, environmental containment measures
of human IGF-1 that it is very unlikely to interact with would make it difficult for the fish to survive. For
the human IGF-1 receptor and promote cell growth. example, high salt concentration in water near the
production facility would kill any escaped eggs
THE ENVIRONMENTAL IMPACT while warm water temperatures near the Panama-
The other major public concern surrounding GMO based facility would be fatal to the salmon.
salmon is the possibility of escape and interbreeding
Even though retailers such as Whole Foods,
with wild salmon. Multiple redundant safeguards—
Costco, Target and Safeway are refusing to stock
biological, physical, and environmental—have been put
AquAdvantage Salmon on their shelves, there is still
in place to prevent the threat:
plenty of time to examine the data before the first batch
• Biological: The fish are sterilized, therefore arrives in two years. Regardless of whether the next
unable to breed with wild salmon. Sterilization FDA approved genetically modified animal for human
is accomplished by subjecting the eggs to consumption is a pig or chicken, the time has come for
pressure, which results in an extra copy of every the public to learn the science behind the headlines.

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CATCHING THE FIRST DROP OF CANCER Life Science Training from Industry Experts

THE LATEST IN CANCER DIAGNOSTICS classification of breast and prostate cancer, and is
developing cfDNA-based tests for breast (blood sample)
Hearing the words “it might be cancer” paired with your
and bladder (urine sample) cancers.
doctor’s perplexed look is enough to send shockwaves
through your body. Getting to the heart of a diagnosis Qiagen (Hilden, Germany) is developing cfDNA liquid
usually requires a surgical biopsy—removal and biopsy diagnostics in partnership with pharmaceutical
examination of the suspected tissue for visible signs companies such as AstraZeneca (London, U.K.), Tokai
of cancer. Pharmaceuticals (Boston, MA), Novartis (Basel,
Switzerland), and Eli Lilly (Indianapolis, IN).
Less invasive diagnostic tests—called liquid biopsies—
might just bring more choices to doctors and patients. EXTRACTING EXOSOMES
They are becoming today’s reality thanks to our
Exosomes are lipid-encased vesicles that contain
ability to isolate molecules from body fluids. These
cellular protein, DNA, and RNA and typically have
diagnostic innovations pair technology with the latest
surface proteins specific to their native cell. These
in biomarkers—and will be coming to an oncologist
attributes, combined with the fact that they are found
near you.
in many different body fluids, make exosomes a very
TERM OF THE WEEK: LIQUID BIOPSY attractive possibility for liquid biopsy. The idea is to
capture exosomes based on tumor-specific surface
Liquid biopsies provide information on the disease
markers or to collect exosomes and identify them as
status of cancer in patients through the detection and
cancer-associated by examining the enclosed DNA
analysis of biomarkers—think cell-free DNA, exosomes,
or RNA.
circulating tumor cells—found in body liquids such as
blood, urine, saliva and more. Hot new startup Codiak BioSciences (Cambridge,
MA) is looking at exosome-based pancreatic cancer
DISCOVERY BY CELL-FREE DNA diagnostics. Aptly named Exosome Diagnostics
(Cambridge, MA) is in clinical testing of exosome-based
When cells in the body die, they release cell-free DNA
urine and blood tests for prostate and lung cancer.
(cfDNA)—this includes dying tumor cells. cfDNA-
Rounding out the pack is Qiagen, who is eyeing the
based tests are a type of liquid biopsy because they
possibility of exosome-based cancer diagnostics to
seek out biomarkers in body fluids and then identify
compliment their work with cfDNA.
cancer-specific mutations using PCR or next-generation
sequencing analysis.
Trovagene (San Diego, CA) analyzes cfDNA found
in urine samples, which patients collect at home.
Currently, Trovagene has tests detecting mutations
associated with melanoma, colon cancer, and non-
small cell lung cancer, as well as the presence of viral
DNA for the diagnosis of human papilloma virus. Exact
Sciences Laboratories (Madison, WI) uses at-home
collection in their colon cancer test, which analyzes
cfDNA in stool samples for cancer-associated DNA.
Genomic Health (Redwood City, CA) currently markets
tissue-based genomic tests for the detection and

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CONSIDER CIRCULATING TUMOR CELLS Researchers at National University in Singapore
(Singapore) and MIT (Cambridge, MA) have developed a
The final category of liquid biopsy is perhaps the
microfluidics chip that routes cells from a blood sample
most obvious—circulating tumor cells (CTCs), or
into different channels based upon cell size. Although
cells splintered from a tumor and circulating in the
still in the preclinical research phase, this approach
bloodstream. The challenge lies in detecting CTCs: some
shows promise for capturing a wide range of CTCs.
estimates classify them as rare as one circulating tumor
cell per billion normal cells! Epic Sciences (San Diego, CA) adopts a “no cell left
behind” game plan thanks to technology developed
Janssen Diagnostics (Raritan, NJ) currently markets
by the Scripps Research Institute (La Jolla, CA).
CellSearch, the single FDA-approved test that allows
Automated fluorescence-microscopy identifies the
physicians to identify early CTCs from blood samples.
CTCs in blood samples placed on microscope slides.
Monoclonal antibodies (mAbs) capable of recognizing
A detailed analysis of three million cells per slide is
proteins on the surface of migrating tumor cells are
performed, each blood sample yielding approximately
chemically linked to magnetic nanoparticles and then
twelve slides. This technology may potentially hone in
added to a patient’s blood sample. These tumor-
on the presence of a single CTC. Epic Sciences currently
specific mAbs grab hold of the CTCs, and a strong
uses their test to perform analyses for biotech,
magnetic field is then applied to the sample, isolating
pharmaceutical, and clinical research partners with a
the captured cells for identification and analysis.
long-term goal of releasing a diagnostic product for
CellSearch is currently used to monitor the efficacy
reference labs.
of treatments for breast, prostate, and colorectal
cancer. A higher number of CTCs detected may Ultimately, the best liquid biopsies may contain a
indicate a higher incidence of metastasis, or a less than combination of all the above approaches. Biocept (San
effective treatment route if used to quantify cancer Diego) is leading the way by developing liquid biopsies
therapy success. that analyze both cfDNA and CTCs. Currently in the
process of commercializing a breast cancer test, Biocept
Another way to identify CTCs may be cell size—CTCs
also has its eye on combination liquid biopsies for both
tend to be significantly larger than other cells in the
colon cancer and melanoma.
blood, and this size differential may be exploited in
a microfluidics-based approach to cell separation.

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THE UBER OF THE HUMAN BODY? Life Science Training from Industry Experts

A TINY VESICLE WITH BIG POTENTIAL RNA-based therapies still struggle to achieve efficient
delivery mechanisms. Biologic drugs are too large to
Cambridge-based startup Codiak BioSciences made
enter cells on their own and might be able to catch a
headlines last month with $40M launch funding and
creatively crafted ride on an exosome. A few studies
another $40M if their technology shows promise.
have also suggested exosomes may be able to deliver
So, what’s the big deal? A tiny little particle—once
drugs across the elusive blood-brain barrier—another
described as a cellular trash truck—called the exosome.
common stumbling block.
First observed in the early 1980s, exosomes were
originally thought to be a way for cells to get rid of EXOSOMES TO THE RESCUE?
molecules they no longer needed, dumping their
Earlier this year, researchers at Temple University
contents into lysosomes. In the past decade, however, it
(Philadelphia, PA) demonstrated that in a mouse
has become increasingly clear that exosomes play a role
model of heart attack, damaged cardiac muscle could
in intercellular signaling because their routes and ability
be repaired by the injection of exosomes secreted
to transport cargo are much more than yesterday’s
by embryonic stem cells. Research at the University
garbage. Let’s dive into the exosome and find out
of New Mexico suggests that exosomes secreted by
why their potential as vehicles for drug delivery and
healthy cells near tumors can wipe out cancer cells—
therapeutics are the talk of the town.
avoiding collateral damage to nearby tissues often seen
TERM OF THE WEEK: EXOSOME in traditional therapies. This observation formed the
basis of the startup Exovita Biosciences (Albuquerque,
The term exosome derives from Greek roots, literally
NM). Codiak BioSciences is also eyeing exosome-
translating to “external body.” They are in fact very
based cancer treatments as well as exosome-based
small—about one one-hundredth the size of a typical
regenerative medicine.
human cell. These lipid-encased vesicles are secreted
from human cells and contain cellular protein, DNA, A BACK DOOR APPROACH
and RNA.
Cancer cells secrete exosomes containing proteins
CATCHING A RIDE and RNA specific to tumor antigens. The properties of
these newly earmarked cancer-derived exosomes could
Because of their established transportation procedures,
be back channeled into a cancer vaccine if Exocyte
exosomes may be able to deliver drugs to target cells.
Therapeutics (Singapore) fulfills their dream. Currently
How? Exosomes can be collected from cells grown in the
in preclinical studies, the putative vaccine works by
lab and “loaded up” with therapeutics via methods such
collecting exosomes excreted by tumor cells, incubating
as electroporation—pulsing the exosome with a short
those exosomes with white blood cells to stimulate
burst of electricity, creating temporary permeability
their tumor-fighting activity, and injecting the activated
which allows the drug to cross over its membrane.
blood cells back into the patient. Initial results suggest
Loaded exosomes would then deliver the drug to the
that exosomes are better at stimulating white blood
target cell based on target tissue surface proteins,
cells than isolated tumor antigens, and in most cases
similar to the way antibody therapeutics target specific
much easier to collect from a patient than biopsy-
tissues. Unlike antibodies, however, the exosome can
derived antigens.
actually enter the target cell because both the exosome
and cell membrane are made up of lipid membranes. Exosomes are high-beaming with promise all over
When like meets like, they fuse. the map, but be sure to tune in next week to find out
how these tiny vesicles with big potential might just
Exosomal delivery may help fill in the blanks for
revolutionize the fields of diagnostics.
various types of established therapeutics unable
to find their target market. The burgeoning class of

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THE ABCS OF HEPATITIS Life Science Training from Industry Experts

HEPATITIS VIRUSES EXPLAINED

Curing disease is serious business. Lou Reed, the
infamous leader of the rock band Velvet Underground,
died in 2013 of complications from hepatitis C and liver
disease. Death is the most profound fear for people
living with hepatitis C, but with today’s treatments
patients can beat this disease. Thank you Gilead’s
(Foster City, CA) Harvoni or AbbVie’s (North Chicago, IL)
Viekira Pak!
The word “hepatitis” means liver inflammation and
while the most common culprit is the hepatitis virus,
liver inflammation can also be attributed to other
causes—think excess alcohol, autoimmune disorders,
and non-alcoholic fatty liver disease. Let’s take a closer screened for HCV. An estimated 3.2 million people
look at the story behind the different hepatitis viruses in the U.S. have chronic hepatitis C according to the
and the drugs vying their way to a cure. Centers for Disease Control, and liver disease due
to HCV infection is the leading indication for liver
THE ABCS—AND D & E transplants in the U.S.
All hepatitis viruses infect liver cells and cause • H
 epatitis D (HDV) is transmitted through the blood
inflammation, but there are important distinctions and only occurs if HBV is also present.
in how they are spread and the type of illness that
• H
 epatitis E (HEV) is transmitted by ingestion of
they cause:
contaminated food or water and is seen almost
• H
 epatitis A (HAV) is usually transmitted by ingestion exclusively in parts of the world without clean water
of contaminated food or water. HAV is almost supplies. HEV is usually an acute infection, although
always acute, meaning people may get very sick for it may become chronic in cases where the immune
a few weeks, but then they clear the virus and do system is compromised.
not suffer long-term liver inflammation.
• H
 epatitis B (HBV) requires direct exposure to body MANAGING CHRONIC HEPATITIS B
fluids, such as blood or semen. HBV can be either Chronic cases of HBV pose a risk of cirrhosis or liver
acute or chronic. 90% of cases that occur before the cancer. There is no cure for HBV, but there are a variety
age of five become chronic, while the majority of of treatments that inhibit the enzyme the virus uses
those that occur after age five are acute. Since 1972, to replicate itself, including Epivir (ViiV Healthcare;
all donated blood has been screened for HBV. Brentford, U.K.), Hepsera (Gilead), Tyzeka (Novartis;
• H
 epatitis C (HCV) requires direct exposure to blood Basel, Switzerland) and Baraclude (Bristol-Myers
and is typically spread by using contaminated Squibb; New York, New York). These drugs slow down
needle or syringes. Although some people clear the the progression and make the disease manageable, but
virus within six weeks of infection, the majority— do not get rid of the virus.
approximately 80%—develop chronic infections. Recombinant interferon can also be used to treat HBV.
Chronic infections may initially be asymptomatic, Interferon is an inflammatory cytokine, meaning it is
but if left untreated, can lead to serious problems a signaling molecule that helps activate the immune
such as cirrhosis (scarring) of the liver or liver system to fight the virus. Interferon does not rid the
cancer. Since 1992, all donated blood has been body of infection and has unpleasant side effects,

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including fatigue, headache, insomnia, gastrointestinal • P
 aritaprevir: inhibits the HCV protein, NS34A. NS34A
distress, and depression. job is to cut large proteins into smaller ones; these
smaller proteins are critically important in the HCV
BEATING HEPATITIS C lifecycle. No small viral proteins, no HCV.
The landscape for hepatitis C changed at the end of • O
 mbitsavir: shuts down the HCV protein, NS5A. The
2013, with the FDA approval of Gilead’s Solvadi. As exact mechanism of action of Ombitsavir is not
a “nucleotide analog polymerase inhibitor,” Solvadi known, but it is thought to bind the NS5A protein
inhibits the viral polymerase—the enzyme used by HCV and inhibit its role in the HCV lifecycle.
to replicate its genetic material (RNA). To make copies
of the viral RNA, the polymerase simply connects new VACCINES FOR A & B, SO WHY NOT C?
building blocks—nucleotides—together in the same There are several reasons why there are safe and
order as the existing viral RNA. Solvadi is structurally effective vaccines for hepatitis A and B, but not for HCV:
very similar to nucleotides found in nature, so the
• H
 CV virus is more genetically variable than the
polymerase will incorporate the drug into a growing
viruses that cause HAV and HBV.
RNA strand. However, Solvadi has been chemically
modified so that once incorporated, the polymerase • Vaccines must be tested in animals before they can be
is unable to add any additional nucleotides, thereby tested in humans. It is difficult to find an animal model
halting viral replication that can host the HCV virus other than chimpanzees—
testing is limited due to ethical and cost concerns.
In 2014, Gilead gained FDA approval for a second drug,
Recently developed mice with grafted human liver cells
Ledipasvir, which inhibits the HCV protein NS5A. The
are proving to be valuable models for the disease.
exact function of NS5A in the viral life cycle is not
known—it is thought to play a role in viral replication, • It is difficult to find people to enroll in HCV vaccine
assembly, and secretion. Sovaldi and Ledipasvir are now trials. A new vaccine requires researchers to vaccinate
marketed together as Harvoni, and the combo clears people who are at high risk of infection. In developed
hepatitis C from infected livers. countries, those at highest risk are those who inject
street drugs and this particular population tends
The end of 2014 brought another HCV breakthrough
to avoid health care, and can be difficult to track
therapy with the approval of Abbie’s Viekira Pak. Viekira
for follow-up studies. There is a much larger at-risk
Pak knocks out HCV with a three-part punch:
population in developing countries where the infection
• D
 asabuvir: changes the shape of the viral tends to spread through contaminated hospital
polymerase. Without its proper shape, the enzyme equipment. However, the conditions that put these
is ineffective and viral replication is inhibited. patients at risk also makes developing country hospitals
a poor environment to run clinical trials.

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CANCER IMMUNOTHERAPY GOES VIRAL Life Science Training from Industry Experts

ONCOLYTIC VIRUSES • Deletion of viral gene ICP34.5: This gene codes for a
MAKE THEIR DEBUT protein that enables the virus to replicate in human
cells by blocking a human protein known as PKR.
Does a virus engineered to harness the immune
PKR prevents viral replication and is less active
system to fight cancer sound like a clever idea? Amgen
in most tumor cells. This makes the virus able to
(Thousand Oaks, CA) certainly thinks so, because their
selectively replicate in tumor cells.
talimogene laherparepvec (T-Vec) recently earned an
FDA approval to fight inoperable melanoma recurrent • Deletion of viral gene ICP47: This gene codes for
after initial surgery. Oncolytic viruses—like T-Vec—have a viral protein that inhibits the immune response
the attention of both industry media and mainstream to the virus by turning off a process called antigen
news programs. This new class of therapy is an elegant presentation. Normally, one of the key ways the
“hack” of the immune system, so let’s explore the immune system “knows” to attack a virally-infected
science behind the latest drug approval fighting cancer. cell is by recognizing antigens—or fragments of viral
proteins—displayed on the infected cell’s surface.
AN OPPORTUNISTIC VIRUS IS MADE Turning this process off helps the virus evade the
immune system. Turning it back on prompts the
T-Vec is an oncolytic virus—an engineered virus that
immune system to attack virus-infected tumor cells.
infects and kills cancer cells. Oncolytic viruses are
created in the lab by genetically modifying existing • Activation of viral gene US11: This gene increases
viruses in two ways: viral replication in tumor cells.

• Making the virus safe by removing genes that allow • Insertion of a gene for the human protein GM-CSF:
the virus to spead disease. This gene activates the immune system and aids in
the overall immune response towards the tumor
• Engineering viral surface proteins, so the virus
triggered by viral infection.
recognizes and binds to the cell receptors of
cancerous cells, disregarding the healthy, non-
THE ONCOLYTIC BANDWAGON
cancerous cells.
T-Vec is currently being studied for possible use in other
The oncolytic virus follows the same life cycle as any
solid tumors, and Amgen is collaborating with Merck
virus—once inside the human body it hunts down,
(Kenilworth, NJ) and Roche (Basel, Switzerland) to test
attaches to, and enters its host cell. In this case, the
T-Vec in combination with their checkpoint inhibitor
host happens to be cancer cells! The virally infected
therapies. The potential of this treatment has also
cancer cells are destroyed via the process of cell
beckoned other companies to the oncolytic bandwagon:
lysis—as the oncolytic virus multiplies inside of the
cells, it causes the cancer cells to burst open and kills
them. Spewing from the burst cells are new infectious
particles that further target remaining tumor cells.
The presence of a replicating virus also activates the
patient’s immune response, so the cancerous area is
further attacked.

THE VECTOR OF T-VEC

Interestingly enough, the virus used in T-Vec is a
modified herpes simplex 1 virus. The key modifications
made to ensure safety and efficacy are:

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COCKTAIL FODDER: RENEGADE
VIRUSES OF THE PAST
The idea of using viruses to challenge cancer is
cutting-edge, 21st-century science—but the inkling of
a cancer-fighting virus was first observed more than
a century ago. In 1904, an editorial published in the
American Journal of Medical Science revealed a
spontaneous regression of cervical cancer occurred
after administration of a rabies vaccination. A few years
later, a similar phenomenon occurred: the remission
of lymphoma after a measles virus infection. Our
modern understanding of viruses at the molecular
level combined with our increased ability to manipulate
genetic material made this century-old idea a medical
reality of today.

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A BIG PAIN Life Science Training from Industry Experts

THE SCIENCE BEHIND OPIOIDS

President Obama’s recent plan to fight opioids
highlights the dark side of a class of treatments
serving a vital need. Opioid pain medications manage
the severe short-term or chronic pain of millions
of Americans. While these medications mitigate
needless suffering, joining forces are the government,
corporations, and medical community to battle against
opioid abuse and addiction.
Here at WEEKLY, we always wonder: what is the science
behind the headlines? So, let’s talk about how pain
medications work, the different types on the market
and the approaches to developing less addictive
versions of opioid drugs.

OPIOIDS VS. NSAIDS

There are two main categories of pain medications,
opioids and non-steroidal anti-inflammatory drugs
(NSAIDs). Although these two categories of drugs work
differently, they do share one thing in common: both
are derivatives of natural products. The NSAID Aspirin
(Bayer, Leverkusen, Germany) is a synthetic version
of an extract from willow tree bark, and opioids are delivering the message, resulting in pain relief—along
synthetic versions of opium and morphine, which come with a feeling of euphoria in some users.
from poppy flowers.
Aspirin works by inhibiting an enzyme called LESSENING THE PAIN
cyclooxyrgenase 1 (COX-1). Once stopped, COX-1 is Short term medical used of opioid pain killers rarely
no longer able to produce signaling molecules, called leads to addiction—when properly managed. Due to
prostaglandins and thromboxanes. Prostaglandins the euphoria-inducing effects of the drugs, long term
and thromboxanes have a wide variety of functions, regular use, or use in the absence of pain, may lead
including mediating aspects of inflammation (fever and to physical dependence and addiction. And because
swelling) as well as promoting neuronal response to regular use increases drug tolerance, higher doses are
pain. Other NSAIDs, such as ibuprofen and naproxen, required to achieve the same effect, leading abusers
also work by inhibiting COX-1 or its sister enzyme COX-2. to consume pain pills in unsafe ways such as crushing
Opioid pain medications, such as Purdue Pharma’s and snorting or injecting the pills. According to the
(Stamford, CT) Oxycontin and Endo Pharmaceuticals’ Centers for Disease Control, 44 Americans die every
(Malvern, PA) Percocet, work by binding to receptor day due to prescription painkiller overdose. At the same
proteins on the surface of cells in the central nervous time, chronic pain is also a serious problem, affecting
system—think brain and spinal cord. While the approximately 100 million U.S. adults, while millions of
central nervous system is tasked with relaying pain others suffer acute pain due to injury or surgery. The
signals, opioids decrease the excitability of nerve cells medical need for these drugs is very real despite the
dark side.

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The answer to developing less addictive drugs may antagonist” of the mu opioid receptor. Simply put, a
be found in a drug that blocks pain without inducing competitive antagonist binds the receptor without
euphoria. These new drugs will need a different activating it. Since naloxone doesn’t activate the
mechanism of action than traditional opioid drugs, receptor, it doesn’t have any pain-relieving or euphoria-
which bind to the mu receptors of cells inside the inducing qualities; rather, it prevents the opioid drugs
central nervous system. Cara Therapeutics (Shelton, from binding. It may also displace opioids that have
CT) is developing drugs that bind to a different type already bound the mu receptor, aiding in the stoppage
of opioid receptor, the kappa opioid receptor. These of an overdose.
receptors are present on sensory nerves outside of the
central nervous system. Preclinical studies suggest that ENDORPHINS, THE NATURAL OPIOIDS
targeting these receptors could be effective at reducing The physical and psychological benefits of aerobic
pain without driving addictive behaviors. exercise are well known, and regularly active
Researchers at Indiana University are testing the populations typically report feelings of calm and mild
pain management potential of a peptide—a small euphoria after moderate to intense activity. These
protein fragment—that binds to calcium channels in feelings are linked to the production of endorphins—
nerve cells. These channels control the flow of calcium chemicals produced naturally by the body that bind
through nerve cells and play a critical role in sending to and activate the mu opioid receptors. In fact,
the pain signal to the brain. Preliminary studies suggest the word endorphin comes from the marriage of
this new peptide is able to reduce the transmission of “endogenous morphine”.
excitatory signals without completely blocking calcium Not everyone experiences the “runner’s high”—pretty
transmission required for nerve function. much in the same way that not everyone experiences
euphoric feelings from pain medications. These
AN ANTIDOTE TO AN OVERDOSE differences may help to explain why some people enjoy
Overdosing can be fatal since respiratory failure occurs exercise and others don’t, and why some people get
at high blood concentration levels of opioids. If an addicted to opioids—while others can take them or
overdose is suspected, the individual should be treated leave them.
as quickly as possible with naloxone—a “competitive

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ET TU, IMMUNE SYSTEM? Life Science Training from Industry Experts

AUTOIMMUNE DISORDERS: AN IMMUNE SYSTEM GONE AWRY

A STORY OF BETRAYAL In healthy individuals, the immune system serves to
“Et tu, Brute?” protect the body from foreign invaders such as viruses
The famous line spoken by Shakespeare’s Julius Caesar and bacteria. The immune response is driven by various
is synonymous with unexpected betrayal by a close types of white blood cells (WBC) that become activated
friend. A once trusted member of the inner circle, when they recognize foreign antigens—typically
Marcus Brutus joined the coup and turned on Caesar. proteins on the surface of viruses or bacteria. Once
The 23.5 million Americans suffering from autoimmune triggered, WBCs release signaling molecules (called
disorders also have a Brutus in their midst—their own cytokines) that fan out and mobilize other white blood
immune system. Type 1 diabetics must contend with cells for the fight—a process known as the secondary
white blood cells attacking their pancreatic beta cells. immune response. In the case of a natural infection,
Multiple sclerosis patients also find their once trusty the secondary immune response is a great way of
immune systems go awry destroying the myelin sheaths amplifying the initial response and quickly destroying
surrounding nerves. the pathogen. Once the pathogen is destroyed, the
immune response shuts down. But if the “invader”
The roots of the immune system’s treachery are
is a normal part of the patient’s body—when WBCs
poorly understood, but are slowly being teased out.
misinterpret one’s own antigens as foreign—the
This WEEKLY looks at the science and treatments
immune response remains active because the signal is
being pursued.
constantly present.
The autoimmune therapies of today rely largely on
“turning down” the patient’s own immune response.
They typically work either by:
• Inhibiting the cytokines
• Stopping the interaction between white blood cells
and cytokines
• Actively targeting and destroying a percentage
of the white blood cells that misread normal cells
as foreign
These therapies can be tremendously beneficial
in relieving the symptoms of chronic autoimmune
disorders such as Crohn’s disease and rheumatoid
arthritis. The downside is they can also put patients at
increased risk for a range of infectious diseases since
the overall immune response is invariably less active.

IN DEVELOPMENT:
PADLOCK THERAPEUTICS
The quest to develop a treatment for autoimmune
disorders that does not blunt the overall immune
response is the mission of a new biotech company,
Padlock Therapeutics (Cambridge, MA).

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Padlock’s technology is based on the observation that Protein citrullination is associated with a number of
a specific post-translational modification—a change different autoimmune disorders, including rheumatoid
made to a protein after it is made—turns otherwise arthritis, lupus, and multiple sclerosis. In rheumatoid
innocuous proteins into antigens capable of activating arthritis, and possibly other disorders, high levels
the immune system. The modification is known as of citrullinated proteins can be detected prior to the
“citrullination,” and results in the removal of a positive onset of actual symptoms, suggesting a possible
charge from a portion of the protein. Since protein early detection and treatment tool. Increased protein
structure is determined in part by charge-charge citrullination is also associated with neurodegenerative
interactions, this removal can cause the modified disorders such as Alzheimer’s disease, suggesting
protein to change shapes—becoming antigenic and another potential therapeutic area for PAD inhibitors.
invoking the immune response. Inhibiting this process
may provide a way to treat autoimmune disorders EASILY CONFUSED: AUTOIMMUNE
without dialing down the immune system. VS. AUTOINFLAMMATORY
Padlock is in the process of characterizing different The terms “autoimmune” and “autoinflammatory” are
inhibitors of the enzyme that carry out the both used to describe disorders of the immune system.
citrullination process. The enzymes are called proteins- What’s the difference?
arginine deiminases, or PADs—the “Pad” in Padlock Autoimmune disease refers to an activation of the
Therapeutics. There are five different types of PAD specific immune response—T-cells or B-cells—by
enzymes, so deciphering which are the safest to inhibit recognition of a specific “autoantigen” or self-antigen.
will no doubt play a critical role in the drug development At least initially, symptoms of an autoimmune disorder
process. The company expects to move into clinical are observed primarily in the region of the activating
testing by 2017. antigen, although as the disease progresses, the active
T- or B-cells may lead to systemic inflammation.
PROTEIN CITRULLINATION EXPLAINED
Autoinflammatory disorders, on the other hand, are
Protein citrullination normally plays a role in regulating caused by the overactivity of the non-specific immune
the expression of genes. This is because citrullination system—all of the white blood cells other than T- and
of the histone proteins around which DNA is wrapped B-cells. Although this overactivity may sometimes
affects how tightly the DNA is attached to the histone be triggered by an infection, often there is no known
proteins, which in turn impacts their level of expression. cause. Autoinflammatory disorders are largely driven
In general, the more “tightly wrapped” a segment of by cytokine production, though both autoimmune and
DNA is, the less often it is expressed. autoinflammatory disorders are thought to have a
genetic component.

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IS GENETIC VARIETY THE SPICE OF LIFE? Life Science Training from Industry Experts

UNDERSTANDING BASIS OF EVOLUTION Some mutations are deleterious, damaging the function
of the protein encoded for by the mutated gene.
Take a look at the person closest to you. Compare the
However, many mutations turn out to be neutral and
color of their eyes, the texture of their hair, even the
have no effect on gene function. How is this possible?
complexion of their skin to your own. Do you notice
Only 1.5% of the human genome actually codes for
a lot of differences? Genetic variation accounts for
proteins—the rest of it is non-coding, known as junk
the dissimilarities we observe between individuals—
DNA. Most of the time that a mutation occurs, it will
seemingly trivial ones such as the differences in eye,
occur in these non-coding regions and not affect a
hair, and skin color; more profound (but not necessarily
protein’s structure or function. Mutations may also be
harmful) variances such as the differing blood groups A,
neutral due to the redundancy of the genetic code—
B, AB, and O; and some medically significant differences
sometimes slight changes in a gene sequence result in
such as susceptibilities to particular diseases and
the same protein being produced. In some cases, even
responses to various drugs.
if a mutation occurs in a protein-coding region of the
Since I am on the road teaching this week, we will learn genome, the same protein gets made.
about the basis of evolution with excerpts from our
Very rarely, mutations can be adaptive and have
book: The Biotech Primer: An Insider’s Guide to the
a beneficial effect on gene function, conferring
Science Driving the Biotech and Pharma Industries.
an advantage on the organism. This is the basis of
Find out why mutations are not only essential to genetic
Charles Darwin’s Natural Selection or “survival of the
variation, but also how they play a role in evolution.
fittest” theory. Evolution is the natural selection of
Genetic variations impeding survival and reproduction
beneficial changes.
are often wiped from the population, potentially leading
to crucial changes in areas like appearance and biology. Some mutations are passed on from one generation to
another, and some arise during an organism’s life span.
MUTATIONS: THE MAIN SPICE Most mutations occur in somatic cells; that is, cells
IN GENETIC VARIATION that are not gametes (eggs or sperm). These mutations
are called somatic mutations and are not passed on
The genetic differences we see today arose as a result
to children. But mutations that do occur in sperm or
of mutations in the DNA sequence. Mutations can occur
egg cells, germ line mutations, will be inherited. If
by different means, and the outcome for the organism
the mutation is so severe that an organism cannot
can vary. There are a number of mutations in each and
survive, that organism does not pass it on to the next
every one of your cells. This is not necessarily cause for
generation. Therefore, over time, deleterious mutations
alarm—not all mutations are harmful. Mutations can
exit the gene pool.
have various effects on the function of a gene and its
protein product.
MUTATIONS AT THE GENETIC LEVEL
A substitution, or point mutation, results when one base
is swapped out for another. If the DNA polymerase
enzyme—the enzyme that copies DNA—accidentally
places a C where a G should be during DNA replication,
the substitution will alter the recipe.
Sometimes DNA polymerase may skip over a base,
which is a deletion, or add an extra base to the
sequence, which is an insertion. If a mutation goes
unrepaired, it results in DNA sequence changes that
will then be copied, becoming permanent. Changes

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in the DNA sequence as a result of errors by the DNA Polygenic diseases are caused by the interactions of
polymerase during DNA replication are rare, but they many different genes. Polygenic diseases are more
do happen. This is understandable when you consider common than monogenic diseases and include cancer,
the fact that every time a cell divides, it must copy heart disease, Alzheimer’s disease, and Parkinson’s
all 3 billion base pairs of DNA in just a few hours! disease. Polygenic diseases often have susceptibility
Fortunately, the DNA polymerase also has proofreading genes associated with them that increase the likelihood
ability, so it is able to “catch” and correct most mistakes, of the person developing the disease, but do not
but occasionally a mutation goes unrepaired, resulting absolutely predict its development—the ultimate
in DNA sequence changes that will then be copied, disease outcome will depend on various other genes
becoming permanent. in the individual’s genome, as well as environmental
In addition to errors made during DNA replication, factors. An example of susceptibility genes is the
mutations can also result from environmental factors, association of breast cancer with the BRCA 1 and BRCA
such as radiation from the sun or x-rays or from 2 genes.
chemicals in cigarette smoke.
COCKTAIL FODDER: A FEW
MONOGENIC VS. POLYGENIC DISEASE BASE DIFFERENCES
Mutations play a large part in disease. In a monogenic Scientists estimate there is a single base difference
disease, changes in one gene cause the disease. per every 3000-5000 bases, which is what makes
Examples of monogenic diseases include sickle cell humans look so different from one another. In essence,
anemia, cystic fibrosis, and Huntington’s disease. all people are pretty much the same—genetically
speaking, of course—minus a base difference here
and there!

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THE SCIENCE BEHIND THE NOBEL PRIZE Life Science Training from Industry Experts

DRUG DISCOVERY VIA by fluid collection due to the parasite lodging in the
NATURAL PRODUCTS lymphatic system, which would normally flush out
excess fluids.
Half of the 2015 Nobel Prize in Medicine went to
Satoshi Omura and William C. Campbell for their work Avermectin works to treat both infections by wiping
leading to the discovery of the drug avermectin, used in out the worms—it interferes with their nervous system
the treatment of parasitic diseases. The other half went and muscular function, resulting in the death of
to Youyou Tu for her efforts in uncovering the ground- the parasites.
breaking malaria treatment artemisinin.
UNCOVERING ARTEMISININ
Avermectin and artemisinin were developed from
Youyou Tu began with a large-scale screen of traditional
natural products—a time-honored and relevant
herbal medicines in malaria-infected animals, leading
source for medicine. As our ability to isolate, screen,
her to the plant artemisia annua. She extracted the
and modify chemical compounds from plants, fungi,
active component, known as artemisinin.
and microbes has increased, so too has our ability to
identify novel compounds for potential use in medicine. Malaria is caused by infection with a single-celled
protozoan—the type of organism you might remember
In this WEEKLY, we will understand the infections and
looking at in a drop of pond water underneath the
treatments involved in this year’s Nobel prize, and
microscope in high school biology class. Transmitted by
discuss natural products chemistry and its relation to
mosquitoes, the parasite multiplies in the victim’s liver,
drug discovery.
and then goes on to infect red blood cells. When treated
UNEARTHING AVERMECTIN promptly, patients usually make a complete recovery.
Severe malaria, however, can progress very rapidly and
In the late 1970s, Satoshi Omura isolated new
cause death within hours or days. Malaria is mostly
strains of the soil bacterium Streptomyces. Because
found in tropical and sub-tropical regions.
of Omura’s ability to develop unique methods for
culturing bacterial strains in the lab, he was able to The exact mechanism of action for artemisinin is not
identify 50 promising new cultures. These cultures fully understood—it may form chemical breakdown
were subsequently acquired by William C. Campbell, products that directly damage the malaria protozoan.
who screened them and isolated a compound with
remarkable efficacy against parasites (in both animals
ON THE MARKET & IN DEVELOPMENT
and humans). Avermectin treats river blindness and Since the dawn of man, various medicines have been
lymphatic filariasis—both caused by parasitic worms. derived from natural products—a few successful and
many utter failures. Companies continue the tradition
The worm that causes river blindness spreads via the
of using the natural world as a source of potential new
bites of the blackfly (simulium species), which lives near
drugs, but with today’s technology they do so with a
fast-running rivers in tropical regions. Depending on
little more specificity.
where the larvae take root, symptoms of the disease
range from skin problems to blindness. The blackfly is PharmaMar (Madrid, Spain) specializes in identifying
found mostly in sub-Saharan Africa. anti-tumor activity in compounds isolated from
marine organisms. The company’s first approved
The worm that causes lymphatic filariasis spreads
product, Yondelis, was originally isolated from a marine
through mosquito bites, and it is more common in the
invertebrate and now is chemically synthesized.
subtropical regions of Africa, Asia, Central and South
Yondelis is approved to treat soft tissue sarcoma and
America, and Pacific Island nations. The most noticeable
ovarian cancer and does so by interfering with cell
symptom of infection is elephantitis—a significant
division. PharmaMar has a number of other marine-
swelling of the arms, legs, or genitals. This is caused

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 Another company of note in the natural products
arena is Warp Drive Bio (Cambridge, MA). Warp Drive
combines the centuries-old idea of turning to nature for
medicine with twenty-first-century technology: rapid,
whole-genome sequencing of microbes. The idea is
to mine the genomes of microbes for gene sequences
associated with natural products of medicinal value.
These genes or gene clusters can be directly expressed
and their products analyzed for medicinal value.

COCKTAIL FODDER: ASPIRIN

The most famous—and ancient—example of a natural
product used as a drug is the practice of treating pain
discovered compounds in clinical development for a and fever with willow tree bark. Ancient Sumerian
range of different cancers. stone tablets, dating 2000 B.C., referenced willow
NovoBiotech (Cambridge, MA) has developed a bark preparations as medicinal. The father of western
proprietary cell culture method that gives previously medicine, Hippocrates, recommended willow bark to
impossible-to-grow microbes, the ability to flourish ease pain and reduce fever. By the mid-1800s, chemists
in laboratory settings—think expanded screening had isolated the active ingredient responsible for the
opportunities. This opens the door to a whole pain relief: salicylic acid. This ingredient alone, however,
new world of potential discoveries upon which proved to be too irritating to patient’s stomachs to
NovoBiotech can capitalize. Currently, they have enjoy widespread use. Chemists at a small German dye
compounds in preclinical development for cancer and company, Friedrich Bayer & Company (now Bayer),
infectious disease. developed the less irritating derivative, acetylsalicylic
acid, which launched as aspirin in 1899.

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PRESENTING THE NEW CLASS OF
TRANSCRIPTIONAL THERAPEUTICS Life Science Training from Industry Experts

DNA: CODING VS. NON-CODING IN DEVELOPMENT:

The completion of the Human Genome Project in 2003 SYROS PHARMACEUTICALS
revealed a big surprise: up to 98% of the DNA making Syros Pharmaceuticals (Watertown, MA) was born out
up the human genome does not code for proteins! The of the discovery of so-called super-enhancers—regions
notion that parts of the genome were non-coding had of DNA that activate gene expression, but are 10 to 100
been circulating for several decades, but when the times longer than typical enhancer sequences. Though
actual percentage was confirmed it blew the industry’s less common in the genome, they manage to bind to a
mind. This discovery prompted the launch of a multi- larger percentage of proteins responsible for activating
year, international effort called Project ENCODE gene expression—suggesting super-enhancers play
(ENCylopedia Of DNA Elements), with a goal to decipher a key role in regulating genes that define and control
the role of these non-coding regions. cell identity.
It turns out most of the non-coding DNA plays a role After identifying these zones of DNA, the company
in regulating when and how often coding regions are founders went on to demonstrate a large percentage of
used. For cells to carry out their specialized function, the disease-associate mutations found in the regulome
specific genes must be “turned on” or expressed, while are contained in the super-enhancer regions.
others must be “turned off.” Defects in the regulation This discovery opens up the door for a new class of
of gene expression (making too much or not enough of therapeutics dubbed “transcriptional therapeutics.”
a particular protein, or making it at the wrong time) are The moniker is derived from the first step of gene
linked to a number of different diseases. In this WEEKLY, expression: transcription, or converting the information
we will examine some key concepts behind gene in DNA to RNA, which is then converted to the
regulation and its association with disease. encoded protein.

TERMS OF THE WEEK: The company is currently conducting preclinical trials

REGULOME & ENHANCERS on an inhibitor of the protein CDK7, which plays an
important role in activating the expression of genes
The regulome consists of the parts of the genome that required for cell division. Early work has shown small
do not directly code for proteins—these non-coding molecule inhibition of CDK7—which binds a super-
parts essentially regulate when the protein-coding enhancer associated with a gene driving blood cancer—
regions are used. resulted in reduced expression of that gene. The data
DNA that codes for regulatory RNA, such as microRNA, look most promising for leukemia and lymphoma.
is one component of the regulome. Other components In addition to cancer, the company plans to focus on
are DNA sequences called enhancers. Enhancer the role of super-enhancers and gene regulation in
sequences are recognized by proteins that activate the inflammatory disorders as well as genetic disorders of
expression of associated genes—in other words, they the central nervous system and kidney.
act to “turn on” an intended gene.
EASILY CONFUSED: GENETICS
THE REGULOME & DISEASE VS. GENOMICS
Genome-wide association studies, to identify common Genetics is the study of how traits are passed from
mutations in disease populations versus healthy one generation to the next, such as the BRCA1 gene
populations, have been a research focus for the past conferring an inherited risk of breast and ovarian
several years. Far from being confined to protein-coding cancer. Genomics is the study of gene sequences, gene
regions, it turns out most of these disease-associated expression, and of the interaction of genes with each
mutations—93% of single base changes or SNPs—are other and with regulatory elements.
actually contained within in the regulatory regions (the
regulome). Astonishing!

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BREAKING DOWN LUNG CANCER Life Science Training from Industry Experts

LUNG CANCER PRIMER genotoxic, meaning that they cause cancer by directly
interacting with and damaging DNA. If that DNA damage
The hit TV series Breaking Bad features anti-hero
occurs in a gene involved in regulating cell division,
Walter White, who starts out as a sympathetic
cancer may result. Non-genotoxic carcinogens have no
character: a mild-mannered high school chemistry
direct interaction with DNA, rather they disrupt cellular
teacher with a nagging cough that turns out to be
structures and change the rate of either cell division or
lung cancer. Money problems precipitated by costly
processes that increase the rate of genetic error.
treatments, poor insurance, and a modest salary push
him to start cooking up meth to ensure the financial Radon gas exposure can result in cancer because it is
security of his family. Spoiler alert: The treatments radioactive, and the high-energy radioactive particles
succeed beyond his expectations, restoring his health given off as the gas decays can cause direct damage to
long enough for him to become an unexpected cellular DNA. Radon gas is released from the normal
meth kingpin. decay of radioactive elements occurring naturally in
soil and rocks. Radon is not considered dangerous
Breaking Bad is a fictionally extreme example of the
because it is usually present at very low levels. However,
chaos that can arise from a lung cancer diagnosis. In
it can sometimes build up to dangerous levels in well-
fact, lung cancer is the leading cause of cancer-related
insulated, tightly-sealed homes built on soil rich in
deaths in the United States. Let’s take a closer look at
uranium, thorium, or radium.
the molecular causes, the different types, and some of
the treatments in the clinic and on the market. Asbestos used to be a common insulating material
used in buildings and ships. The microscopic fibers
THE DANGER in asbestos can be inhaled and become lodged in
lung cells, triggering the activation of inflammatory
While Walter White did not smoke cigarettes, 90%
pathways that result in the release of mutagens and
of those affected by lung cancer are smokers.
factors that promote tumor growth. Since its hazards
Other causes of lung cancer include environmental
became well-documented in the mid-1970s, it is no
or workplace exposure to carcinogens (known
longer used as insulation.
cancer-causing agents) such as radon, asbestos, or
air pollution. In addition to carcinogen exposure, there are likely
genetic elements that make certain individuals more
Smoking causes cancer because the inhaled smoke
or less susceptible to lung cancer. Even though 90% of
contains a range of chemicals, 70 of which are known
lung cancer cases are caused by smoking, only about
to be carcinogens, including benzene, formaldehyde,
10% of smokers get lung cancer. In African-American
methanol, and acetylene. Some carcinogens are
populations, even when differences in smoking rates
and access to healthcare are controlled for, the rates of
lung cancer are higher. Both of these scenarios suggests
that there may be genetic factors that make certain
people more (or less) susceptible.

SMALL CELL
About 10% of lung cancer is small cell, meaning it occurs
in the very small cells found in the bronchii—the tubes
that branch off of the trachea, enter the lungs, and
divide into even smaller branches within the air sac.
There are currently no targeted therapies available
for small cell lung cancer, with chemotherapy and/or

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radiation as the main line of treatment. Broad ranging Ingelheim, Germany). Asians are much more likely than
therapies that harness the immune system are in other races to carry an EGFR mutation.
the pipeline—Merck’s (Kenilworth, NJ) Keytruda in About 5% of NSLC cases are caused by mutations in a
conjunction with chemotherapy, known as PembroPlus, gene known as anaplastic lymphoma kinase (ALK). ALK
is in Phase II. proteins activate cell division, and mutated versions
can drive cell division inappropriately. The drugs Xalkori
NON-SMALL CELL (Pfizer; New York City, NY) and Zykadia (Novartis;
Cancer that occurs within any cell outside of small cells Basel, Switzerland) inhibit ALK.
is referred to as non-small cell lung cancer (NSCLC),
The checkpoint inhibitor drug Opdivo (Bristol-Myers
making up the majority (~90%) of lung cancer cases.
Squibb; New York City, NY) has also been approved
A number of drugs targeting new blood vessel growth— for NSCLC patients whose cancers start growing again
angiogenesis inhibitors—have been approved for the after chemotherapy.
treatment of NSCLC. These include Avastin (Genentech;
South San Francisco, CA) and Cyramza (Eli Lilly; COCKTAIL FODDER:
Indianapolis, Indiana). WALTER’S DIAGNOSIS
There are also drugs that target specific NSCLC– Diagnosed with NSCLC, Walter White specifically had
associated mutations. For example, between 10% and an inoperable stage 3A adenocarcinoma. This means
35% of NSCLC cases are caused by the over-expression the cancer was initiated in the mucus-producing cells of
of the growth factor receptor EGFR. These types of the lungs and had spread to the lymph nodes (or other
NSCLC—more common in non-smokers—can be treated sites near the lungs), but had not spread to distant sites
by drugs that target and inhibit this receptor. These within the body. Some types of adenocarcinomas are
include Iressa (Astra Zeneca; London, UK), Vectibix caused by ALK mutations, so it is possible that Walter’s
(Amgen; Thousand Oaks, CA), Tarceva (Roche; Basel, miraculous recovery was caused by one of the ALK
Switzerland), and Afatinib (Boehringer Ingelheim; inhibitors discussed above.

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FEEDING CANCER Life Science Training from Industry Experts

THE METABOLICS OF CANCER carcinoma. The company is conducting Phase I trials of

CB-839 in both solid and hematological tumors.
Tackling cancer requires a multi-pronged strategy.
Currently, many of the cancer drugs on the market ARGININE, ANYONE?
work by inhibiting the signaling pathways that activate
Immunotherapies—therapies that activate a
cell division. These pathways are often the root cause
patient’s immune system to fight cancer—are
of cancer—a mutated gene leads to a dysfunctional
increasingly popular. They are viewed as a way to
signaling protein which tells the cell to divide
effectively fight cancer with fewer side effects than
inappropriately. In these cases, the cancer may be
traditional chemotherapies.
treated by inhibiting the dysfunctional protein.
T-cells—the “warrior cells” of the immune system—
Nevertheless, selectively targeting the metabolic
can be very effective partners in immunotherapy.
changes that cancer cells undergo is one prong worth
The highly effective checkpoint inhibitor and CAR-T
examining. In this WEEKLY, we’ll look at some of these
therapies activate cancer-fighting T-cells.
metabolic approaches, and discover another avenue to
target cancer. T-cells, like cancer cells, also require specific nutrients
to function optimally. T-cells (and other white blood
TERM OF WEEK: METABOLOME cells) need the amino acid arginine to be fully active. In
The metabolome is the complete set of small molecule some tumors, the enzyme arginase (which breaks down
chemicals found within a biological sample—think a arginine) is overactive. Calithera’s other drug candidate,
cell, tissue, organ, or organism. These small molecules CB-1158, works to inhibit arginase, thereby increasing
may be either the broken down products of normal arginine concentrations and helping to ensure that
metabolic reactions, or ingested from outside sources T-cells are fully active—and can battle with tumor cells.
(drugs, food additives, etc.). The tumor metabolome is Calithera is preparing to submit an investigational new
typically different from other tissues within the body, drug application to begin clinical studies in 2016.
and therefore may be a rich source of drug targets.
PLEASE PASS THE SUGAR
GOT SOME GLUTAMINE? Occasionally, people claim that sugar consumption
Cancer cells consume the amino acid glutamine at a in and of itself causes cancer. This is simply not true.
much higher rate than healthy cells. This dependence All cells, including healthy cells, require some sugar
is thought to arise from genetic changes in the cancer (glucose) to live—it’s where they get their energy from.
cells which alter fundamental metabolic pathways. Cancer cells grow and divide more rapidly than other
Preclinical research has shown cancer cells cultivated in cells in the body, which is why they consume excess
low glutamine conditions show a substantially reduced glucose once established. But simply removing sugar
amount of growth—a phenomenon sometimes referred from the diet will not prevent cancer. Cancer is caused
to as “addiction,” implying the cells are dependent by changes to the DNA of cells which interferes with
on glutamine. their ability to regulate cell growth and division. Once
a cell has become cancerous, reducing sugar in the diet
Calithera Biosciences (South San Francisco, CA) is
will not directly stop a tumor.
bringing that finding into the clinic with its experimental
drug CB-839, which inhibits the enzyme glutaminase, Too much sugar is unhealthy and may cause obesity—
the first step in the cancer cell’s utilization of glutamine. which is linked to increased risks for several types
Animal studies suggested CB-839 may show efficacy of cancers, as well as heart disease. Excessive sugar
towards triple negative breast cancer, non-small consumption is also a factor in developing type 2
cell lung cancer, multiple myeloma, and renal cell diabetes. So, avoiding excess sugar in the form of sweet

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snacks and processed food is good for your health, but
it is not a magic bullet for treating cancer.
Cancer cells do use glucose at a significantly higher
rate—as much as 200 times higher—than normal
cells. This metabolic abnormality could potentially be
exploited as a therapeutic target. Preclinical studies
suggest that therapies aimed at blocking cancer cells’
uptake or use of sugar may be therapeutically useful,
and researchers are working on ways to translate this
into clinical practice.

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THE POWERED EXOSKELETON Life Science Training from Industry Experts

THINKING OUTSIDE OF THE BODY gave Russian scientist Nicholas Yagn a patent for
an “apparatus for facilitating walking, running, and
Last WEEKLY’s focus on Duchenne muscular dystrophy
jumping”. The apparatus used compressed gas bags
got us to thinking, what products are available for
to augment user movements and required human
those with limited mobility? A new type of medical
power to operate. In the 1960s, General Electric and
device called a powered exoskeleton certainly caught
the U.S. military collaborated to produce the first true
our eye. Originally conceived as a tool to aid soldiers
exoskeleton—powered by hydraulics and electricity,
in lifting heavy objects, medical device companies
wearers could increase their strength by a factor
are turning to exoskeletons as a way to dramatically
of 25. At 1500 pounds, however, the suit had major
improve quality of life for a range of individuals. So, let’s
practical limitations and was never used in military or
uncork the current pipeline and learn about some of the
commercial applications.
inspirations behind these inventive products.
By the mid-2000s, leaps in electronics technology
TERM OF THE WEEK: and materials science had advanced to the point that
POWERED EXOSKELETON functional exoskeletons were a realistic possibility for
both military and medical applications.
An exoskeleton is an external skeleton thats supports
the body instead of (or in addition) to an internal
SOLID TAKES ON DMD
skeleton. Exoskeletons occur in nature—think insects
and crustaceans. Fact: turtles have both an internal and In 2012, investment banker Ilan Ganot’s son Eytani was
external skeleton—bones on the inside and a shell on diagnosed with Duchenne muscular dystrophy (DMD).
the outside. After learning more about the disease—such as the
majority of patients are confined to wheelchairs by age
A man made version of an exoskeleton is a mobile
11 or 12, and few live beyond their mid-20s—Ganot quit
machine consisting of an outer framework worn by
his job and relocated his family to Cambridge, MA to
a person. It is mechanized by a system of motors,
start Solid Biosciences.
hydraulics and/or pneumatics that delivers at least part
of the energy for limb movement, giving rise to the term Solid maintains a multi-pronged approach to discover
powered exoskeleton. treatments for DMD—including a preclinical research
collaboration with Pfizer (New York, NY) as well as as
A SKELETON IS BORN separate focus on gene therapy. Solid is also thinking
outside of the body with one of its key projects—
The idea of an exoskeleton to assist human activity
the Solid Suit, a powered exoskeleton device in
is not a new one. In 1890, the U.S. Patent Office
early development.
The Solid Suit is envisioned as a soft, wearable,
assistive device for DMD patients. The aim is to not
only assist DMD patients with daily activities, but to
preserve muscle function by helping to maintain at
least some muscle usage. Solid is partnering with SRI
International (Menlo Park, CA) to utilize innovations
derived from military technologies designed to offset
muscle fatigue and to augment muscle strength. In
addition, Solid is partnering with Parent Project
Muscular Dystrophy for critical patient feedback as the
prototype and clinical development moves forward.

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BIOM’S BIONIC LIMBS EKSO & REWALK
BiOM (Bedford, MA) is developing next-generation Ekso Bionics (Richmond, CA) markets Ekso, a wearable
bionic limbs. Founder Hugh Herr lost his own legs to bionic suit—also an exoskeleton—that enables
frost bite decades ago and has spent the intervening wheelchair users to stand and walk. The suit is coupled
years perfecting the technology to replace them. with crutches that contain sensors which send signals
BiOM limbs are covered with synthetic skin containing to a computer contained in the suit’s backpack. The
built in sensors, causing it to stiffen or soften as backpack also contains batteries powering tiny motors
appropriate with movement. They have been designed in the suit’s hip and knee joints, enabling movement.
by incorporating studies of normal human physiologies ReWalk’s (Marlborough, MA) Personal 6.0 System is a
to best mimic the natural movement of human limb similar product currently on the market.
muscle function, enabling propulsion very similar The next phase in exoskeleton development?
to what natural walking entails. The BiOM limb also Mind control. The 2014 World Cup launched with
interacts with the patient’s nervous system through a symbolic first kick by a paraplegic wearing an
electrodes implanted on the residual limb to which the exoskeleton. Developed by a team of neuroscientists
prosthetic is attached. at Duke University, the suit is controlled directly
BiOM is also developing exoskeletons for human by brain signals picked up by a cap worn on the
augmentation—in other words, exoskeletons to be patient’s head and relayed to a computer in the
worn by fully functioning adults, powering movement exoskeleton’s backpack.
and enabling the user to carry out tasks that normally
would be out of reach.

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DECIPHERING DMD Life Science Training from Industry Experts

THE ROOT OF DUCHENNE X-LINKED GENETICS

MUSCULAR DYSTROPHY DMD is almost exclusively seen in boys because the
World Duchenne Awareness Day called attention gene for dystrophin is located on the X chromosome.
to Duchenne Muscular Dystrophy (DMD) this past Females have two copies of the X chromosome,
Monday. Affecting one in 3,500 newborn baby boys whereas males have one X and one Y chromosome.
worldwide, this fatal disease is caused by a mutation in Because females have two copies of the dystrophin
the gene that codes for the dystrophin protein. Easily gene, it is unlikely that both Xs would have the
damaged muscle cells, progressive muscular weakness, dystrophin mutation and therefore, one of the Xs most
and serious medical problems—including significant likley produces a functional dystrophin protein—
impairment of the heart and lungs—are the direct ensuring she has healthy muscles.
results of this genetic mutation. Typical life expectancy The major down side for these females is she may pass
for a child with DMD is 25 years. the faulty copy on to male offspring, who have no “back
In this issue, we will discuss the root causes and up” copy of the gene and will be unable to produce
genetics of the disease, and find out the promising functional dystrophin.
therapeutic approaches in development. This form of inheritance gives rise to the term
“X-linked,” because the disease passed on via the
THE GLUE THAT HOLDS single X chromosome contained by a male offspring.
MUSCLE CELLS TOGETHER About two-thirds of DMD cases are caused by inherited
DMD patients produce no functional dystrophin mutations; the rest are the result of mutations that
protein. What does this protein do, and why is its loss arise spontaneously.
so debilitating?
In muscle cells (including cardiac muscles), dystrophin IN DEVELOPMENT: EXON SKIPPING
is part of a complex of proteins working together There is no cure for DMD at this time, but a number of
to strengthen muscle fibers—and to protect them companies are racing to develop treatments for the
from injury during the contraction and relaxation rare disorder.
cycles. Dystrophin serves to connect the proteins, So-called exon-skipping therapies are in development
which make up the muscle cell’s structural framework by Sarepta Therapeutics (Cambridge, MA) and
(the cytoskeleton), to their extra cellular matrix. The BioMarin (San Rafael, CA). These therapies use
extra cellular matrix is another network of proteins antisense technology to trick muscle cells into skipping
surrounding each cell. Because of these weblike over the mutated portion of the dystrophin gene when
connections, dystrophin is sometimes referred to as the dystrophin protein is produced. While the resulting
the “glue” that holds muscle cells together. Dystrophin protein is not fully functional, the production of a
is also thought to play a role in transmitting chemical partially functioning dystrophin protein appears to
signals within muscle cells. be enough to restore some functionality to damaged
Without functional dystrophin, muscle cells become muscle cells.
easily damaged and progressively weaker. DMD patients The term exon-skipping refers to gene structure—when
exhibit developmental delays early in childhood and a gene is converted to RNA, that RNA contains extra
experience progressive difficulties in standing and segments which are not used to make the final protein.
walking. Patients in their late teens and twenties often These excess segments are called introns and the
develop signs of cardiomyopathy or fatal respiratory high priority segments are referred to as exons. The
problems due to weakening of the muscles required exons are spliced together to form the final RNA that is
for respiration. converted to protein by the cell, essentially “skipping”
the unneeded introns.

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 IN DEVELOPMENT: GENE THERAPY
AND SMALL MOLECULE
While exon-skipping treatments seek to change the
expression of the mutated dystrophin gene, other
treatments in development seek to mitigate its impact.
Earlier this year, Milo Biotechnology (Cleveland,
OH) began clinical studies of a gene therapy based
approach to treating DMD. The company is using an
adenovirus-associated vector to deliver the gene for
the protein follistatin to patients. Follistatin increases
Introns are the segments in between the differently colored muscle growth in patients by inhibiting other signaling
exons shown above. Introns are discarded during splicing, molecules (such as myostatin) which restrain muscle
with the exons making up the final product.
growth. In addition to the current clinical study in DMD
by Milo, the therapy showed initial clinical safety and
efficacy in Becker muscular dystrophy—a less severe
form of the disease.
Santhera pharmaceuticals (Liestalv, Switzerland) has
completed Phase III clinical studies and is preparing
to file a new drug application for its small molecule
product idebenone. Idebenone works by increasing the
amount of ATP that muscle cells produce. ATP is the
form of chemical energy that cells can use directly to do
work. By amping up ATP production, the power switch is
turned on for the muscle cells.
The exon splicing shown above demonstrates how a
mutation is able to make it into the final protein. Another small molecule drug, Translarna, is currently
in Phase III —it is already approved by the EMA for
Exon-skipping therapies consist of an antisense use in Europe. Marketed by PTC Therapeutics (South
oligonucleotide—a synthetic strand of RNA or DNA Plainfield, NJ), Translarna works by helping the cell’s
whose sequence is complementary to the targeted protein-making machinery (ribosomes) continue to
sequence—which targets the mutated exon. This produce proteins even in the presence of a “nonsense
causes the mutated portion to be excluded from the mutation”—a mutation that would normally truncate
final protein. Sarepta’s exon-skipping treatment for protein production.
DMD is in Phase III clinical studies; BioMarin has filed a
The desire to find drugs that successfully manage—or
new drug application with the FDA.
even cure—this genetic disease are plenty high. With a
number of therapeutics in the DMD pipeline, WEEKLY
will be keeping an eye on the study results hoping for a
hero to emerge.

Exon skipping involves an “antisense oligo” shown above that

targets the mutation, rendering it incompatible. The mutation
is now excluded from the make up of the final protein.

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RNAI CRASHES THE PCSK9 PARTY Life Science Training from Industry Experts

NEWEST HOPEFUL IN CHOLESTEROL RNAI TECHNOLOGY EXPLAINED

LOWERING LANDSCAPE Recall from high school biology that RNA is single-
Just weeks after the biotech world celebrated the stranded (ssRNA) and is the set of instructions from the
approvals of two new cholesterol-lowering PCSK9 gene to the ribosome to make protein. Simply put, RNAi
inhibitors, Regeneron/Sanofi’s Praulent and Amgen’s technology creates a double-stranded RNA (dsRNA)
Repatha, a potential future rival arrived in style. by introducing an RNA sequence into a cell which is
Enter Alnylam (Cambridge, MA), with its RNAi-based complementary to the RNA for the particular gene to
experimental drug ALN-PSCsc, which just completed be silenced. When both ssRNAs meet, dsRNA is formed.
Phase I with positive results. The cell recognizes dsRNA as foreign and destroys it—
What makes PCSK9 inhibitors so hot, and why are the because dsRNA is seen as a “mistake.”
new cholesterol drugs on the market clutching their
pearls over ALN-PSCsc? Let’s find out why the PCSK9
and RNAi are creating a buzz in biotechnology.

ON THE MARKET: PCSK9

The body naturally keeps bad cholesterol in check
with low-density lipoprotein (LDL) receptors. These
receptors bind to excess LDL, which the liver cell
absorbs. The liver breaks down the cholesterol and
recycles the receptor back to the cell surface, where the
LDL receptor can bind to and remove more LDL.
PCSK9 is a protein that also binds to the LDL receptor,
which also triggers the liver cell to absorb the pair.
However, the entire complex is degraded and the
receptor is not recycled—sort of like a murder-suicide. When researchers introduce a dsRNA, it is processed
This results in fewer LDL receptors, impeding the by the cellular enzyme DICER and produces a “short
process of LDL removal. interfering RNA” , or siRNA. This siRNA then binds
a second enzymatic complex called RNA-induced
Repatha and Praluent are monoclonal antibodies
silencing complex, or RISC, which then recruits cellular
(mAb) that work by attaching to PCSK9, which prevents
mRNA whose sequence is complementary to the guide
the protein’s interaction with low-density lipoprotein
RNA. RISC then destroys both RNA strands—thereby
receptors on the surface of liver cells. By preventing the
preventing the production of the protein coded for by
degradation of these critical receptors, PCSK9 inhibitors
the targeted mRNA.
lower LDL levels and lessen the risk of a cardiovascular
event. Clinical results report lowering of LDL levels by The RNA-based therapeutics field has a history of failed
as much as 60%. One key advantage of the new PCSK9 candidates due to delivery problems—getting the
inhibitors is their safety profile—in clinical trials, the drug in the right place at the right concentration to be
adverse events observed were equivalent to that of therapeutically effective. The efficacy and durability
the placebo. of ALN-PCSsc is a significant accomplishment in the
RNAi world.

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IN DEVELOPMENT: ALN-PCSSC turn, binds a receptor on the surface of liver cells, called
the asialoglycoprotein receptor (ASGPR)—resulting in
Instead of inhibiting PCSK9 after its production in the
very efficient uptake of the RNAi. In order to increase
body (like the mAbs on the market), ALN-PSCsc blocks
stability, the RNAi has been modified to be resistant to
the production before it even starts. Early clinical study
cellular enzymes that would normally break it down,
results suggest that this is as effective as the mAb
making it possible for patients to one day be on a longer
therapeutics—and the efficacy is long-lasting, with
dosing schedule—potentially increasing compliance.
the potential for quarterly or even biannual dosing, as
With the buzz around ALN-PSCsc getting even louder, all
opposed to twice-monthly for Repatha and Praluent.
eyes will be on Alnylam every step of the way through
Alynylam’s approach was to create an RNAi molecule their clinical trials.
that targets the PCSK9 mRNA, and attaches the
synthetic RNAi to a sugar called “GalNac.” GalNac, in

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DODGING ANOTHER EBOLA OUTBREAK Life Science Training from Industry Experts

EBOLA FINALLY MEETS ITS MATCH

The headlining Ebola crisis of last summer devastated
West Africa, marking the largest outbreak since the
discovery of the disease in 1976. The glaring lack of
a treatment or vaccine caused the virus to quickly
spread within Guinea, Sierra Leone, and Nigeria. A small
number of infected healthcare workers were able to
receive an experimental antibody known as Zmapp
(Mapp Biopharmaceuticals, San Diego, CA) in the U.S
last year, with eight out of the ten surviving. Zmapp—
and other potential Ebola treatments—are now in
clinical trials, even as the outbreak has begun to wane.
Enter rVSV-ZEBOV (NewLink Genetics, Ames, IA;
Merck, Kenilworth, NJ), an Ebola vaccine that appears to
be both safe and highly effective. So, how exactly does (weakened). Attenuated vaccines may be more effective
the Ebola vaccine work? Let’s take a look into the world than killed vaccines at inducing a strong immune
of vaccines. response, because they more closely mimic a natural
infection. Common attenuated vaccines include the
TRAINING THE IMMUNE SYSTEM measles and mumps vaccines.

The principle behind vaccination is simple: by exposing •Pathogen subunits are vaccines that contain just one
an individual to an inactivated or otherwise harmless or a few proteins from the virus (or a toxin from a
form of an infectious agent, you can “train” their bacterium) in lieu of the whole pathogen. In some
immune system to recognize the pathogen by activating cases—as determined by clinical studies—these
memory B-cells and in some cases, memory T-cells. antigens alone are sufficient to induce an immune
response. A subunit vaccine on the market today is
If the person is subsequently exposed to the actual
Merck’s Gardasil against the human papilloma virus
pathogen, their immune system will immediately
(HPV).
spring into action, recognizing the old target and going
through their rehearsed game plan. Without vaccination •DNA vaccines are even smaller—they consist of a
or prior exposure to a particular pathogen, the immune segment of DNA (a gene) encoding a pathogen protein,
system can take several weeks to ramp up its response. and are used in place of injecting the whole pathogen
The speed at which Ebola is fatal underscores the or pathogen subunit. The idea is to induce a patient’s
need for a safe and effective vaccine to prevent such muscle cells to take up the plasmid and make the viral
devastating outbreaks in the first place. protein (antigen). While DNA vaccines have yet to hit
the market, many are effective at inducing an immune
FIGHTING THE GOOD FIGHT response in animals. One key challenge for this type
of vaccine lies in delivery—getting a patient’s muscle
While the premise behind all vaccines is similar—expose
cells to take up sufficient quantities of the antigen-
the patient to an inactivated pathogen to induce a
encoding DNA. Inovio Pharmaceuticals (San Diego,
protective immune response—the delivery platforms
CA) is developing an electroporation system that uses
vary considerably.
pulses of electricity to increase the uptake of injected
•Whole pathogen vaccines are made when the entire DNA vaccines, aiming to battle HPV, hepatitis C, HIV,
pathogen itself has been killed or attenuated and influenza.

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THE ONE-TWO-THREE RVSV-ZEBOV TESTING IN THE RING
PUNCH: RVSV-ZEBOV In order to get a new vaccine approved, manufacturers
The experimental Ebola virus vaccine, dubbed rVSV- must first demonstrate safety and efficacy. As with new
ZEBOV, combines elements of all three platforms drug approvals, this is typically a lengthy process, often
listed above. The prefix “rVSV” stands for recombinant taking up to a decade. Safety is first tested in healthy
vesicular stomatitis virus (VSV); ZEBOV indicates the volunteers, and then individuals at risk for contracting
Zaire strain of Ebola virus—the strain that caused the the infectious disease are given the vaccine candidate.
2014 outbreak. The at-risk individuals are followed for a period of
rVSV-ZEBOV was created using genetic engineering time—typically a few years—to see if they are less likely
techniques to splice the gene for the Ebola glycoprotein to contract the disease than those in the control group
into VSV. VSV normally infects cattle—so it is considered (who did not receive the vaccine).
safe to humans—but underwent further modification Since Ebola outbreaks tend to be infrequent and
to make it even safer. The only Ebola gene transferred is unpredictable, the traditional model of testing is
the glycoprotein, thus there is no chance of contracting difficult to complete. Merck used a “ring vaccination”
Ebola with the recombinant (genes from VSV and Ebola approach: anyone in contact with the Ebola victims
recombined) virus. received the vaccine as soon as contact had been
Why does the vaccine use the Ebola glycoprotein? This established—creating a “ring” of vaccinated people
protein, normally present on the surface of the virus, around one infected person. Another group of contacts
is used by Ebola to enter target cells in the human were not given the vaccine until three weeks after the
body. The glycoprotein binds to cells that line the blood initial exposure. Each group contained approximately
vessels, contributing to the massive hemorrhaging 2000 people. Among those given the vaccine
associated with Ebola infections. Using the glycoprotein immediately, no cases of Ebola were reported, whereas
as the vaccine antigen causes the patient to produce the control group reported 16 cases. The effectiveness
antibodies that bind to the glycoprotein, preventing it of the vaccine shows in the math, giving hope to
from infecting target cells and reducing hemorrhaging. perhaps one day controlling—or even eradicating—
Combining the Ebola glycoprotein with VSV closely the disease.
mimics a natural infection, so it does a very good job of
stimulating the immune system—without the attendant
dangers of an actual infection. VSV is easily eliminated
from the human body within a few days.

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STOPPING THE HEDGEHOG Life Science Training from Industry Experts

THE SCIENCE BEHIND THE • K

 aposi’s sarcoma (KS) is the uncontrolled growth
LATEST FDA APPROVAL of the cells that line blood vessels. KS is very
rare, typically seen only in those with severe and
Strong Phase II data launched Novartis’ (Basel,
prolonged immunosuppression. KS used to be a
Switzerland) Odomzo to an accelerated FDA approval
hallmark of HIV infection, but as HIV medications
for basal cell carcinoma last week. The company
have improved significantly, KS incidence
skipped Phase III testing for the skin cancer drug
has declined.
because 58% of patient’s tumors either shrank or
disappeared in Phase II. • Squamous cell carcinoma is a cancer of the epithelium,
which includes the top layer of skin as well as the
Odomzo is approved for patients whose basa l cell
cells that line the cavities, blood vessels, and organs
carcinoma has returned after surgery or radiation
throughout the body. Squamous cell carcinoma is less
therapy, or for those who are not candidates for the
common than BCC but more common than melanoma.
aforementioned treatments in the first place. Odomzo
About 4% of squamous cell cancers metastasize, making
is also attempting to tackle cancer in other areas—it is
it more likely to spread than BCC but less aggressive
in development for brain cancer as well as solid tumors.
than melanoma, in which the majority of patients
In this WEEKLY we will take a deeper look at the science experience metastasis.
behind Odomzo—and find out where a protein named
People with lighter skin tones are at higher risk for all
after Sonic The Hedgehog comes into play in the battle
types of skin cancer, since increased skin pigmentation
against skin cancer.
helps to block the damaging UV rays from penetrating
and damaging the DNA of skin cells. However, darker
THE SKINNY ON SKIN CANCER skin tones can and do get skin cancer, and thus should
A summer day at the beach or swimming pool isn’t also be vigilant about sun protection.
complete without a reminder to apply sunscreen.
The best way to prevent skin cancer is by minimizing TERM OF THE WEEK: SIGNAL
exposure to UV rays, so let’s understand the four main TRANSDUCTION PATHWAY
types of the disease:
A signaling molecule ignites the set of chemical
• B
 asal cell carcinoma (BCC) is one of the most reactions that activate a signal transduction pathway
common types of cancer and refers to uncontrolled in a cell—think of the process of a hormone attaching
growth of the skin’s basal cells, or the cells that to a cell surface receptor as an example. Signaling
line the deepest layer of the epidermis (the skin’s transduction pathways are typically cascades of
top layer). BCC is typically found on the head chemical reactions that eventually reach the target
or neck and can cause disfiguring growths—it molecule, such as the reactions within the cell created
rarely spreads to other tissues and is typically not when a hormone does its job successfully. Very often,
fatal. About 70% of cases occur on sun-exposed the net result of a signal transduction pathway is
parts of the body; the 30% of cases that are the “turning on” of specific genes—activating the
not the result of sun exposure are likely due to expression of their protein product.
genetic susceptibilities.
Signal transduction pathways can be highly complex
• M
 elanoma is the uncontrolled growth of the and interact with other signaling pathways within
pigment-producing cells known as melanocytes. the cell. Recognizing these pathways is critical to
Like basal cells, melanocytes are located in the interpreting the normal growth and development of
bottom layer of the epidermis. Melanoma is much cells, as well as understanding what has gone wrong in
less common than BCC, but is much more likely to disease states such as cancer.
spread to other tissues and be fatal.

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 ODOMZO THWARTS THE
HEDGEHOG PATHWAY
Odomzo is a small molecule inhibitor of the Hedgehog
Signaling Pathway, which plays an important role in
embryonic development. Defects in the pathway are
linked to various cancers, including BCC, small cell lung,
pancreatic, colon, prostate, and glioblastomas. This is
likely due to the fact the Hedgehog Pathway activates
the expression of genes affecting cellular proliferation
and survival—as well as angiogenesis (the development
of blood vessels). Odomzo inhibits a receptor protein
called Smoothened, which turns on the Hedgehog
Signaling Pathway after activation by another signaling
After binding to a receptor, the signaling molecule shown molecule—called Sonic Hedgehog. Yes, that’s right,
above sets off a cascade of reactions within a cell. Sonic Hedgehog.

COCKTAIL FODDER:
WHAT’S IN A NAME?
Have you ever wondered where some of the more
unusual gene or protein names come from? Originally
discovered in fruit flies, the Hedgehog Pathway
was so named because fly embryos deficient in the
protein were covered with small pointy projections—
resembling a hedgehog. The corresponding protein
in humans was dubbed “Sonic Hedgehog,” after
the popular 1990s video game character Sonic the
Hedgehog (Sega; Tokyo, Japan).

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A DROP OF BIOTECH Life Science Training from Industry Experts

BASICS OF BLOOD DISORDERS Phase I and Sangamo Biosciences (Richmond, CA), in

partnership with Biogen (Cambridge, MA), is currently
Blood is the carrier of a multitude of fundamental body
developing a genome editing treatment.
processes—responsible for delivering vital nutrients/
oxygen and for removing wastes. Like the highway DECIPHERING BLOOD CANCER
exchange feeding a city of life, blood is simply essential.
Like any other tissue in the body, the blood is
The branch of medicine concerned with the study,
susceptible to cancer. Blood cancers can affect any type
diagnosis, treatment, and prevention of blood related
of blood cell—white, red, or platelets—leading to their
diseases is known as hematology.
proliferation at the expense of the other blood cells.
This single therapeutic area covers a broad range of The three most common types of blood cancers are
diseases including hemoglobinapathies and blood leukemia (cancers of the circulating white blood cells),
cancers. Diagnosing and treating these varied disorders lymphoma (cancer of white blood cells within lymphoid
brings us many types of technologies. In this WEEKLY, tissue such as lymph nodes or spleen), and myeloma
we’ll gather some of the best biotech has to offer the (cancer of antibody-producing plasma cells)
field of hematology.
Most blood cancers are the result of an acquired, rather
THE HEMOGLOBIN PROTEIN than inherited, mutation in the bone marrow. The
exact cause of most blood cancers is not known, but
Hemoglobinapathies are the inherited disorders of
in some cases can be linked to radiation or chemical
hemoglobin, the all-important oxygen-transporting
exposure. Certain viral infections have also been linked
protein. Interestingly enough, there are only a few
to blood cancers, including Epstein-Barr virus (the virus
types of mutations that exist in the population because
that causes mononucleosis), human T-cell leukemia
hemoglobin is such a crucial protein. Anyone born with
virus, and human immunodeficiency virus. When these
a hemoglobin mutation most likely died shortly after
viruses infect white blood cells, they incorporate their
birth due to the marked devastation of protein function.
own genetic material into the host cell chromosomes
The most common type of hemoglobinopathy is sickle and occasionally disrupt genes involved in controlling
cell anemia. This disorder is caused by a mutation cell growth and division.
in the hemoglobin gene causing the hemoglobin to
clump together within the red blood cells, affecting the DIAGNOSING DISEASES OF THE BLOOD
overall shape of the cell. Normal red blood cells are Sickle cell anemia is diagnosed using a genetic test that
donut or disc shaped, while those affected are sickle detects the single base change in the hemoglobin gene
or “C” shaped. These sickle-shaped cells have trouble which causes the disease. Blood cancers, however, can
passing through the blood vessels—especially as they be caused by a variety of different mutations, making it
narrow at the branch points to feed into tissues. Vaso- difficult to develop a “one test fits all” type of scenario.
occlusive crises occur as the blood vessels become
Blood cancers are diagnosed by counting the number
clogged with improperly shaped red blood cells,
of each blood cell type, and determining if the amounts
blocking the flow of oxygen into tissues. Severe pain
fall outside of the normal range. A diagnosis can be
and potential permanent damage to the affected tissue
confirmed and further refined by looking at specific
are symptoms of sickle cell anemia, stemming from the
protein markers on the surface of blood cells. Blood
obstructed blood vessels.
cell counting and sorting is done via flow cytometry.
As a single-gene disorder, it is a prime candidate By suspending cells in a stream of fluid, and then
for emerging gene therapy and genome editing passing them through a “flow cell”—that allows only
technologies. Bluebird Bio’s (Cambridge, MA) one cell at a time to flow past a detector—the number
LentiGlobin is a gene therapy treatment for sickle cell in of each cell type can be determined using color coding.

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This process works by staining the blood cells with a
fluorescent-labeled antibody before they are loaded
into the flow cell. The fluorescent-labeled antibody
recognizes and binds to specific marker proteins on
the blood cell’s surface. The different marker proteins
indicate what type of cell it is, or in some cases, if it is
cancerous or not. As each cell passes by the detector, a
laser beam is directed toward the cell, and records the
presence or absence of a fluorescent label. Different
markers may be detected simultaneously by using
different colored fluorescent labels.

COCKTAIL FODDER: WHY IS

SICKLE CELL ANEMIA MORE
COMMON IN THE TROPICS?
parasite, however, the oxygen concentration within the
Sickle cell anemia just so happens to be more common
red blood cells drops—inducing hemoglobin clumping
in tropical regions, such as sub-Saharan Africa and
and the resultant red blood cell sickling. Sickled cells are
India. Enter the sickle cell trait—or, having just one copy
recognized as damaged by the spleen, and therefore
of the gene for defective hemoglobin—which serves as
destroyed and removed from circulation—inadvertently
a sort of protection measure against malaria.
also removing the malaria parasite. Having one copy
Since half of the hemoglobin produced by those with of the gene for sickle cell disease presents no serious
sickle cell trait is normal, their red blood cells do not problems, yet gives a distinct survival advantage against
sickle for the most part. When infected by the malaria the plethora of mosquitos in tropical populations.

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THE REALM OF NEXT GENERATION SEQUENCING Life Science Training from Industry Experts

BREAKING DOWN THE GENOME Hundreds of millions of DNA fragments are sequenced
simultaneously during NGS. The data from all of these
A billion dollar budget, a decade of work, and
fragments are processed to provide the final readout.
multinational collaboration brought the first human
genome sequence—via the Human Genome Project—
to the world at the turn of the century. Today, using so-
called “next-generation sequencing (NGS),” an individual
human genome sequence costs a mere $1,000 and
takes 24 hours. Even greater advances in cost saving
and time are expected in the next few years.
Full genome sequencing is revolutionizing medicine.
Current applications are focused on understanding the
genetic basis of cancers, and using that information
to develop better diagnostics and therapies to target
genetic subtypes. As the technology matures, we
can expect to see frequent applications in additional
therapeutic areas—even its use as a routine point-of-
care test.
The image above visualizes the steps involved
In this WEEKLY, we’ll break down NGS technology and in Next Generation Sequencing.

examine what is up next in the pipeline.

SEQUENCING BY SYNTHESIS: HI-SEQ
ONE DNA FRAGMENT AT A TIME
“Sequencing by synthesis” is a term applied to the
Today’s most successful NGS platforms work by taking most common methods of NGS, where the readout
many copies of an individual’s genome—usually 30— is established in an indirect fashion. Essentially, the
and breaking them up into overlapping fragments. sequence of each single-stranded DNA fragment is
Fragmentation is accomplished by mechanical force or determined by using it as a template to produce a
through the use of sonication (sound energy). second strand of DNA and detecting which nucleotide
Next, these individual fragments are attached to a base—A, C, G, or T—is incorporated at each position.
solid support, such as a microscale bead, and amplified Since we know that A always pair with T, and C always
(copied) many times. By the end of this amplification pair with G, by recording the order of the bases in the
step, multiple single-stranded copies of a particular newly synthesized strand we have also determined the
DNA fragment are attached to each bead. sequence of the template strand.

The sequencing machine is now ready to perform Illumina’s (San Diego, CA) Hi-Seq machine uses a
“massively parallel DNA sequencing.” The order of sequencing by synthesis method known as reversible
nucleotides—Adenine (A), Cytosine (C), Guanine (G), dye terminator. Simply put, Hi-Seq uses color coding to
and Thymine (T)—that make up each fragment will get a readout. For each round of sequencing, all four
be determined many times over. Once the individual nucleotides are washed over the template strand. Each
fragments are sequenced, the sequencing machine nucleotide has a specific fluorescent tag associated
determines the regions of fragment overlap, and with it—for example, each A may be fluorescent green;
from that information it pieces together the entire the C, fluorescent yellow, and so on. When the enzyme
genome sequence. that is synthesizing the new strand of DNA connects a
new nucleotide onto the growing strand, the machine
detects which color is incorporated, and correlates it to
a specific base.

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 T) is washed over the well in turn. If a given nucleotide
is incorporated, a hydrogen ion is released, and the
microchip records a change in pH. Whichever nucleotide
triggered a pH change is recorded as the next base in
the sequence.

IN THE PIPELINE: 3RD-GENERATION

The advances made in whole genome sequencing
have been truly impressive. However, 3rd-generation
sequencing is poised to revolutionize the field yet
again—with faster and less expensive read times on
even smaller machines.
Sometimes referred to as “single molecule
sequencing”—because it does not require sequencing
The illustration above shows how a color coded tagging by synthesis—3rd-generation technologies work by
system is used to determine the final readout.
directly “reading” the bases. One example is Oxford
Nanopore’s (Oxford, UK) platform, which passes a
SEQUENCING BY SYNTHESIS: single strand of DNA through a nanopore (or nano-
ION TORRENT scale hole) in a polymer membrane. The DNA strand
Another leading sequencing by synthesis platform is can only pass through each pore one nucleotide at
Thermo Fisher’s (Carlsbad, CA) Ion Torrent system. The a time; simultaneously, an electric current is also
name “Ion Torrent” signifies the systems reliance on passed through the membrane. The emergence of
measuring the release of a hydrogen ion, which occurs each nucleotide from the pore disrupts the current in
whenever DNA polymerase (the enzyme that makes the a characteristic way, depending on which nucleotide is
DNA strand) adds a new nucleotide to a growing strand passing through. This distinct disruption is noted by a
of DNA. This hydrogen ion release is detected as a slight detector, and the corresponding base is recorded.
lowering of pH. 3rd-generation sequencing is not yet commercially
How does this work in practice? To conduct Ion Torrent available, although Oxford Nanopore is conducting
sequencing, whole genomes are fragmented and pilot testing. Anticipated advantages of this technology
attached to microscale beads as described above. include faster read times, less expensive whole genome
However, these beads are placed in wells that contain sequencing, greater accuracy, and machines portable
pH-sensitive microchips. Each nucleotide (A, C, G, and enough to make point-of-care sequencing a realistic
possibility. Other companies working on 3rd-generation
sequencing platforms include Pacific Biosciences
(Menlo Park, CA) and Genapsys (Redwood City, CA).
The evolution of genome sequencing isn’t about to slow
down any time soon. Our healthcare system is only
as good as our technologies, and most believe that as
our DNA sequencing technology gets faster, stronger
and smarter so does our ability to diagnose and
treat disease.

The sequence is found by determining the change

in pH after a hydrogen ion is released.

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TARGETING THE CAUSE OF CYSTIC FIBROSIS Life Science Training from Industry Experts

BIOTECH CHANGES THE GAME mucus seen in the lungs of CF patients, making them
vulnerable to potentially fatal lung infections.
The FDA approval of Vertex Pharmaceutical’s (Boston,
MA) Orkambi last week further shifted the paradigm CF is an autosomal recessive disorder, meaning if an
of treating cystic fibrosis (CF) for up to half of its individual has one functioning copy of the CTFR gene,
sufferers—moving from a management of symptoms they are termed “carriers” and will not develop the
approach to targeting the underlying cause. Until disease. Two copies of the malfunctioning CTFR gene,
very recently, the only strategy against CF involved one from each parent, will equal a diagnosis. And while
reducing the risk of lung infections by taking mucus CF is always caused by a mutation, many possible
thinning medications and antibiotics—and rarely, mutation combinations have been associated with
late stage bilateral lung transplants. CF is an orphan the disease.
disease affecting around 70,000 people globally and is The mutations can be divided into two classes: those
prevalent in America, Europe and Australia. that lessen the quantity of CTFR proteins reaching the
This WEEKLY will discuss how Vertex’s two game- cell surface, and those that reduce the functioning of
changers—Kalydeco and Orkambi—target the root of the proteins reaching the cell surface. Drugs that work
the disease, and find out what’s next in line to take on by assisting CFTR to fold correctly and reach the cell-
Cystic Fibrosis. surface are referred to as CFTR correctors; drugs that
enable CFTR to function correctly once it has reached
CYSTIC FIBROSIS EXPLAINED the cell surface are CFTR potentiators.

CF is a genetic disease caused by one of several

TO THE MARKET: KALYDECO
possible mutations in the gene encoding the “cystic
& ORKAMBI
fibrosis transmembrane conductance regulator” (CTFR)
protein. The CTFR protein is critical for the production In 2012, Vertex Pharmaceuticals won FDA approval
of sweat, digestive fluids and mucus. CTFR is classified for Kalydeco, ushering in the first CF drug to treat the
as a channel protein—a category of proteins that create underlying cause of the disease. Kalydeco works by
a channel, or tunnel, across the cell membrane. This binding to the misfolded CTFR protein and increasing
specialized gateway allows things to pass through its ability to remain open and functional on cellular
the cell that will otherwise be denied entry or exit. surfaces—a CFTR potentiator. Although highly
Negatively charged chloride ions use CTFR to exit cells, effective, it is only a lifesaver for approximately 10%
and if CTFR is not functioning correctly, the chloride of CF patients. Kalydeco—by itself— is not helpful to
ions build up inside of cells. The buildup affects the fluid patients whose mutation causes CTFR to not reach the
balance of tissue, resulting in the characteristically thick cell surface.
When Vertex combined Kalydeco with newly developed
lumacaftor, a dynamic duo was born. Coined Orkambi,
the potential to benefit as many as 50% of CF patients
widened the eyes of the industry. The lumacaftor
portion targets the most common mutation (a single
amino acid deletion in the CTFR protein) responsible
for two-thirds of CF cases. In patients carrying this
mutation, the protein is so misfolded it never makes
it to the cell surface. Lumacaftor is a CFTR corrector,
it works by binding to and stabilizing at least some of
the misfolded proteins, improving their ability to travel
to the cell surface. Once there, Kalydeco kicks in and

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improves the function of CTFR, potentially bringing ON THE HORIZON:
positive outcomes for a big part of the CF community. SPYRYX BIOSCIENCES
CLINICAL TRIALS: Spyryx Bioscience (Durham, NC) is taking a different
CORBUS PHARMACEUTICALS approach to treating CF—a tactic which might also
benefit the entire patient population. Their efforts
Corbus Pharmaceuticals (Norwood, MA) is about to
are based on an observation of the link between the
take a swing at Vertex’s market share thanks to their
regulatory protein SPLUNC1 and the dehydration
current contender, Resonab, in Phase II clinical trials.
leading to thick mucus—the hallmark of CF. SPLUNC1
Rather than targeting the CFTR protein directly, Corbus
helps to modulate how much fluid is absorbed into lung
is treating CF as a chronic inflammatory disease. This is
cells. Spyryx is currently conducting preclinical testing
based on the observation that children as young as four
of inhalable peptide therapeutics (short segments
weeks old with CF have elevated levels of inflammatory
of proteins) based on the SPLUNC1 protein that will
markers, which further drives disease progression.
enhance the absorption of fluid into lung cells, relieving
Resonab is an oral anti-inflammatory—it works by the key symptoms of CF.
binding to and activating the CB2 receptor, which
The leap from simply managing CF to attacking its roots
is present on a variety of immune cells—think
shows the amazing potential of drug development.
macrophages, T-cells, and B-cells. Activation of the
With even more therapies in the pipeline, the ability to
CB2 receptor inhibits the immune function of these
fight CF on all levels is the path towards shutting out
cells, resulting in a resolution of the inflammation.
the disease.
Corbus is promoting the drug as a possible treatment
for all CF patients, not just those with a particular type
of mutation.

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WHAT THE HECK IS QRT-PCR DIAGNOSTICS? Life Science Training from Industry Experts

FROM IMPOSSIBLE TO POSSIBLE EASILY CONFUSED: GENE EXPRESSION

The genetics of cancer has progressed from the VS. PROTEIN EXPRESSION
impossible to a multifaceted mountain of possible. Gene expression occurs before protein expression. It
Comprehensive projects in whole genome sequencing is when the information from a gene is used to make
and tumor genome sequencing are providing data to messenger RNA (mRNA). Genomics scientists measure
unravel the genetic predispositions of cancer. the presence and abundance of mRNA in cells and
The other half of possible lies in quantitative PCR. tissues after gene expression.
This platform technology can identify types of Protein expression occurs after gene expression. It is
cancer, effective therapeutics, and aggressiveness the process after the gene has been transcribed into
of the disease from biopsy samples. Several leading mRNA. The mRNA is then translated into amino acids
diagnostics companies, including Genomic Health that join together to make polypeptide chains. These
(Redwood City, CA), Myriad (Salt Lake City, UT), chains fold to make proteins. Proteomics scientists
GenomeDx (San Diego, CA), Holigic (San Diego, CA), measure the presence and abundance of proteins in
and Biotheranostics (San Diego, CA) use variations of cells or tissues after protein expression.
quantitative PCR.
In this issue, we’ll take a closer look at how gene
THE BEAUTY OF HACKING INTO MRNA
expression is used to understand disease and develop One way to measure gene expression associated with
better diagnostics. disease is to determine how much of a specific mRNA
is present amongst the total cellular mRNA. This can
MRNA IS THE INTERMEDIARY be accomplished using a technique called quantitative
To develop diagnostics, scientists need to understand reverse transcriptase polymerase chain reaction (qRT-
how gene expression influences protein expression—or PCR).
how often a gene is translated into a protein. The first step in qRT-PCR is converting the isolated
In some cases, the actual sequence of a particular gene mRNA back into DNA. This is necessary because mRNA
does not differ between healthy and diseased—it is is highly unstable and it will not hang around long
simply expressed more (or less) often in the diseased enough to be analyzed. This is accomplished by mixing
population. In other cases, there may be a combination the isolated mRNAs with an enzyme called reverse
of gene mutation and expression. In fact, it is important transcriptase (RT) and adding adenine, cytosine,
to look not just at the expression of one gene, but at the guanine, and thymine (the nucleotide building blocks
ratio of expression between two different genes. of DNA). mRNA is copied back into DNA when the DNA
building blocks find their respective matches along
Messenger RNA (mRNA) is the intermediary between
the single-stranded mRNA sequence—same genetic
genes and proteins: a gene is transcribed into mRNA
information, but in the much more stable format of
and that mRNA is translated into chains of amino
DNA. These pieces of DNA are called cDNA because
acids—the building blocks of proteins. It is almost
they are copies of mRNA transcripts. For every mRNA
impossible to isolate the total amount of a particular
present in a sample there will be exactly one cDNA
protein made by a cell, whereas techniques for
produced. If we determine how many copies of a
isolating total mRNA content are in common practice,
particular type of cDNA are present, we will know how
making mRNA the perfect intermediary to determine
much of the corresponding mRNA was made by the
gene expression.
patient’s cells—and a diagnosis can be made.
To quantify the amount of a particular cDNA, a machine
called a thermocycler combined with the process of

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polymerase chain reaction (PCR) is used. PCR is a way COCKTAIL FODDER: WHERE
of making many copies of one specific DNA sequence. DOES RT COME FROM?
The final number of copies of that DNA sequence is in
The enzyme RT—which is used as the first step
proportion to the beginning number. By measuring the
in quantitative PCR to convert RNA to DNA—was
number of new DNA strands created from a specific
discovered in viruses called retroviruses, with HIV
cDNA, it is possible to back-calculate and determine the
being the most well-known example. Retrovirus literally
initial amount of that cDNA—thus the initial amount of
means “reverse virus” because they copy their genetic
mRNA present.
material (typically RNA) into DNA in order to incorporate
Quantitative PCR enables measurement of the amount itself into the host cell genome, working in a backwards
of a particular DNA produced by releasing fluorescent cycle to assimilate into their target.
signals as new strands of DNA are created. The more
fluorescence produced, the more copies of the cDNA
being measured are present. The technique is also
called Real-Time PCR because the fluorescent signal is
measured in real time—as the reaction is happening. If
we wait until the end to measure, it will be completely
saturated and not yield good data.

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POTENTIAL OF PCSK9 INHIBITORS Life Science Training from Industry Experts

$23 BILLION BLOCKBUSTER?

The PCSK9 inhibitor buzz keeps rolling, especially since
sitting before the FDA Advisory Committee earlier
this month. Amgen’s (Thousand Oaks, CA) Repatha
and Sanofi’s (Paris, France) Praluent are a new class of
cholesterol lowing drug looking to win final approvals
by late summer. Both companies are pioneers in the
realm of biologics (monoclonal antibodies) aimed at
the heart.
Early use restrictions may limit prescriptions to patients
with familial hypercholesterolemia—an inherited form
of high cholesterol that can be resistant to current good cholesterol because it transports the lipid
treatments. Doctor surveys suggest an interest in component in a compact fashion without losing
increasing access to a wider group of patients, leading or dropping the fat molecule when traveling in the
some analysts to predict a $23 billion market for arteries. HDL can even scoop up and expunge the LDL,
the drugs. or bad cholesterol. LDL is more fluffy and detaches
easily, wreaking havoc by oxidizing or subsequently
In this WEEKLY, we will suss out how PCSK9 inhibitors
attaching to arterial walls.
differ from their predecessor and what might be next in
line for managing cholesterol.
MECHANISM OF ACTION: PCSK9
MECHANISM OF ACTION: STATINS The body naturally keeps bad cholesterol in check
with low-density lipoprotein (LDL) receptors. These
The current standard of care for treating high
receptors bind to excess LDL, which the liver cell
cholesterol are small molecule drugs, called statins—
absorbs. The liver breaks down the cholesterol and
think Lipitor (Pfizer, New York, NY) or Crestor
recycles the receptor back to the cell surface, where the
(AstraZeneca, London, UK). These drugs work by
LDL receptor can bind to and remove more LDL.
inhibiting a liver enzyme called HMG-CoA reductase
inhibitor, which plays a key role in the synthesis of PCSK9 is a protein that also binds to the LDL receptor,
cholesterol. Less enzyme, less cholesterol produced. which also triggers the liver cell to absorb the pair.
However, the entire complex is degraded and the
About 70% of the cholesterol in a typical person’s body
receptor is not recycled—sort of like a murder-suicide.
is produced in the liver—which is why diet and exercise
This results in fewer LDL receptors, impeding the
may not always control high cholesterol. While statins
process of LDL removal.
are easy to swallow and work well for many patients,
they sometimes come with unwanted side effects, Repatha and Praluent work by attaching to PCSK9,
including muscle problems and an increased risk which prevents the protein’s interaction with low-
of diabetes. density lipoprotein receptors on the surface of liver
cells. By preventing the degradation of these critical
EASILY CONFUSED: HDL AND receptors, PCSK9 inhibitors lower LDL levels and lessen
LDL CHOLESTEROL the risk of a cardiovascular event. Clinical results report
lowering of LDL levels by as much as 60%. One key
Lipoproteins are divided into two categories: high-
advantage of the new PCSK9 inhibitors is their safety
density lipoproteins (HDL) and low-density lipoproteins
profile—in clinical trials, the adverse events observed
(LDL). A lipoprotein is a protein with fat molecules
were equivalent to that of the placebo.
attached for use as a transport system. HDL is called

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ON THE HORIZON year, Pfizer announced they are doing just that, with
clinical trials expected to begin in late 2015. Pfizer also
Therapeutic antibodies have a great track record for
has plans to develop a PCSK9 vaccine—essentially an
safety and efficacy and PCSK9 is a proven biologic
annual injection that would stimulate the patient’s
target in the cholesterol-lowering landscape. On the flip
immune system to develop its own naturally occurring
side, antibodies have the liability of a very expensive
antibodies against PCSK9.
production process which leads to higher prices for
patients. Oh... and therapeutic antibodies are delivered The outlook for treating high cholesterol is a little
via injection—which is not convenient when compared brighter. We at WEEKLY enjoy how the world of drug
to swallowing a statin pill. development always manages to have something else
up it’s sleeve—in the form of a pill, an injection and
The development of a small molecule inhibitor of
even a potential vaccine.
PCSK9 is a tantalizing proposition—and earlier this

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THE DANGER OF ANTIBIOTIC RESISTANCE Life Science Training from Industry Experts

THE STATE OF ANTIBIOTICS stops. Without a functional cell wall, bacteria die. Since
human cells do not have cell walls, they are not affected
One of the greatest public health challenges of the
by Dalvance and Orbactiv.
twenty first century—drug resistant bacteria—
prompted the first ever Antibiotics Summit at the White The third 2014 approval, Sivextra (Cubist, Lexington,
House earlier this month. Antibiotic resistance occurs MA), works by inhibiting bacterial ribosomes—the
when a few bacteria in a given population develop a enzyme that makes all bacterial proteins. Without new
genetic mutation that enables them to survive—even in protein production, the bacterium is unable to carry out
the presence of antibiotics. functions essential for life and dies as a result.

How do bacteria become drug resistant? Suppose a In February of this year, the FDA approved Avycaz
particular antibiotic works by inhibiting an enzyme (Actavis, Parsippany, NJ) to be used in combination with
required for bacterial replication. If one bacterium a previously approved antibiotic called cephalosporin.
mutates so the enzyme has a slightly different shape, Avycaz works by inhibiting the enzyme beta-lactamase,
the antibiotic is no longer effective. The mutated which bacteria secrete in order to break down other
bacterium lives on and continues to replicate, even as antibiotics—essentially boosting the potential of its
all the others die off. Over time, this resistant strain paired antibiotic.
becomes dominant, spreading from person to person,
remaining unchecked and thriving. The end result is
GROWING THE UNGROWABLE
multi-drug resistant bacteria. Although these new approvals are encouraging, none of
them work by entirely novel mechanisms. It is likely that
Antibiotic resistance is largely caused by antibiotic
resistance will eventually emerge—underscoring the
overuse—the more a bacterial population is exposed,
continued need to discover and develop antibiotics.
the greater the probability of mutations. This
begs the question: what is the current state of the Antibiotics have traditionally been found by screening
antibiotic economy? soil bacterium for compounds they release in order
to kill their competitors: other bacteria competing for
TO THE MARKET the same resources. A big problem with this method,
Last year saw the approval of three new antibiotics for however, lies in the fact that most soil bacteria do not
the treatment of acute bacterial skin and skin structure grow well in the lab—making it impossible to screen
infections, often caused by methicillin-resistant 99% of potential candidates.
staphylococcus aureus (MRSA). This year, an antibiotic
that enhances the effectiveness of its partner antibiotic
made it to the market.
Two of the 2014 approvals, Dalvance (Durata
Therpeutics, Chicago, IL) and Orbactiv (The Medicines
Company, Parsippany, NJ) work by inhibiting bacterial
cell wall synthesis. The cell wall is a layer of sugars and
amino acids (peptidoglycans) that surround bacterial
cell membranes, providing bacteria with structural
support, protection, and a filtering mechanism. The
new drugs are a synthetic lipoglycopeptide—a chemical
entity similar enough to the peptidoglycans so they
are easily incorporated into the cell wall, but different
enough so that once integrated, cell wall synthesis

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Earlier this year, a team of scientists at Northeastern CUTTING IT UP WITH CRISPR
University (Boston, MA) developed a method of
Another novel approach lies in the immune system of
cultivating bacteria in the lab by sandwiching them
the bacteria itself—the often buzzed about CRISPR/Cas
between layers of the soil separated by a semi-
9. CRISPR stands for “clustered regularly interspaced
permeable membrane. NovoBiotic Pharmaceuticals
short palindromic repeats.” These are short DNA
(Cambridge, MA) used the technique to screen 50,000
sequences that read the same forward and backwards
new types of soil bacteria and identified 25 new
and are found dispersed throughout the genomes of
drug candidates.
many types of bacteria.
NovoBiotic’s most promising candidate is Teixobactin, a
In the mid-2000s, scientists realized these sequences
compound that binds to two different lipid components
were a part of the bacterial immune system. Bacteria
of the bacterial cell wall, preventing them from being
insert bits of invading viral genomes between the
used to produce future cell walls. Neither lipid is rapidly
CRISPR sequences. If the bacteria is subsequently
evolving, so it is likely to take a much longer time to
infected by the same virus, the previously inserted viral
for resistance to come about. The compound is also
DNA is used to make RNA that recognizes and binds to
highly toxic to bacteria cells, a trait that should also
the invading viral genome—triggering the protein Cas9
defer resistance by quickly killing off target populations
to cut up the viral DNA.
before resistance emerges. Teixobactin may be the tip
of the screening iceberg. Researchers at Tel Aviv University (Tel Aviv, Israel)
and MIT (Cambridge, MA) are working to develop
SEQUENCING BY THE SEA CRISPR/Cas9 systems to target bacterially-encoded
Another large and mostly untapped source of potential antibiotic resistance genes. Their approach involves
new antibiotics—microbes of the ocean. Because of encoding CRISPR sequences on either side of an
significant environmental differences in temperature, antibiotic resistant gene, which may trigger the
salinity, and pressure between the ocean and the lab, gene’s destruction.
these microbes can be near impossible to grow outside The genes can potentially be delivered via viruses
of the marine environment. (called bacteriophages), which in turn can be
University of California (San Diego, CA) scientists engineered to target specific bacteria. This precision
are developing ways to circumvent this plight through technology, eliminates the issue of accidentally wiping
genomics. By isolating and sequencing the DNA of out “friendly” bacteria along with harmful bacteria—a
marine microbes, researchers identified gene clusters problem seen with many current antibiotics.
that they predicted would code for antibiotic-like While antibiotic resistance continues to grow, a
compounds. These genes were then transferred and several-pronged approach that includes a reduction
grown in bacteria, producing a compound dubbed in the unnecessary use of antibiotics, new initiatives
taromycin A. The result? Taromycin A impaired the to spur innovation in antibiotic development, and the
growth of several types of drug-resistant bacteria, development of more targeted diagnostics to guide
providing a potential platform for future antibiotics. their use are in order. Fortunately, the avenues to
discover new antibiotics appear to be widening, giving
hope for novel treatments in the near future.

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IMMUNE SYSTEM CHECKPOINT
THERAPIES ON THE CASE Life Science Training from Industry Experts

CANCER, MEET YOUR PD-1 AND PDL-1 STRAIGHT

NEWEST OPPONENT TO THE POINT
Immune checkpoint therapies received a lot of airtime Two other notable checkpoint proteins are PD-1 and
at American Society of Clinical Oncology (ASCO) PDL-1, the Dr. Watsons of this story. PD-1 is another
meeting in Chicago last week. The release of promising T-cell surface protein switch; its inhibitory activity
new clinical data has everyone buzzing about more is turned on when it meets the PDL-1 protein on the
cures for more cancers as a follow-on to last year’s race surface of host cells. Some cancers have increased
to the market. numbers of PDL-1 receptors, making these cancers
The hype seems to be justified. Simply put, these incognito experts.
new therapies thwart one of cancer’s most sly moves: In September 2014, the FDA granted accelerated
their ability to evade killer T-cells. This slight-of-hand approval to the PD-1 receptor inhibitor Keytruda
maneuver has limited a patient’s own T-cells to fight off developed by Merck (Kenilworth, NJ) to treat patients
tumors, and the efficacy of cancer vaccines/treatments with advanced melanoma. This approval was followed
that rely on a fully functioning immune system. This quickly by the December approval of Bristol-Myers
WEEKLY explains how immune checkpoint therapies Squibb’s (New York, NY) PD-1 inhibitor Opdivo, also for
get the job done. Read on and learn which of these advanced melanoma, and in March of this year for non-
therapies are on the market and which are barreling small cell lung cancer.
down the clinical pipeline. One potential advantage of PD-1 (or PD-L1 inhibitors)
over CTLA-4 inhibitors: PD-1/L1 inhibitors may have
TERM OF THE WEEK: IMMUNE fewer side effects. PD-1/L1 inhibitors seem to primarily
SYSTEM CHECKPOINT activate T-cells already present in tumor tissues. CTLA-
In order to prevent autoimmune disorders, our immune 4 inhibitor drugs have over-activated the immune
system has evolved “checkpoints”—proteins on immune response—resulting in autoimmune-like symptoms.
cells (such as T-cells) that need to be switched on or off
The talk of ASCO were the immune checkpoint therapies
in order to start the appropriate immune response.
in the pipeline:
Some cancer cells have cleverly evolved ways to exploit
these checkpoints and proliferate under the cover of
darkness. Immune system checkpoint therapies take
away the control of the switch before cancer has a
chance to flip it.

CHECKING OUT CTLA-4

CTLA-4 is a checkpoint protein on the surface of T-cells
that acts as an “off switch” for the sleuthing T-cells. It
signals them to remain in a resting state, kind of like
Sherlock Holmes asleep. AFTER THE CHECKPOINT
Bristol Myers Squibb’s (New York, NY) Yervoy is a Oncologists are excited about the new checkpoint
monoclonal antibody that targets the CTLA-4 proteins inhibitors for two main reasons:
and prevents cancer from hitting the T-cell off switch.
• They are proving to have longer-lasting efficacy,
Yervoy was the first checkpoint inhibitor drug approved
even compared to the highly effective, targeted
by the FDA in 2011 and is indicated for melanomas that
monoclonal antibodies. Once activated by
cannot be removed by surgery.

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 checkpoint therapy, the immune system can evolve benefit. Early studies indicate that genetic signatures of
and change with the cancer, unlike static therapies. tumors may also help predict who will respond well to
• They promise to fight a range of different cancers, PD-1 inhibitors. The diagnostics company NanoString
though their efficacy will vary between different Technologies (Seattle, WA) is developing a genetic test
subcategories of patients. to predict who is most likely to benefit from checkpoint
inhibitor therapies. Another possibility is simply looking
As with all medicines, physicians want to identify the
at tumor biopsies to determine if T-cells are already
patients most likely to respond to a particular drug.
present—increasing the likelihood that turning them
For example, the obvious criteria for giving a patient a
on will impact the tumor. With the buzz getting louder,
PD-1 inhibitor drug is the over expression of the PD-1
its only a matter of time before we see these intriguing
protein on the surface of his tumor cells—however, this
drugs take on the fight against cancer.
may not be the only way to identify patients who may

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A HEALTH CRISIS IN THE MAKING Life Science Training from Industry Experts

THE RACE AGAINST NASH Non-alcoholics with diabetes, obesity, and metabolic
syndrome run the risk of getting NAFLD and later
A silent epidemic creeping upon the Western world
on, NASH. It is estimated that approximately 25% of
pushed the headline grabbing acquisition of a Phase
Americans have NAFLD, creating a silent epidemic since
1 non-alcoholic steatohepatitis (NASH) drug last
symptoms often go unnoticed. Approximately 5% of
week. Boehringer Ingelheim (Ingelheim, Germany)
Americans are currently afflicted by NASH, with the
acquired Pharmaxis (Sydney, Autralia) due to the
numbers rising.
promise of PXS4728A. Several other companies also
have NASH drugs in development, such as Intercept, PHARMAXIS: PXS4728A
Genfit, Gilead, Galmed, Conatus, Raptor, and
Like many disease processes, inflammation is a
Galectin Therapeutics.
hallmark of NASH. As fat accumulates in the liver and
With no approved treatment on the market and liver overtaxes the organ, the release of inflammation-
transplant shortages only increasing, the race is on. inducing signaling molecules (called cytokines) set off
Let’s take a look at the science behind NASH and profile a cascade of events. Namely, the infiltration of white
the three of the contenders vying for a FDA approval. blood cells (such as macrophages) to the liver increase
the likelihood of cell death and scarring.
EASILY CONFUSED: NAFLD VS. NASH
Pharmaxis’ investigational therapy, PXS4728A, is a
Non-alcoholic fatty liver disease (NAFLD) happens when
small molecule inhibitor of the vascular adhesion
excess fat is deposited in the liver of people who drink
protein 1 (VAP1). Inhibiting VAP1 slows down the
very little to no alcohol. NASH is the severe form of
infiltration of white blood cells into the liver, thereby
NAFLD, and is characterized by liver inflammation and
limiting inflammatory damage. Boehringer Ingelheim’s
scarring. It is the number one cause of non-alcohol
investment in PXS4728A comes after encouraging
related cirrhosis, potentially leading to liver failure.
Phase 1 interim results.
NASH is the third leading cause of liver transplants in
the U.S. Pharmaxis is also considering PXS4728A as a potential
treatment for chronic obstructive pulmonary disease
(COPD), a disorder also associated with excessive
inflammation induced scarring.

INTERCEPT PHARMACEUTICALS:
OBETICHOLIC ACID
Recently granted breakthrough status by the FDA,
Intercept Pharmaceutical (San Diego) brings about a
promising small molecule drug called obeticholic acid.
Based on positive results from Phase 2 clinical studies,
Intercept announced last week the preparations to
initiate Phase 3 studies are under way.
Obeticholic acid works by binding to a so-called nuclear
receptor—a receptor protein that is present inside
of cells, rather than on their surface. When activated
by the appropriate signaling molecule, the nuclear
receptor moves inside the cell’s nucleus, where it binds
DNA at a specific location, activating the expression
of particular genes. Obeticholic acid specifically binds

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to and activates the nuclear receptor FXR. FXR then of NASH, the drug performed very well, preventing the
modulates the expression of a variety of genes involved onset and even reversing established NASH markers.
in lipid metabolism and glucose homeostasis that Surprisingly, recently reported Phase 2 clinical results
potentially disrupts the progression of NASH. failed to demonstrate notable improvement; however,
examining data from the more advanced NASH patients
GENFIT: GFT505 showed significant improvement. Genfit has recently
Yet another nuclear receptor activator is Genfit’s (Loos, announce plans for Phase 3 trials targeting only NASH
France) investigational GFT505, which targets the patients in more severe stages of the disease. With
receptors PPAR α/σ. Activation of these receptors turns the race to treat NASH only heating up, it remains to
on genes that increase the metabolism of fatty acids, be seen which contender will become the first FDA
resulting in a decrease of liver fat and improvement in approved treatment.
lipid profiles—as well as an increase in insulin sensitivity
and anti-inflammatory activities. In preclinical models

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ANTISENSE, RNAI AND MICRORNA EXPLAINED Life Science Training from Industry Experts

MAKE WAY FOR RNA BASED THERAPIES a key component of LDL cholesterol, to lower
cholesterol levels.
The up hill battle of RNA therapeutics to the clinic
continues despite extensive use in research. Recall from Over the past decade, the tumultuous love affair saw
high school biology that RNA translates DNA code into several antisense drugs fail due to lack of efficacy
a language ribosomes can understand in order to make and drug delivery problems. By targeting the right
proteins required by the cell. tissues (the most promising targets are in the liver) and
developing more stable formulations, Isis and others
Fighting the good fight are antisense, RNAi, and
expect to see more successes. Check out the antisense
microRNA. With their high specificity and relative
drugs in clinical development:
low manufacturing cost, these technologies may be
tomorrow’s biotech sweetheart. In fact, chances are
good that previously “undruggable” targets (that cannot
be accessed by small or large molecule drugs) are within
arm’s reach. However, the main hurdle continues to be
delivery—getting the RNA drug where it needs to be, in
high enough concentrations, to be effective.
This WEEKLY examines the similarities and differences
between the RNA therapeutics winding their way
through the clinic and into the marketplace.

PUTTING THE SENSE INTO ANTISENSE

The on again, off again love affair with antisense
therapeutics—that has been going on for over a
decade—was renewed in 2013 thanks to the FDA RNAI TO THE RESCUE
approval of Isis Pharmaceuticals’ (Carlsbad, CA)
Like antisense, RNAi takes advantage of naturally
antisense therapy Kynamro for the treatment of
occurring cellular pathways to target and destroy
familial hypercholesterolemia.
double-stranded RNA (dsRNA) to block the expression
Antisense drugs are short, synthetic pieces of nucleic of a disease-associated protein.
acid whose sequence is complementary to the mRNA
To activate the pathway, researchers introduce
that codes for a disease-associated protein. When
a double-stranded or “hairpin” shaped RNA. The
the antisense drug enters a patient’s cells, it binds to
enzyme DICER cuts up the hairpin to produce a “short
the disease-causing mRNA. This binding triggers an
interfering RNA” (siRNA). The siRNA binds to a second
enzyme called RNAse H to destroy the antisense-mRNA
enzyme called RNA-induced silencing complex (RISC),
duo (double-stranded RNA is seen as a mistake and
the siRNA/RISC complex then attaches to a disease-
destroyed). Without the mRNA, the disease-associated
associated mRNA. Now, double-stranded, RISC
protein simply is not made—stopping malignancy
destroys both RNA strands—siRNA and mRNA—and
in its tracks. Kynamro targets apolipoprotein B,
the disease is stopped. The RNAi therapeutics in clinical
development are listed below:

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 either makes too many or too few. If the patient is not
making enough of a particular miR, the therapeutic
approach is to deliver so-called “miR-mimetics”—
synthetic versions of the naturally-occurring miR in
diseases. The MicroRNA pipeline is summarized below:

MICRO RNA IN THE MIX

Antisense and RNAi are both synthesized in the lab
and delivered to patients to decrease the expression
of a disease-associated protein. MicroRNA (miR), on
the other hand, is a type of dsRNA made by cells to
REGULATING THE REGULOME
regulate gene expression. Like RNAi, miR is processed
by enzymes DICER and RISC into single-stranded RNA Enzymes such as RNAse H and DICER exist in our cells
capable of binding to disease-associated mRNA with a to regulate gene expression. Enter the regulome: the
complementary sequence. Since microRNAs only bind unknown world where the full complexity of these RNA
to one end of the mRNA, each miR is able to regulate disrupting pathways is beginning to be deciphered. Stay
multiple target mRNAs. tuned as RNA technology continues an avant-garde
approach to disbanding disease—a promising story
MiRs are noteworthy because their expression is
worth watching.
significantly altered in many disease states—the patient

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INCHING ACROSS THE BLOOD-BRAIN BARRIER Life Science Training from Industry Experts

A CONSTANT HURDLE IN This antibody takes advantage of a naturally occurring

DRUG DELIVERY process called receptor mediated transcytosis—or the
use of a receptor to transport something across the
When it comes to achieving success in drug
cell membrane.
development, picking the right drug target and
developing an effective inhibitor (or activator) is only Driving the bispecific antibody across the BBB is the
half the battle. transferrin receptor, which normally transports iron.
One arm of the BsA loosely binds to the transferrin
A drug candidate may appear promising in cell-based
receptor so it can easily be released once inside of
testing—and even in preclinical testing—but still fail
the brain. The other BsA arm binds to and inhibits a
to work in humans. It simply does not get to where
protein called beta-secretase, which is required for the
it needs to be in a high enough concentration to be
production of β-amyloid. β-amyloid is a protein thought
effective. This rings true for any number of drugs in
to play a role in the progression of Alzheimer’s.
development, but is most often the case with drugs
targeting sites within the brain. Brain Cancer

In this issue, we will focus on one of the biggest AngioChem (Montreal, Quebec) takes advantage of
technical challenges involved in drug delivery—getting a receptor called LRP-1, which naturally transports a
drugs across the blood-brain barrier. variety of proteins across the BBB. By analyzing the
range of proteins transported by LRP-1, AngioChem
TERM OF THE WEEK: BLOOD- came up with a sequence of 19 amino acids to attach to
BRAIN BARRIER and guide medicines to the LRP-1 transporter. Currently
in Phase II, the company is targeting paclitaxel, a potent
Tiny capillaries crisscross the brain to bring nutrients
anti-cancer drug to treat brain cancer.
to and waste away from the tissue. These capillaries
are lined with cells that fit so tightly together Parkinson’s
most substances—including 95% of all drugs—are Researchers at the University of North Carolina
prevented from making it over the border. This mostly (UNC) work with another natural process, called
impenetrable border is the blood-brain barrier (BBB). exosome-mediated delivery, to break through the BBB.
Some substances that do make it through the BBB Cells release exosomes, or nano-sized fat capsules,
are water, oxygen and certain hormones (such as to transfer material from one cell to another. In the
estrogen and testosterone) via diffusion. Glucose, the lab, these vesicles are isolated and loaded up with
all-important energy source of the brain, uses special a therapeutic payload. UNC researchers are using
transport proteins to shuttle itself into the brain. exosomes to deliver the anti-inflammatory enzyme
catalase (a Parkinson’s drug) to mice via a nasal spray.
BREACHING THE BBB Spinal Muscular Atrophy
Getting large therapeutic proteins past the BBB and AveXis (Dallas, TX) uses the adeno-associated virus
into the brain is one of the great drug development (AAV) to deliver SMN proteins to motor neurons in
challenges. Below are four different methods currently Spinal Muscular Atrophy (SMA) infants. SMA affects
being pursued: the motor nerve cells in the spinal cord, taking away
Alzheimer’s the ability to walk, eat, or breathe—it is the number
one genetic cause of death for infants. AAV serotype
There are many effective therapeutic antibodies,
9 is delivered intravascularly and then crosses into
however there is no mechanism for getting them into
the brain; it is the only serotype to cross the BBB in
the brain. Enter Genentech (South San Francisco, CA)
therapeutic doses. Currently in Phase I/II at Nationwide
who, still in the research phase, is testing a possible
solution in the form of a bispecific antibody (BsA).

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Hospital (Columbus, OH), it is the first clinical trial using
systemic gene therapy.

COCKTAIL FODDER: MULTIPLE

SCLEROSIS AND THE BBB
Part of the disease process of multiple sclerosis (MS) is
a breakdown of the blood-brain barrier, likely caused
in part by chronic inflammation. This breakdown is
what allows white blood cells (such as T-cells and
macrophages) to enter the brain and attack the myelin
sheath, the insulating lipid layer that surrounds
neurons. Certain biologics—like Biogen’s (Cambridge,
MA) Tysabri—normally are unable to cross an intact
BBB and actually take advantage of the breakdown to
penetrate the brain to treat MS.

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PACKING A PUNCH WITH THE IMMUNE SYSTEM Life Science Training from Industry Experts

THE INNER WORKINGS OF SPECIFIC IMMUNITY:

YOUR IMMUNE SYSTEM T-CELLS AND B-CELLS
What do monoclonal antibodies, CAR-T therapy, When non-specific defenses are unable to rid the
and 3D vaccines all have in common? They are body of an invading pathogen, it’s time to call for back
immunotherapies, or therapies that activate a patient’s up. Waiting for the call are T-cells and B-cells, which
own immune system to fight or prevent a disease. make up specific (or adaptive) immunity, ready to fight
While immune system activation can help save a life, foreign proteins.
an overactive immune system can potentially attack T-cells and B-cells are highly specialized to recognize
the body it is charged with protecting—the basis for unique targets, thanks to distinctly shaped receptors.
autoimmune disorders. Once a match is made, the receptor binds to the target
The biotech industry has elegantly hacked the immune and the T-cell or B- cell is activated.
system—a highly complex network of cells, tissues, T-cells attack infected cells floating around the body.
and signaling molecules—to make some of the leading These cells display foreign proteins on their surface and
immunotherapies, like Abbvie’s (North Chicago, when the T-cell encounters a non-self signal, it begins
IL) Humira. Let’s discover how the immune system its mission.
operates at the cellular level.
Activated T-cells divide rapidly and produce two types
NON-SPECIFIC IMMUNITY: of cells: killer T-cells and helper T-cells. Killer T-cells
MACROPHAGES AND NEUTROPHILS roam the body in search of their preprogrammed
target, and seal the deal by injecting granzyme B. In
Non-specific (or innate) immunity is the front line
contrast, helper T-cells do not actively target infected
defense against invading pathogens—think viruses and
cells, they release inflammatory cytokines and
bacteria. The troops defending against foreign invaders
activate antibody-producing B cells, killer T-cells, and
are specialized white blood cells (WBC). Most WBC in
macrophages to respond en masse. Helper T-cells are
the body are non-specific defenders, meaning they will
so critical that the immune system is crippled when
attack in the same fashion without stopping to consider
they unable to do their job—remember the human
the weakness of the enemy.
immunodeficiency virus (HIV) only infects helper T-cells.
Freely circulating in the bloodstream are macrophages
B-cells divide and alert other cells to destroy the
and when they encounter a foreign invader, they
pathogen once activated. They also clean up the
simply eat it. Macrophages recognize that a bacterium
battlefield and plan for future attacks instigated by
is foreign because its surface contains fats, proteins,
the same foreign invader. Most B-cells produce plasma
and carbohydrates that are different from those found
cells that secrete antibodies. Antibodies recognize
on human cells. Other types of non-specific defenders
and bind to any bacterium or virally-infected cell that
include neutrophils (which also recognize and engulf
bears an activating protein (antigen). The binding action
invaders) and natural killer cells, which inject the
triggers other immune cells, such as killer T-cells or
protein granzyme B into invaders, triggering cell death.
macrophages, to sweep in and destroy the invader
Non-specific defenders become activated by a threat attached to the antibody.
and release “inflammatory cytokines,” or signaling
Immunotherapies use concepts from specific immunity
molecules that activate other immune cells. The
to their advantage. Antibodies have been adapted for
inflammatory response is kicked into gear, ensuring a
use as therapeutic monoclonal antibodies and bispecific
rapid and comprehensive retaliation.
antibodies. Also, T-cells are engineered to recognize and
attack cancer cells in CAR-T therapy.

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THE WATCHMEN: DENDRITIC CELLS (Genentech, South San Francisco, CA)—work by
shutting down key parts of the response. Humira and
In order for T-cells to become fully activated, they
Enbrel inhibit the inflammatory cytokine TNF-alpha
must encounter the foreign invader in the context
and are approved for a range of inflammatory diseases.
of a “professional antigen-presenting cell,” called a
Rituxan, approved for rheumatoid arthritis, works by
dendritic cell. They are white blood cells that engulf a
reducing the number of B-cells that target the synovial
virus or bacterium, digest their protein components,
tissue of joints.
and “display” portions of those proteins on their
surface. Dendritic cells play a key role in developing The body has natural checks on the immune system
therapies, especially with Dendreon’s (Seattle, WA) 3D in order to prevent inflammatory disorders. A part
Vaccine Provenge. of the specific immune system development is a
screening process that terminates T-cells or B-cells that
THE OTHER SIDE OF THE mistakenly recognize the body’s own tissues. Even after
COIN: AUTOIMMUNITY passing the screening test, T-cells have certain protein
activators (called checkpoints) that must be turned
The immune system prevents us from falling deathly
on in order for the T-cell to become fully active. Some
ill as it responds to constant microbe exposure;
types of cancer cells exploit these immune checkpoints;
however, an overactive immune system can cause
however, checkpoint inhibitor therapies dismantle this
serious problems.
evasive mechanism used by cancer cells to stay hidden
When the immune system goes into overdrive, chronic from T-cells.
inflammatory disorders such as Crohn’s disease,
From one side of the coin to the other, the immune
rheumatoid arthritis, and psoriasis wreak their havoc,
system continues to amaze the industry as new
mistakenly attacking the body. White blood cells are
pathways and targets are discovered. A delicate
activated to target innocent cells in the body and
balance of the body’s toughest fighters, understanding
release inflammatory cytokines to sustain the response.
and optimizing the immune system is central to the
Biologic drugs that treat these disorders—like Humira, immunotherapy paradigm.
Enbrel (Amgen, Thousand Oaks, CA), and Rituxan

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READY. SET. CRISPR. Life Science Training from Industry Experts

CRISPR/CAS9 TECHNOLOGY a single base deletion may cause unintended

BUILDS UP STEAM consequences. If the repair occurs in the middle of
a gene, the minor error can be enough to disrupt
The world definitely weighed in after Chinese scientists
gene function and halt the production of the
published a paper detailing the use of CRISPR/Cas9
corresponding protein.
to edit nonviable human embryo genomes. While the
embryos used were never intended to become a living • Homology Directed Repair (HDR) relies on a highly
human being, this controversial milestone created a similar (homologous) DNA segment to repair the
firestorm of opinion. break—visualize the missing bridge section built
elsewhere and helicoptered in to fill the break.
Tweets, blogs, and mainstream news stories were abuzz
about the opening of Pandora’s box. Even Jennifer While these repair pathways are the body’s natural
Doudna, one of the creators of the technology, called way of fixing a break in the genome, the process has
for a moratorium. Just today, NIH stated they will not been hacked by scientists. Double-stranded breaks are
fund the use of embryos in genome-editing research. engineered to occur at specific locations, activating the
In the end, CRISPR/Cas9 did not successfully edit the intrinsic cell repair pathways of NHEJ and HDR.
embryo genomes as intended, which underscores the
juvenescence of the technology.
CRISPR/CAS9 ON THE RUN
The secret to good genome-editing lies in creating DSBs
The brave new world of genome-editing continues to
in therapeutically useful locations within the genome.
intrigue us here at WEEKLY. What is it about the science
CRISPR/Cas9 technology enables just that.
of CRISPR/Cas9 that is creating a surge of investor
euphoria? Let’s take a look at the science behind CRISPR stands for “clustered regularly interspaced
the controversy. short palindromic repeats”. These are brief DNA
sequences that read the same forward and backwards
BREAKING AND FIXING A GENE (palindromic repeats).
Genome-editing is possible thanks to damage in the In the mid-2000s, scientists realized palindromic
DNA sequence and the cell’s subsequent activation sequences were an integral part of the bacteria’s
of different repair pathways. These DNA breaks are immune system. Bacteria store away bits of invading
named double-stranded breaks (DSB) because both virus DNA between its own CRISPR (DNA) sequences
strands of the double-stranded DNA helix are broken. for memory purposes. If a virus eventually infects
Think of a two lane bridge that, after experiencing an the bacteria, the previously stored away viral DNA is
earthquake, has a section break off and fall into the copied and used to make viral RNA. When the viral RNA
water below. is released, it recognizes and binds to the DNA of the
Two kinds of repair pathways are charged with fixing subsequent invading virus. The binding action triggers
the break in the DNA: the protein Cas9 to cut up the viral DNA, knocking out
the virus.
• Non-Homologous End-Joining (NHEJ) closes the
gap between the break by joining the two sections In 2013, researchers showed they could use this same
back together—visualize pushing the two sides concept in animal cells. They simply designed sequence-
of the bridge together, leaving the fallen section specific RNA to escort Cas9 to a specific DNA site. Once
in the water. An unintended byproduct of NHEJ is at the site, Cas9 cuts the DNA sequence. This DSB
the possibility of sequence error, much like the results in disruption of the specific gene and activates
sections of the bridge not lining up properly; even either the NHEJ or the HDR repair pathway.

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EASILY CONFUSED: ZFN VS. CRISPR guide sequences and Cas9 protein into cells within
the patient’s body. Intellia Therapeutics licensed the
Zinc-finger nuclease (ZFN) is a protein engineered
technology from Caribou Biosciences (Berkeley, CA),
to bind and cut a specific genome sequence. CRISPR
which is also a contender in the CRISPR/Cas9 line up.
technology, on the other hand, relies on a RNA guide
sequence to identify and escort another protein Others are working on CRISPR/Cas9 delivery systems.
(Cas9) to the correct genome sequence. The Cas9 During fall 2014, Editas Medicine (Cambridge, MA)
protein is then responsible for the actual cutting of the published successful results on the use of adeno-
selected location. associated viral (AAV) gene therapy vectors to deliver
CRISPR/Cas9 to disrupt the expression of three target
CRISPR intended RNA sequences are quicker and less
genes in the neurons of mice. Editas recently licensed a
expensive to produce, as compared to engineering ZFN
Harvard-developed lipid-based delivery technology for
proteins. However, ZFNs have a longer track record and
CRISPR/Cas9.
are currently in the clinic being tested as a potential
cure for HIV/AIDS. Researchers are also developing CRISPR/Cas9
applications to fight infectious disease: Emory
THE CRISPR/CAS9 LINE UP University to combat hepatitis C virus and MIT to treat
fungal infections. Other early-stage academic research
Early, well-financed players in the CRISPR/cas9 arena
projects include targeting cystic fibrosis (Yale) and
include Intellia Therapeutics (Cambridge, MA), and
epigenome editing (Duke).
CRISPR Therapeutics (Basel, Switzerland). Both
announced plans to focus first on applications that With the race heating up, CRISPR/Cas9 is widely
involve modifying cells (like blood or bone marrow) expected to be utilized in a whole range of diseases,
outside of the body. The cells will be reinjected, including cancer, immune disorders, blood disorders,
bypassing (for now) the challenge of delivering the RNA rare genetic diseases, and even infectious diseases.

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THE RISE OF NANOMEDICINE Life Science Training from Industry Experts

BIOTECH TEAMS UP WITH NANOTECH NANOTHERM THERAPY

Imagine swallowing a pill containing microscopic MagForce’s (Berlin, Germany) NanoTherm is approved
magnets. After these “magnetic nanoparticles” make by the EMA to treat gliablastoma and sights are set
their way to the stomach, they are absorbed into the on a future FDA approval. NanoTherm is a highly
bloodstream to uncover disease associated proteins specific solid tumor treatment and is classified as
and other molecules. Envision these tiny explorers
a “ferrofluid”—a liquid that reacts to the presence
relaying their findings to a band worn discretely on
of a magnetic field. The water contained iron oxide
your wrist.
nanoparticles are the actual component of the therapy
The potential to identify disease early with that reacts to the magnetic field.
nanoparticles, rather than later with more conventional
technologies, is a game changer. In fact, Google X’s NanoTherm is injected directly into a tumor and the
(Mountain View, CA) life sciences arm announced this location subjected to a magnetic field. Iron oxide
very vision last fall. While the details of the plan are nanoparticles convert the magnetic energy into heat,
not public knowledge, their goal is to make this type of which is enough to either kill the tumor directly or to
nanotech available in five years. increase the tumor’s sensitivity to other treatments
Nanotechnology is defined as the understanding and such as chemotherapy. MagForce is conducting clinical
control of matter at an extremely small scale, and trials for prostate cancer in both the U.S. and E.U.
pairing it with biotechnology makes for some powerful
possibilities. In this WEEKLY we’ll take a look at a COCKTAIL FODDER: TINY DOCTORS
few applications a little less secretive than Google’s A nanotechnology lecture given in 1959, called
nanoscale disease detectives.
There’s Plenty of Room at the Bottom, may have very
well spurred the idea of nanomedicine. The idea of
3D NANOVACCINES
swallowing a “tiny doctor” to work on individual cells in
A 3D cancer vaccine released from nanomaterials the body certainly brightened the eyes of science fiction
is in the works at the Harvard Weiss Institute for writers at the time. Today, the roots of the field are
Biological Engineering (Cambridge, MA). It builds upon credited to the lecturer—Nobel Prize winning physicist
the concept behind Dendreon’s (Seattle, Washington)
Richard Feynman.
personalized prostate cancer immunotherapy Provenge.
3D vaccines are micro-sized silica rods suspended in NANOCOMBO: PARTICLES
liquid, injected under the skin, and once inside, the AND ANTIBODIES
rods reassemble into a three-dimensional structure.
The silica rods contain nano-sized pores loaded with Another magnetic nanoparticle approach to treating
a combination of chemical messengers and antigens cancer is in development at Bay Area biotech Accurexa.
meant to attract and activate dendritic cells. The freshly Instead of targeting solid tumors, their aim is to
activated dendritic cells diffuse out of the structure, “mop up” circulating tumor cells (CTCs). CTCs are
circulate throughout the body, and alert the immune cancer cells splintered off from a tumor that circulate
system when they encounter a hit (think cancer cell in the bloodstream and may lead to metastasis.
or pathogen). Antibodies (specific to the CTCs) attached to magnetic
3D vaccines can be quickly modified to target different nanoparticles are injected into a patient prior to
disease antigens by simply changing the load in the surgery. The concept: the antibody locks onto the CTC
nanopores of the silica rods—imagine that speed and the magnetic nanoparticle reacts to a magnet
and flexibility during epidemics and pandemics. and corrals the CTCs into one site. A magnetic field is
To date mice testing has shown significant delayed exacted on the congregated nanoparticles, where heat
tumor growth. destroys the CTCs. Plans are in the works for a clinical
trial in breast cancer patients undergoing surgery.

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TARGETING BREAST CANCER Life Science Training from Industry Experts

THE SUBTYPES OF BREAST CANCER also signals the patient’s immune system to attack
that cell.
Hearing your doctor utter the words HER2-positive,
HR-positive, triple-negative, or BRCA mutation can be Kadcyla, also made by Genentech, is an antibody-drug
devastating—even for the most resilient person. Simply conjugate—a monoclonal antibody that delivers a
put, all are linked to breast cancer. Breast cancer is highly toxic drug directly to HER2+ breast cancer cells.
complex and a diagnosis can be caused by all, some, or Kadcyla binds the HER2 receptor like Herceptin, but
even none of the factors listed above. also delivers a toxic payload (which is actually attached
to the monoclonal antibody). As a normal part of the
In fact, the National Cancer Institute’s (Bethesda,
cell’s lifecycle, cell-surface receptors get internalized or
MD) annual report to the nation recently outlined four
“taken up” by the cell on a regular basis. When Kadcyla
molecular subtypes of the disease. Each subtype is
is attached to a receptor that gets internalized, the
categorized by the cancer’s hormone receptor (HR)
toxic payload is released from the antibody and kills the
status and the level of expression from the HER2 gene.
cancer cell internally.
These cellular distinctions lead patients on different
treatment journeys because the cancer subtype HR-POSITIVE
determines the drugs used in a treatment plan.
About 70% of breast cancers diagnoses involve a
In this WEEKLY, we present a quick primer on the significant number of receptors for either estrogen
science behind HER2-positive, HR-positive, triple- or progesterone, making them hormone receptor
negative, and the BRCA gene. positive (HR+). HR+ cancers may respond positively
to treatments that block either the action or the
HER2-POSITIVE production of estrogen. In some cases, these
HER2-positive (HER2+) breast cancer patients—about treatments may continue to be used for up to five years
20% of all breast cancer cases—have the most highly after initial treatment in order to prevent recurrence.
effective therapies available on the market. HER2+
Two common types of medication for HR-positive breast
cancer cells produce, and therefore present, larger than
cancers are Tamoxifen and aromatase inhibitors. Both
normal numbers of the HER2 receptors on their cell
types of drugs may also be prescribed as a preventative
surface. These HER2 receptors capture growth factors,
treatment in women who are at high risk for breast
which trigger the cell to grow and reproduce more
cancer. In fact, Tamoxifen is named on the World
rapidly than normal. Mutations are more likely with
Health Organization’s List of Essential Medicines, a list
rapid reproduction and thus, a tumor is born.
of the most important medications needed in a basic
Overexpression of the HER2 receptor is the result healthcare system.
of having extra copies of the HER2 gene, known in
Tamoxifen works by inhibiting the estrogen receptor
the world of genomics as gene amplification. Gene
and was originally discovered by AstraZeneca (London,
amplification events are thought to be caused by
England). On the other hand, aromatase inhibitors block
mutations that occur after a person is born—it is not an
the production of estrogen by inhibiting an enzyme
inherited form of cancer.
whose activity is required for estrogen production. The
Genentech’s (South San Francisco, CA) Herceptin is different aromatase inhibitors on the market include
a monoclonal antibody that binds to and blocks the Arimidex (AstraZeneca), Femara (Novartis, Cambridge,
activity of the HER2 receptor on cancer cells. When the MA), and Aromasin (Pfizer, New York, NY).
HER2 receptor is blocked, the HER2 growth factor can
In February 2015, the FDA approved Pfizer’s Ibrance
no longer bind and send a growth signal to the cell,
for estrogen-receptor positive, HER2-negative breast
so the cancer cells stop dividing. The presence of an
cancer. Ibrance is a small molecule inhibitor of cellular
antibody on the surface of HER2+ breast cancer cells
enzymes known as cyclin-dependent kinases (CDKs).

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CDKs promote the development and division of cancer active in breast and ovarian cells, which is why certain
cells and inhibiting CDKs helps to arrest cancer growth. mutations in BRCA1/2 are associated with a significantly
increased risk of developing breast or ovarian cancer.
TRIPLE-NEGATIVE It must be stressed that BRCA1/2 mutations in and of
Triple-negative breast cancers lack receptors—they are themselves do not cause cancer; they simply make it
estrogen-receptor negative, progesterone-receptor more likely to occur.
negative, and HER2-negative. Since there are no A new class of drugs known as PARP1 inhibitors gives
receptor drug targets, this subtype is challenging to hope to women whose breast cancer is associated
treat and to date, there are no targeted therapeutics. with non-functioning BRCA genes. PARP1 is a second
If detected early enough, triple-negative breast cancer type of DNA repair protein. By inhibiting this pathway,
may respond well to chemotherapy. DNA damage becomes so extensive that the cancer
cells commit “cell suicide” (or apoptosis.) When the
THE BRCA GENE cell in question is a cancerous cell, apoptosis is a very
BRCA stands for “BReast CAncer susceptibility gene” good outcome.
and everyone has the BRCA 1 and BRCA 2 genes. The first FDA approved PARP1 inhibitor drug, Lynparza
The job of BRCA is to scan cellular DNA for damage (AstraZeneca) was approved for BRCA associated
and trigger DNA repair processes when mutations ovarian cancer in December 2014. AbbVie (North
are found. BRCA genes are passed down from one Chicago, IL) has a PARP1 inhibitor in Phase III for BRCA
generation to the next—a good thing, unless the version associated breast cancer.
passed down is a mutated variation.
Not all triple-negative breast cancers are BRCA
Mutated BRCA1/2 genes are non-functioning, so they associated, but many BRCA associated cancers are
cannot locate DNA damage, nor can they enlist DNA triple-negative. For this reason, triple-negative breast
repair. Testing positive for BRCA1/2 mutations may cancer patients may find hope in PARP1 inhibitor drugs.
indicate there is an accumulation of DNA damage,
which may eventually lead to cancer. BRCA is normally

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THE SCIENCE BEHIND THE BLOCKBUSTER
DRUGS OF TOMORROW Life Science Training from Industry Experts

BILLION DOLLAR MECHANISMS and thrive by evading T-cell detection. By inhibiting the
OF ACTION expression of PD-1 with Opdivo, the T-cell does not have
the means to read the signal from a PDL-1 protein and
The Most Promising Drugs of 2015—a Thomson
the T-cell attacks the tumor cells.
Reuters Cortellis Competitive Intelligence report—
includes several new drugs with predicted sales of $1 PRALUENT BY REGENERON
billion plus by 2019. Those on the list are well on their AND SANOFI
way to winning or have recently won FDA approval
•Drug type: Monoclonal antibody.
and the fervor around their potential has caught the
biotech world by storm. While stock gurus are passing •Drug class: PCSK9 inhibitor.
predictions on to investor circles, we here at WEEKLY •Indication: High cholesterol.
wonder: what exactly is the science propelling the
•Pipeline: FDA decision expected this summer.
market into overdrive? Let’s discover the mechanisms of
action driving the top three drugs on the list. Praluent works by binding to the PCKS9 protein, which
plays a key role in cholesterol metabolism. The PCKS9
OPDIVO BY BRISTOL-MYERS SQUIBB protein manages to trigger the degradation of low-
density lipoprotein (LDL) cholesterol receptors on the
• Drug type: Monoclonal antibody.
surface of liver cells. The function of a LDL cholesterol
• Drug class: PD-1 inhibitor. receptor is to “mop up” circulating LDL and their
• Indication: Approved by the FDA for melanoma and deterioration results in higher levels of LDL in the
lung cancer. patient’s blood. By preventing the degradation of these
Opdivo belongs to the new class of immunotherapy critical receptors, Praluent lowers LDL levels to lessen
drugs known for their inhibition of the PD-1 protein the risk of a cardiovascular event.
found on T-cells. Under normal conditions, the PD-1
protein is used by the immune system to prevent an
LCZ696 BY NOVARTIS
attack on the body it is charged with protecting. When •Drug type: Small molecule drug combination.
PD-1 encounters a cell with PDL-1 proteins, it identifies •Drug class: Angiotensin-neprilysin inhibitor.
the cell as “self” and signals the T-cell to hold back and
•Indication: Heart failure.
not attack.
•Pipeline: FDA decision expected in August.
Some cancers have a sneaky tendency to overexpress
PDL-1 in their cells and this helps tumors to survive LCZ696 is a combination of the drugs Valsartan and
Sacubitril. This dynamic duo work together to lower
blood pressure, effectively reducing the strain on the
heart and lessening fluid accumulation in the tissues
(such as the lungs)—all key symptoms of heart failure.
Valsartan is an angiotensin II receptor inhibitor—it
stops angiotensin II. When angiotensin II attaches to the
angiotensin receptor, blood vessels constrict and blood
pressure rises. Think about it like this: it takes more
work to push fluid through a long narrow tube than
through a wide one.
Angiotensin II also promotes the release of a second
hormone, aldosterone, which increases sodium
retention by the kidneys and further drives up blood

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pressure. Valsartan blocks the angiotensin receptor EASILY CONFUSED: AGONIST
so angiotensin II has nowhere to land, resulting in no VS. ANTAGONIST
reaction cascade, no blood vessels constriction and
An agonist is a chemical that binds to a receptor and
ultimately blood pressure is lowered.
activates it to produce a biological response. An
Sacubitril is a neprilysin inhibitor—it stops neprilysin. antagonist blocks the action of an agonist—in other
Neprilysin naturally breaks down natriuretic peptide, words, it acts as an inhibitor. The three drugs featured
a hormone whose job is to get rid of sodium and in this WEEKLY, as well as with a majority of drugs
dilate blood vessels. By inhibiting neprilysin, Sacubitril currently on the market, are antagonists.
allows the hormone to reign free and blood pressure
Want to bend your mind a bit further? An inverse
is lowered.
agonist causes a biological response opposite to that of
By combining an angiotensin II receptor inhibitor and the agonist.
a neprilysin inhibitor, LCZ696 drives drown blood
pressure and decreases the rate of heart failure.

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GETTING RID OF ALLERGIES Life Science Training from Industry Experts

TAKING A SWING AT ALLERGENS allergic reaction has occurred, however, it is too late for
antihistamines to be effective.
Watching a game at the ballpark and digging into a
bag of peanuts is a source of entertainment for many Histamines are chemical messengers that trigger a
Americans. For the 15 million who sufferer from peanut whole spectrum of different symptoms ranging from
allergies, the idea of being taken out to the ballgame annoying to deadly, including:
elicits concern—or even anxiety. • Increase in blood vessel permeability that allows
Food allergies—think tree nuts, milk, eggs, wheat, fluid to escape, resulting in a runny nose and
soy, fish, and shellfish—are on the rise. The mere dust watery eyes.
particle of a freshly cracked peanut can be responsible • Increase in smooth muscle contraction that leads to
for an unpredictable cascade of reactions, including throat constriction and difficulty breathing.
death brought about by anaphylaxis. • Extreme tissue fluid release that causes a sudden
This WEEKLY takes a swing at explaining how allergies drop in blood pressure, potentially bringing on a
develop, are currently treated, and what new products heart attack.
might change the way allergen desensitization therapy • Difficulty breathing and swallowing, swelling, heart
is delivered. palpitations, and unconsciousness—occasionally
causing death.
SOMETHING TO SNEEZE AT
The latter symptoms are known as anaphylaxis.
The host of symptoms dubbed “allergies” are the
Anaphylaxis is treated with an injection of
end result of the immune system responding to a
epinephrine—and the sooner, the better in cases
normally harmless substance, as if it were a threat. An
where a life is on the line. Epinephrine helps to reverse
initial allergen exposure results in the production of
histamine’s effects by stimulating the reformation
a particular class of antibodies called IgE antibodies.
of intact blood vessels, promoting the relaxation of
Allergies develop if excessive amounts of IgE antibodies
smooth muscle cells, and stimulating the heart. People
are produced.
at risk for anaphylaxis need access to an epinephrine
IgE antibodies bind to a specialized type of white blood autoinjector—a spring-loaded syringe that makes the
cell called a mast cell. When exposed to the allergen lifesaving injection readily available. The best known
a second time, multiple allergen-IgE complexes bind epinephrine autoinjector is Mylan’s (Canonsburg, PA)
to mast cells, triggering a release of histamines in an EpiPen. Other combination medical devices on the
attempt to get rid of the allergen. Only some of the IgE market include Amedra’s (Horsham, PA) Adrenaclick
antibodies need to recognize the allergen as harmful in and Sanofi’s (Paris, France) Auvi-Q.
order for the histamine release to occur.
Histamine binds to receptor proteins on the surface REVISITING THE HYGIENE HYPOTHESIS
of blood vessels, which results in the modification of Epidemiologists have noticed an interesting trend as
cadherin proteins. Cadherin proteins are intercellular countries rise from developing to developed status:
proteins with the job of helping cells to stick together. an improvement in sanitation and access to antibiotics
Gaps then form between the cells that make up the means less pathogenic exposure and lower infection
blood vessels as a result of the modification, allowing rate. As the infection rate drops, the incidence for
fluid to escape. Histamines may also bind to receptors different kinds of allergies shoots up.
on certain types of nerve cells, resulting in smooth Many scientists think early exposure to infection helps
muscle contraction—and in some cases a sensation shift the immune response towards fighting pathogens
of itchiness. Antihistamines work by blocking the while minimizing the production of IgE antibodies.
activation of the histamine receptor. Once a severe Exposure to potential allergens (while the immune

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system is still developing) helps to desensitize the monitoring by a physician) and drops or tablets
allergic response. dissolved under the tongue and swallowed (which can
sometimes be taken at home). Allergen Research
THE STRATEGY BEHIND Corporation’s (San Mateo, CA) AR 101 is made of
DESENSITIZATION THERAPY pharmaceutical grade peanut proteins and can be
The idea of allergy desensitization through controlled mixed with food as a means of delivery. Positive
exposure has been around for decades. Desensitization Phase II results prompted the FDA to award Fast Track
is the principle behind allergy shots proven to be designation—and the promise of AR 101 reeled in $80
effective against pet dander, dust mites, and pollen. million in funding earlier this month.
Desensitization therapy was once considered to be AR 101 Phase II studies report patients becoming
too risky for food allergies, but a number of new desensitized to doses at least twenty times greater
studies support the idea that gradual exposure to food than the original allergy inducing dose—and in some
allergens may also be beneficial. cases, more than 100-fold greater. The goal is to reach a
With this strategy in mind, the Immune Tolerance tolerance level that offers protection against accidental
Network (Seattle, WA) recently announced the results ingestion of peanuts, potentially avoiding a deadly
of a study that dialed the idea of desensitization reaction. As Allergen Research Corporation heads
back even further. Their hypothesis was that early towards Phase III for peanuts, plans for clinical studies
exposure to potential allergens can actually prevent the on immune system training for egg and milk allergies
development of allergies. Six hundred forty children will begin later this year.
between four and eleven months old (all of whom were DBV Technologies (Bagneux, France) is also in the fight
considered to be at risk for developing allergies) were against allergies. They are currently conducting clinical
divided into two groups: peanut avoider or peanut trials on their Viaskin skin patch that delivers low doses
consumer. The avoiders were given no peanut products of either peanut proteins, milk, or dust mite allergens.
until age five, while the consumers were given several By delivering an allergen through the skin instead of the
snacks containing peanuts each week. Astonishingly, blood, the body will not react as severely, lessening the
by the time the children reached age five, 17% of the risk of anaphylaxis as a side effect. The Viaskin peanut
avoiders had developed peanut allergy compared to patch—which is set to enter Phase III by year’s end—has
only 3% of the peanut consumers. been awarded Fast Track designation by the FDA. As
allergen desensitization treatments continue to make
ALLERGENS BY THE DOSE their way through the drug pipeline, allergy sufferers
The current allergen immunotherapy market includes remain hopeful for better and easier treatment options,
the above mentioned allergy shots (which require and maybe even one day—a cure.

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THREE PARENT BABIES APPROVED Life Science Training from Industry Experts

THREE PARENT IN VITRO procedure was used to help woman, whose fertility had
FERTILIZATION COMING SOON declined, to conceive.

The U.K. Parliament green-lit “Three Parent In Vitro The prospective mother’s egg was injected with a
Fertilization” (TPIVF) legislation last month, setting a small amount of cytoplasm from another women’s egg
precedent and spawning headlines around the globe. prior to fertilization. Cytoplasm is the liquid portion
Dubbed “three parent babies” by news outlets, the of a cell outside of the nucleus that also happens
law will allow the procedure to move directly into to contain mitochondria. The exact mechanism by
fertility clinics. which cytoplasmic transfer enables pregnancy is not
clearly defined, but many doctors suspect some cases
The process combines donor mitochondrial DNA with of infertility are caused by damaged mitochondria.
in vitro fertilization to circumvent mitochondrial-based The infusion of donor cytoplasm may be enough to
diseases in offspring. Defects in mitochondrial function rejuvenate these eggs.
are associated with various musculoskeletal, metabolic,
and neurodegenerative diseases. Twenty-four women achieved pregnancy via
cytoplasmic transfer two decades ago. However,
In this issue we will explain the science behind TPIVF the FDA had safety concerns and the practice was
and look at its potential for approval in the U.S. abandoned when fertility clinics were required to file an
IND to continue the procedure. Cytoplasmic transfer is
REPLACING MITOCHONDRIAL still not approved by the FDA and this policy is credited
DNA IN IVF with creating a market for reproductive tourism abroad.
The rationale behind TPIVF lies with mitochondria—
what many of us remember as the “powerhouse” of COCKTAIL FODDER: ONLY 37 GENES
a cell from high school biology. Recall they are the Although it is correct to say that a baby conceived by
compartments that convert glucose into the energy TPIVF has genetic material from three different parents,
our cells use to do work. What you may not remember the vast majority (~99.9%) of that baby’s genetic
is mitochondria have their own DNA that is inherited material will come from the nuclear DNA of the original
maternally. When an egg is fertilized, the mitochondria egg and sperm. Mitochondrial DNA codes for only 37
get passed on in subsequent rounds of cell division genes, whereas nuclear DNA codes for ~21,000 genes.
becoming a part of every cell in a developing baby’s
body. A woman with defective mitochondrial DNA THE FDA WEIGHS IN
passes this trait onto her child. However, these
mutations may be weeded out of the genetic landscape Much like IVF involving donor eggs, TPIVF has the
thanks to TPIVF. potential to become a routine medical procedure, but
not without some close scrutiny. The FDA has asked the
During TPIVF the nuclear DNA is removed from the egg Institute of Medicine (Washington, DC) to consider
of the afflicted mother. This nuclear DNA is transferred the ethical and social ramifications raised by TPIVF
into an enucleated donor egg (enucleated means the and to create a consensus report to help guide future
nuclear DNA has been removed but the mitochondrial regulatory policy. As a part of this process, the Institute
DNA remains intact). The resulting egg is implanted of Medicine will hold a series of meetings and solicit
back into the prospective mother and (fingers crossed) public commentary. The first meeting was held in
develops into a healthy baby. The resulting child has January and the second meeting will be held March 31–
DNA from two different women and one man. April 1 in Washington D.C.

BEGINNING WITH Click here to register for the public comment portion,
CYTOPLASMIC TRANSFER registration ends tomorrow, March 27, at 2pm.

TPIVF has some precedent in a procedure known as

cytoplasmic transfer. In the late 1990s this experimental

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THE POWER OF ONCOLYTIC VIRUSES Life Science Training from Industry Experts

HBO SHINES LIGHT ON Oncolytic viruses have yet to be approved by the FDA,
NOVEL IMMUNOTHERAPIES but several are in clinical trials. Imanis Life Sciences’
(Rochester, MN) MV-NIS is an oncolytic measles virus
HBO’s Vice news program selected a few promising
in Phase I/II. DNAtrix’s (Houston, TX) DNX-2401 is an
technologies from the immunotherapy pipeline to
oncolytic adenovirus that just completed Phase I trials.
feature in its Killing Cancer episode. Vice’s heartrending
Both were featured in the Vice episode.
human interest stories illustrated the potential of three
different developmental drugs: The closest to market entry is Amgen’s (Thousand
Oaks, CA) Talimogene laherparepvec (T-Vec), a modified
• An oncolytic measles virus battling myeloma.
herpes simplex 1 virus. T-Vec is engineered to target
• An oncolytic adenovirus fighting a brain tumor. melanoma and express GMCSF to fight cancer cells
• An engineered human immunodeficiency virus (and not cause cold sores). With Phase III complete, an
knocking out childhood leukemia. FDA decision is expected in the near future. Amgen is
also aiming to expand the T-Vec market with an EMA
While this program has generated buzz around the
submission in process.
hope for a cure, we were left to ponder: what exactly
is the science behind the scenes? Let’s explore how
INSIDE CAR-T THERAPY
oncolytic viruses battle cancer.
Scientists are engineering T-cells to recognize a specific
TERM OF THE WEEK: IMMUNOTHERAPY tumor antigen, as seen in the case of young leukemia
survivor Emily Whitehead.
Immunotherapy is a type of therapy that harnesses
the power of a patient’s immune system to combat The human immunodeficiency virus is engineered using
a disease. the following process:
• T-cells are removed from the patient’s blood.
ONCOLYTIC VIRUSES EXPLAINED • HIV is prepared by researchers that strip their
An oncolytic virus is a virus that infects cancer cells ability to cause illness; however, they retain their
and multiplies inside of them, potentially rupturing and capacity to integrate into cells’ DNA. These viruses
killing the individual cancer cells. are then used to deliver genetic material to the
Oncolytic viruses are created in the lab by genetically patient’s T-cells in the lab.
modifying existing viruses in at least two ways: • This results in a modified T-cell receptor, called
• Making the virus safe by removing genes that cause a chimeric antigen receptor T-cell (CAR-T), that is
the virus to make people sick. able to recognize proteins on the surface of the
patient’s tumor.
• Engineering viral surface proteins so the virus
recognizes and binds to the cell receptors of • The newly formed CAR-T cells are then multiplied in
cancerous cells, disregarding the healthy, non- the lab.
cancerous cells. • This group of CAR-T cells are infused back into the
Additional modifications can be made to further patient’s blood, where they ferret out and destroy
equip an oncolytic virus, such as adding an immune tumor cells.
stimulating gene to produce a granulocyte-macrophage CAR-T therapy originated from research done by Dr. Carl
colony-stimulating factor (GMCSF). GMCSF works to June at the University of Pennsylvania (Philadelphia,
activate the patient’s own immune system and target PA), and was subsequently licensed and developed by
the tumor. Novartis. No CAR-T therapy has been approved by the
FDA, but Novartis’ CTL019—used on Emily Whitehead—

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is in Phase II. The CAR-T market is swelling with other In 1904, a paper published in American Journal of
players such as Amgen, Kite (Santa Monica, California), Medical Science described the spontaneous regression
Juno (Seattle, WA), Celgene (Summit, NJ) and Cellectis of cervical cancer following a rabies vaccination. A
(Paris, France). few years later, this observation was followed by an
If engineered T-cells continue to live up to the hype, account of the remission of lymphomas after measles
we can look forward to what some have termed a virus infection.
“living drug”—cancer fighting cells that will persist and The recent ability to engineer viruses to remove genes
multiply in a patient’s body until the tumor is destroyed. that create illness combined with the addition of
specific tumor targeting genes make this century old
COCKTAIL FODDER: NEW speculation a likely reality.
TECHNOLOGY, OLD OBSERVATION
While the idea of using viruses as a vector to fight
cancer are gathering steam in the mainstream press,
the technology has been around for over 100 years.

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INTERCHANGABILITY DENIED TO
FIRST US APPROVED BIOSIMILAR Life Science Training from Industry Experts

G-CSF: THE ORIGINAL INNOVATOR affect the final structure. Cell lines originating
from different manufacturers are not identical,
Sandoz’s (Princeton, NJ) Zarzio rode the first wave
even if both are Chinese hamster ovary cells—a
into the U.S. biosimilars market after it received
biomanufacturing favorite.
FDA approval last week. Zarzio is similar to Amgen’s
(Thousand Oaks, CA) Neupogen and both of these These factors give rise to the saying “the product is the
medications are recombinant versions of the “go to” process,” and is the reason biosimilar drugs are not
stimulator for white blood cell production—known as considered identical to the source biologic. Therefore,
granulocyte-colony stimulating factor (G-CSF). biosimilars are required to undergo at least some
clinical testing to confirm safety and efficacy prior
White blood cells (WBC) are the front line troops that
to approval.
protect the body from infection and disease and are
produced when G-CSF is released from bone marrow. TERM OF THE WEEK:
G-CSF beefs up the immune system when it binds WHITE BLOOD CELL
to receptors on the precursor to WBCs, known as
White blood cells, also called leukocytes, are any of
hematopoietic stem cells. This attachment initiates the
the various colorless cells of the immune system
differentiation and maturation of the stem cell so it can
that circulate mainly in the blood and lymph. B-cells,
graduate to become a WBC.
T-cells, macrophages, monocytes, and granulocytes are
Patients undergoing chemotherapy and bone marrow specialized white blood cells.
transplants run low on WBCs and an infusion of
Neupogen or Zarzio elicits the same G-CSF action— INTERCHANGEABLITY DENIED
increased immune response. While both drugs are
Zarzio and Neupogen are approved for the exact same
potential lifesavers, the real story is how the FDA
indication. The twist is that Zarzio is not designated
decided to classify these two biologics. Let’s take a look
as interchangeable with Neupogen. Therefore, a
at the science and policy behind biosimilars.
pharmacist is not allowed to substitute one drug for
another because the FDA does not consider Zarzio to be
SIMILAR, NOT IDENTICAL
identical to Neupogen. Here is a fact highlighted in their
Biologic drugs are made by transferring the gene for different International Nonproprietary Names (INN):
a therapeutic protein into a bacterial or mammalian Neupogen’s is filgrastim and Zarzio’s is filgrastim-sndz.
cell. That production cell makes the protein that
Why is the FDA hesitant to give a biosimilar
corresponds to the transferred gene.
interchangeable status with the brand name biologic?
Gene sequences for therapeutic proteins are publicly Immunogenicity—the ability to provoke an immune
available. In theory it should be straightforward to response. All biologics are potentially immunogenic, so
make a copycat version of any biologic drug. In practice, switching biologics is a safty concern.
however, many external factors influence the final,
intricately-folded protein structure. Recall that it is THE CHASE IS ON
imperative the protein be folded correctly because
Biosimilars stem from Congress’ passage of the Patient
protein structure influences protein function. If the
Protection and Affordable Care Act five years ago. The bill
protein structure is not correct, the protein may
authorized the FDA to approve biosimilars as a means
not work.
to lower the cost of biologic drugs, which Congress
The precise manufacturing conditions are considered hoped would herald in a new era for healthcare. Are
proprietary because the process of growing cells lower healthcare costs for biologics eminent? Below
influences the final structure of the protein. Even are some top selling biologics and the biosimilar rivals
slight differences in the cells themselves can chasing them.

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 Learn more about biosimilars. Check out our earlier
issue, Biosimilars: Ready Or Not, Here They Come.

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IT’S A GREAT TIME TO BE IN BIOTECH Life Science Training from Industry Experts

It’s a great time to be in biotech—new therapeutics are Genome-editing therapy engineers double-stranded
breaking ground faster than we can say “monoclonal breaks to occur at specific locations, activating the
antibody” and the technologies behind them are heavy intrinsic cell repair pathways: HDR and NHEJ.
with promise and potential. Sangamo Therapeutics is applying zinc finger nuclease
Our mission is to keep your industry knowledge up (ZFN) genome-editing techniques to disrupt the CCR5
to date and the WEEKLY is here to give you a smart gene of HIV patients’ T-cells—this stops the virus from
primer on the basics. Below we have outlined several infecting new cells and restores the immune system.
of the therapeutics and innovators making waves in The first clinical trials for genome editing are underway
the industry. for Sangamo’s SB-728, the therapy is currently in Phase
II testing .
GENOME-EDITING: IT’S ALL
ABOUT THE BREAK
Genome-editing is the ability to selectively disable or
edit the sequence of specific genes. This genome-based
therapy is made possible thanks to the cell’s natural
ability to repair DNA damage. DNA damage occurs as
double-stranded breaks (DSB) because both strands of
the double-stranded DNA helix are broken, similar to
a two lane bridge that has a section break and fall off
after experiencing an earthquake.
There are two types of DNA repair pathways the cell
uses to fix DSBs: Non-Homologous End-Joining and
Homology Directed Repair.
• Non-Homologous End-Joining (NHEJ) closes the
Zinc finger nuclease genome-editing explained.
gap between the break by joining the two sections
back together—imagine pushing the two sides of To read more, check out Designer Genes: An
the bridge together, without including the section Introduction to Genome-Editing.
that broke off. An unintended byproduct of NHEJ
is the possibility of sequence error, much like the ANTIBODY-DRUG CONJUGATE:
sections of the bridge not lining up properly; even IT’S ALL ABOUT THE DELIVERY
a single base deletion may cause unintended
Monoclonal antibodies (mAb) are now being used as
consequences. If the repair occurs in the middle of
couriers to deliver a toxic drug to target cells—known as
a gene, the minor error can be enough to disrupt
antibody-drug conjugates. In this application, a highly
gene function and halt the production of the
toxic compound is chemically attached to a mAb that
corresponding protein.
recognizes proteins on the surface of a cancer cell. Once
• Homology Directed Repair (HDR) relies on a highly bound, the deadly payload is internalized and delivered
similar (homologous) DNA segment to repair the to the innards of the tumor cells. This therapy is less
break—imagine the missing bridge section built toxic because it is only delivered to the cancer cells,
elsewhere and helicoptered in to fill the break. leaving the neighboring healthy cells in the patient’s
body relatively unharmed.

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 The above image represents the two arms of a bispecific antibody.
The toxic “drug bomb” attached to the mAb above
successfully annihilates the target cell. To read more, check out The Bispecific Antibody: A
Lethal Hybrid.
While several antibody-drug conjugates are in the midst
of development, only two antibody-drug conjugates STEM CELL THERAPY: IT’S
have made it to market. Roche’s Kadcyla is taking out ALL ABOUT THE MIX
breast cancer and Seattle Genetics’ Adcetris mutes
Stem cell therapy introduces new cells into tissue to
cancerous lymphocytes in lymphoma.
treat disease. The scientific challenge is to figure out
To read more, check out Biotech’s Battlefront: the exact cocktail of growth factors, hormones, and
Monoclonal Antibodies. nutrients required to lead a stem cell down the correct
and intended developmental path. Once the cocktail
BISPECIFIC ANTIBODY: ITS is perfected, scientists can differentiate cell types
ALL ABOUT THE HYBRID into their choosing. The ultimate goal is to use these
A bispecific antibody is a genetically engineered protein new cells to produce replacement tissue and organs
composed of two different monoclonal antibody for patients suffering from degenerative diseases or
fragments, where one fragment binds to the target cell traumatic injuries.
and the other fragment to a killing agent. The killing Embryonic stem cells are prized because of their
agent is a specialized T-cell, called a Killer T-cell. This potential to develop into any human tissue—a
T-cell naturally makes up the front line of the specific characteristic called pluripotency. By reactivating the
immune response. Once activated, Killer T-cells can four genes turned off during the progression from
be highly effective cancer killing machines. However, embryonic stem cell to specific tissue type, researchers
they often miss tumor cells because tumor cells are can turn back the hands of time and create induced
not recognized as “foreign”. So the drug discovery pluripotent stem cells (IPSC). The IPSC advantage means
challenge has been figuring out how to create an less chance of rejection by a patient’s body.
army of T-cells that recognize tumor cells—enter the
To date, the only stem cell-based therapy approved
bispecific antibody.
by the FDA is New York Blood Center’s Hemacord,
Blincyto, Amgen’s acute lymphoblastic leukemia a cord blood product indicated for disorders related
treatment, redirects cytotoxic T-cells to target specific to production of blood in the body. Clinical trials are
tumor cells. Blincyto coasted past the FDA bottleneck ongoing for stem cell derived therapies for diabetes,
ahead of schedule, and gained approval in December stroke, ALS, and spinal cord injury.
2014 as a first-of-its-kind immunotherapy.
To read more, check out Tailoring Stem Cells to Fashion
Replacement Organs.

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NEW BRAIN CELLS FROM A PILL Life Science Training from Industry Experts

Imagine being able to regenerate your brain cells by Chemical libraries may be customized for a particular
swallowing a pill. The potential treatment relies on a company’s interests—for example, having structures
process known as neurogenesis—”neuro” meaning likely to inhibit certain types of signaling pathways are
nerves and “genesis” meaning creation. Adults suffering of interest to companies focusing on oncology drug
from neurological diseases or brain disorders may one development. In the case of scientists developing
day benefit from small molecule activators to promote neurogenesis compounds, the ability to cross the blood-
the birth of neurons in the brain. brain barrier is paramount, because the protective
Neurogenesis naturally occurs during fetal brain network of blood vessels envelopes the brain and
development and declines with age. Adult neurogenesis prevents the entry of most substances.
was assumed to be nonexistent until twenty years ago,
when it was discovered that the hippocampus could in
MICE AS A TESTING DEVICE
deed regenerate neurons. Mouse models clue us in as to how a particular
drug might work in a human patient and are more
A decrease in hippocampus neurons is associated with
comprehensive than testing using lab grown cells.
Parkinson’s and Alzheimer’s disease, and may be the
cause of memory loss and disorientation commonly Genetic engineering is used to alter a mouse to have
associated with both conditions. Recent studies also genes that mimic genetically based human diseases.
suggest chronic stress slows neurogenesis, indicating a In the case of testing drugs for cystic fibrosis, one
possible association with clinical depression. of the mutations associated with the disease would
be genetically engineered into the mouse and then
In this WEEKLY we’ll learn the nuances of drug
tested accordingly.
discovery to get a realtime assessment of the current
neurogenesis line up. Since genetic associations are not as clear cut for most
diseases, mice used to study Alzheimer’s, for example,
CHECKING OUT THE are elderly mice whose reduced hippocampal volume
CHEMICAL LIBRARY and cognitive decline mirror those of elderly human
In drug discovery we often hear about companies using Alzheimer’s patients. A mouse model of Parkinson’s
a chemical library—a collection of different chemical disease is created by treating mice with a chemical
compounds, typically on the order of hundreds of called MPTP that destroys dopaminergic neurons—a
thousands of different compounds. Each compound has hallmark of the disease.
associated information, such as chemical structure and
characteristics, catalogued in a database.
TERM OF THE WEEK: NEUROGENESIS
Neurogenesis is the process by which neurons are
Each compound in the library is screened or tested
generated from neural stem cells and progenitor cells.
for its ability to have a particular effect in a cell-based
Adult neurogenesis is the process of generating new
assay (test). For example, different small molecule
neurons which integrate into existing circuits after fetal
compounds could be screened for their ability to
and early postnatal development has ceased.
activate neurogenesis from neuronal stem cells grown
in the lab. If a compound shows promise, it is called
a “hit”; it is then modified further and its derivatives
IN THE CLINIC
are tested to see if they show even greater efficacy. Neuralstem (Germantown, MD) and Neuronascent
After several rounds of modification, scientists will (Clarksville, MD) are leading the charge in developing
present a lead candidate to advance into preclinical small molecule activators of neurogenesis. By
animal testing. screening large chemical libraries, they have identified
various compounds showing promise in their ability
to activate neurogenesis from adult neural stem

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cells, both in the lab and in mouse models of various Neuronascent is focusing on Alzheimer’s in their
neurodegenerative disorders. approach to neurogenesis drug development. In a
Neuralstem’s lead neurogenesis candidate, NSI-189, mouse model of Alzheimer’s, lead compound NNI-362
increased the hippocampal region of mouse brains by promoted the growth of new hippocampal neurons that
as much as 20%. With Phase I trials for major depressive not only migrated to the correct functional location but
order recently completed, plans to enter Phase II clinical also differentiated and survived long enough to reverse
testing are expected by mid-2015. previously observed cognitive declines. Neuronascent is
preparing for Phase I trials of NNI-362.

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GMO A DAY KEEPS DOC AWAY?
THANKS TO RNAI APPLES Life Science Training from Industry Experts

The latest apple out of the orchard, the Arctic apple, gene to the ribosome to make protein. RNAi technology
just so happens to resist the ugly browning kids reject creates a double-stranded RNA (dsRNA) by introducing
if sliced. So, how does biotech factor into the equation? an RNA sequence into a cell which is complementary
Genetic engineering. to the RNA for the particular gene to be silenced. When
This first-of-its-kind apple, approved by the USDA both ssRNAs meet, dsRNA is formed. The cell recognizes
last week, caught our eye here at WEEKLY because it dsRNA as “foreign” and destroys it.
represents a new type of genetically modified food— Short segments of apple DNA, coding for the four
one that has been engineered to directly appeal to and different PPO genes, were transferred to make the
benefit the consumer, rather than the farmer. Because Arctic apple. Once expressed, the short segments of
of the newer technology used in the creation of the DNA produce a segment of RNA complementary to
fruit, lingering safety concerns voiced by critics may the PPO RNA already being made by the apple. The
begin to fade. resulting dsRNA is immediately destroyed by the
Let’s take a look at how biotechnology silences the cell. No PPO equates to Arctic apples’ “just sliced”
apple browning gene and visit its application in the field appearance.
of genetically modified food.
IN THE INDUSTRY
WHY SO BROWN? All facets of the industry, such as the growers, packers,
The browning observed in conventional apples is shippers, and retailers will benefit from the stoppage
the end result of an oxidation reaction. The reaction of the oxidation process. Food processors, likewise, will
is first catalyzed by an enzyme called “polyphenol be able to produce more consistent juices, sauces, and
oxidase” (PPO), which acts on a class of compounds sliced apple products without relying on antioxidant
called phenolic compounds, present in various fruits. In treatments currently in use.
order for the reaction to occur, PPO has to come into The technology also allows for better discernment
contact with its substrate—the phenolic compounds. between simple bruising and a rotten apple. With
These compounds are typically sequestered inside of oxidation no longer an issue, brown discolorations on
vacuoles, or internal compartments of a cell, which are an apple will more likely indicate rot, meaning more
enclosed by a membrane and filled with water. sellable product and less waste.
If the cell is disrupted by slicing or dropping, the
vacuoles burst and allow the phenolic compounds
TO THE MARKET
to come into contact with PPO, resulting in the Since Arctic apples contain no foreign DNA (the 4
brown tinge. introduced genes are apple DNA), they are likely to
impart a truer flavor to GMO wary consumers. Having
TURN THE BROWN UPSIDE DOWN undergone ten years of field testing, Arctic apples
Okanagan Specialty Fruits (Summerland, B.C., do not differ in any significant respect to unmodified
Canada), the maker of the Arctic apple, sought to halt apples, apart from the lack of PPO enzyme.
the oxidation of phenolic compounds. They chose to The USDA approval grants permission to market
tackle the problem by blocking the production of the PPO the plants to growers, so the first two varieties of
protein; no enzyme, no chemical reaction, no brown. modified apples, Golden Delicious and Granny Smith,
How? By using a gene silencing technique known will be available in Fall of 2016. If the Arctic apples are
as RNAi. successful, they could pave the way for other consumer
Recall from high school biology that RNA is single- oriented products, such as oxidation-resistant cherries
stranded (ssRNA) and is the set of instructions from the and pears, which are in development at Okanagan
Specialty Fruits.

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A similar product, the Innate Potato, was approved last the same technology. When potatoes are cooked at high
November by the USDA. Developed by J.R. Simplot temperatures, asparagine synthetase-1 reacts with the
(Boise, Idaho), the spud also uses RNAi technology to potato sugars to produce a chemical called acrylamide
decrease production of the PPO protein in order to which has been linked to cancer in rodents. Simplot
reduce browning. plans to launch the potato in limited test markets during
The production of a second protein in Innate potatoes, spring of 2015.
asparagine synthetase-1, is also knocked down using

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CURING HEPATITIS C Life Science Training from Industry Experts

The biotech industry gets its fair share of criticism. Take The high mutation rate and the challenge of targeting
for example HBO’s John Oliver shining his light on a virus without negatively impacting the host cells are
biopharma last week and proclaiming drug companies major hurdles for antiviral drug development. So it is a
are like high school boyfriends: more interested BIG deal when a truly safe and effective antiviral drug
in getting into patients’ bodies than actually being makes it to the marketplace.
effective once they’re inside.
That soundbite prompted us at the WEEKLY to wonder:
WHAT IS HEPATITIS C?
Is a 90% cure rate for a disease afflicting 3.5 million in Hepatitis is a liver disease caused by infection with
the U.S. and possibly 200 million worldwide effective? the hepatitis C virus (HCV), a blood borne pathogen.
Chronic infections may initially be asymptomatic, and
We’re talking about hepatitis C and the new generation
if left untreated, serious problems such as cirrhosis
of drugs put on the market just last year to cure it:
(scarring) or cancer of the liver can occur. Liver disease
Gilead’s (Foster City, CA) Harvoni and Sovaldi, and
due to HCV infection is the leading indication for liver
AbbVie’s (North Chicago, IL) Viekira Pak.
transplants in the United States. Currently there is no
Before these drugs, hepatitis C patients could expect vaccine available for HCV.
a standard treatment involving a combination of
interferon and ribavirin. Interferon is delivered via AN EASIER PILL TO SWALLOW:
repeat injections to patients and works by “revving up” SOVALDI AND HARVONI
the entire immune system, which leads to unpleasant
Small molecule drug Sovaldi, approved at the end of
side effects such as fatigue, anxiety, flu-like symptoms
2013, has been widely heralded as the beginning of a
and gastrointestinal distress. In exchange for those
new era in HCV treatment. Marketed by Gilead, Sovaldi
unpleasant side effects, that drug combo has a ~50%
was granted breakthrough therapy status by the FDA
success rate.
because it is more effective and has fewer side effects
Contrast with Harvoni’s ~90% hepatitis C cure rate, than interferon with ribavirin.
and we can all agree this therapeutic is a bright spot
Sovaldi can be used in combination with interferon
for our industry and a story that deserves to be told.
and ribavirin, or ribavirin alone, depending on when
Let’s break down the science behind the new drugs
the HCV is diagnosed and the severity of the disease.
fighting hepatitis.
Sovaldi conquers HCV from a bottom-up approach,
disrupting the viral replication stage. Interferon fights
SIMPLE VIRUS, COMPLEX PROBLEM
HCV from the top-down, prompting the entire immune
For all the damage some viruses can cause, they’re system to beef up an assault.
actually very simple structures. Consisting of genetic
As a nucleotide analog polymerase inhibitor, Sovaldi
material (either RNA or DNA) and a batch of proteins
inhibits the viral polymerase –the enzyme used by HCV
to aid with host cell infection, viruses stealthy infiltrate
to replicate its genetic material (RNA). To make copies
our bodies.
of the viral RNA, the polymerase simply connects new
Though viruses cannot make their own proteins, they building blocks (nucleotides) together in the same
are generally able to make copies of their own genetic order as the existing viral RNA. The analog drug is
material by using the polymerase proteins found inside structurally very similar to nucleotides found in nature,
the infectious particle. Most RNA-based viruses, such as so the polymerase will subsequently incorporate it into
the HIV, HCV and influenza, have polymerase enzymes a growing RNA strand. However, it has been chemically
with a very high error rate—in other words, they make a modified so once incorporated, the polymerase is
lot of mistakes when copying the viral genome, resulting unable to add any additional nucleotides, thereby
in mutations. halting viral replication.

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In October 2014, the FDA approved Harvoni, a
combination product consisting of Sovaldi and a second
compound, ledipasvir, which inhibits the HCV protein
NS5A. The precise function of NS5A in the viral life
cycle is not known, however it is thought to play a role
in viral replication, assembly and secretion. The exact
mechanism of action of ledipasvir is not known, but it is
thought to bind to the NS5A protein and inhibit its role
in the HCV life cycle.

MIXING IT UP WITH VIEKIRA PAK

The end of 2014 gifted yet another breakthrough
therapy approval for HCV—AbbVie’s Viekira Pak.
A cocktail of three new drugs, Viekira Pak is also
a significant improvement over interferon and
ribavirin treatments.
• The first component of Viekira Pak, dasabuvir,
is a non-nucleoside analog inhibitor of the viral
polymerase. It simply binds to the virus in such
a way that it creates a conformational or shape
change to the enzyme itself, rendering it ineffective.
• The second component, paritaprevir, inhibits the
viral protein NS34A, which is critically important in
the HCV lifecycle and is a protease, or an enzyme
that “cleaves” other proteins. When HCV infects liver
cells, it hijacks the cell’s protein making machinery
for its own use. One of the proteins produced • The third component, ombitsavir, inhibits the HCV
is a long “polyprotein,” or protein composed of protein NS5A, as described above for the ledipasvir
many smaller proteins. In order for those smaller component of Harvoni.
proteins to do their job, they must be released Viruses with high mutation rates are unlikely to develop
from the polyprotein and the NS34A protease does mutations that confer resistance to all three drugs at
so by cutting the polyprotein into smaller bits. once, and treating them with a drug cocktail such as
Paritaprevir works by binding the part of NS34A Viekara Pak can be a successful strategy.
that does the actual cutting, preventing it from
doing its job.

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THE STATE OF CYSTIC FIBROSIS
AND PRECISION MEDICINE Life Science Training from Industry Experts

During President Obama’s State of the Union address production of sweat, digestive fluids and mucus. It
last month, a cystic fibrosis patient named Bill Elder sat affects around 70,000 people globally and is prevalent
beside First Lady Michelle Obama. Diagnosed with the in America, Europe and Australia.
disease at 8 years old, Mr. Elder is ”healthier now than The CTFR protein is classified as a channel protein—a
ever before” at age 27, thanks to Vertex’s (Boston, MA) category of proteins that create a channel, or tunnel,
Kalydeco. As a third-year medical student, he is not only across the cell membrane. This specialized gateway
surviving, but thriving. Receiving an invitation to be the allows things to pass through the cell that will otherwise
guest of honor at the presidential speech of the year is be denied entry or exit.
the exclamation mark to an extraordinary story.
Negatively charged chloride ions use CTFR to exit cells,
Mr. Elder is an example of the success of modern and if CTFR is not functioning correctly, the chloride
medicine. His cystic fibrosis (CF) treatment derives ions build up inside of cells. The build-up affects the
from an understanding of the underlying molecular fluid balance of tissue, which results in characteristically
causes of the disease. This approach, referred to by thick mucus seen in the lungs of CF patients. This thick
the President as precision medicine, is the focus of new mucus makes CF patients vulnerable to potentially fatal
federal investments to speed the development of lung infections.
targeted therapeutics—drugs designed for a subset of
CF is an autosomal recessive disorder, meaning if
patients with a specific genetic defect rather than for
an individual has one functioning copy of the CTFR
the “average” patient.
gene, they are termed “carriers” and will not develop
In this WEEKLY we’ll take a closer look at precision the disease. Two copies of the malfunctioning CTFR
medicine as it applies to cystic fibrosis. gene, one from each parent, will equal a diagnosis.
And while CF is always caused by a mutation, many
PERSONALIZED MEDICINE possible mutation combinations have been associated
VS. PRECISION MEDICINE with the disease–up to 1500 mutations, maybe more,
Personalized medicine implies the development of are possible.
medicines for an individual, based on their unique
Precision medicine plays the hero by identifying the
genetic, metabolic, microbiomic and other “signatures”.
exact effect these underlying mutations have on CTFR,
Think of a breast cancer patient getting a genetic test
and designing treatments to overcome the disease.
for the BRCA gene to determine their specific genetic
mutation and subsequent personalized course of ON THE MARKET
treatment—not just a therapy for all BRCA-induced
Cystic Fibrosis is symptomatically managed by reducing
cancers. As large scale, full-genome sequencing
the risks of lung infections and implementing lifestyle
becomes more efficient and common, we may start to
changes to prevent such infections. Antibiotics
see truly personalized medicines.
are taken at the slightest sign of sickness, or even
But for now, a better term is “stratified”or prophylactically, and other medications work to thin
precision medicine—dividing patient groups into mucus. As the disease progresses, a double lung
specific populations and designing new drugs for transplant may be the only, albeit elusive, treatment.
those subtypes.
That dire medical outlook changed for a subset of CF
WHAT IS CYSTIC FIBROSIS? patients in 2012, when Vertex won FDA approval for
its small molecule drug Kalydeco. Kalydeco works by
Cystic fibrosis is a genetic disease caused by one of
binding to the misfolded CTFR channel protein and
several possible mutations in the gene encoding the
increasing its ability to remain open and functional on
“cystic fibrosis transmembrane conductance regulator”
cellular surfaces. Kalydeco is indicated for fewer than
(CTFR) protein. The CTFR protein is critical for the

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10% of CF patients; Mr. Elder is one of the lucky ones
who responds to Kalydeco.

ON THE HORIZON
With the success of Kalydeco under their belt, Vertex is
after a bigger share of the CF market with another small
molecule drug called Lumacaftor.
Currently in Phase III trials stateside, Lumacaftor is
paired with Kalydeco, and to target the most common
CF mutation responsible for ~70% of the diagnosed CF
cases in US caucasians. In these patients, the channel
protein is so damaged it never makes it to the cell
surface. Lumacaftor corrects some of the misfolds,
improving CTFR’s ability to travel to the cell surface.
In Europe, where ~70% of diagnosed CF cases
may benefit from Kalydeco with Lumacaftor, the
combination therapy has gained accelerated review.
Back in the USA, the biotech world awaits the results
from the trials for the combination therapy to be
announced this July.

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LONGEVITY IN BIOTECH Life Science Training from Industry Experts

Longevity has fascinated mankind for centuries, from REPAIRING OLD WITH NEW
ancient myths about fountains of youth to current
A technique known as “parabiosis”–surgically joining
speculation around caloric restriction.
two mice to share a circulatory system–demonstrated
With our increased scientific understanding of aging, exposure to younger blood enabled older mice to repair
clinical validation and commercialization of treatments damaged liver and muscle tissue, likely due to stem cell
are near. The goal is to extend lifespan as well as health activation. Last year a Stanford research group, led by
span, and we may be looking at a future where the Tony Wyss-Coray, demonstrated old mice exposed to
norm is to remain mentally and physically active at age the blood of young mice have increased neuron growth.
90, or even 100. Wyss-Coray didn’t use parabiosis, rather he injected the
Healthy aging is influenced by a complex mixture of plasma component of blood from younger mice into the
factors, including genetics, lifestyle, environment and older mice, replicating the same effect; good news for
nutrition. Teasing out and integrating these factors into potential human patients.
a bigger picture is a major piece of the longevity puzzle. The encouraging mouse studies prompted Wyss-
While still in the experimental stages, the world of Coray to start Alkhest (Menlo Park, CA). In September
biotech is connecting the pieces to find viable pathways, 2014, Alkhest started Phase I clinical trials, testing
so let’s take a look at the players in the game. the infusion of plasma from young donors (under 30)
into Alzheimer’s patients (over 50). The company’s
THE LONG AND SHORT OF IT quick move into human studies is thanks to plasma
The connection between telomere length and aging transfusions being a common and safe practice,
gives us a glimpse into one of the many facets allowing the IND requirement to be skipped. The full
of longevity. speed ahead approach allows the company to rapidly
Telomeres are lengths of DNA, perched at the end of determine whether the technique is efficacious in
all chromosomes, made up of the repeating sequence people. Long term, isolating the exact protein(s)
of six nucleotides: TTAGGG. Our telomeres get enabling the beneficial effect is a likely next move
shorter as we age, especially in cell types that lack an for Alkhest.
active telomerase enzyme, whose job is to ensure the
entire telomere gets copied. A prevailing hypothesis SIMPLE TO COMPLEX
is telomere length can predict life expectancy, with Longevity research spans the relatively low-tech idea of
shorter telomeres corresponding to shorter lifespan. plasma transfusions to big data genomic sequencing,
Earlier this week, researchers at Stanford University evidencing its complexity.
(Palo Alto, CA) induced cells to transiently produce Human Longevity Institute (HLI; San Diego, CA),
telomerase by delivering a modified telomerase-coding established in 2014 by Craig Venter, plans to tackle
mRNA molecule to human muscle cells–resulting in a questions associated with longevity via human genome
significant extension of the telomeres. This transient sequencing–with a near term goal of 40,000 entire
expression is a key advantage because unchecked genomes per year and long term 100,000 genomes
telomere extension can cause cells to become per year. While many of the existing large-scale, full
cancerous. For now, the method is only used in the lab genome sequencing projects focus on cancer, HLI plans
to increase the lifespan of cells, but it may have future to sequence a diverse collection of genomes to tease
clinical potential as an anti-aging treatment in humans. out health and longevity patterns. Plans to mine the
Also undertaking the endeavor is Sierra Biosciences microbiomes and metabolomes of a sequenced group
(Reno, NV), working to identify compounds capable of subset are also in the works.
activating telomerase for clinical application.

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 lifespan”, Calico has been fairly secretive. Last fall’s
announcement of a potential $1.5 billion, 10–year deal
with AbbVie (North Chicago, IL) lifted the veil a little,
suggesting a plan to move swiftly into the clinic on
neurodegenerative diseases and cancer.

FROM NONTRADITIONAL
TO TRADITIONAL
Still others are approaching longevity from a more
traditional drug discovery approach–analyzing
pathways and identifying corresponding drug targets.
One such target is a “jack of all trades” enzyme known
as mTOR, which regulates the upward communication
from amino acids, growth factor and insulin; it
Another longevity company backed by legendary also monitors cell critical nutrients, energy levels,
players is California Life Sciences (Calico; South San and oxygen.
Francisco, CA), founded in 2013 with financial backing
Defects in mTOR signaling are linked to a wide range
from big data giant Google. Arthur Levinson, who
of age associated diseases including type 2 diabetes,
made his name first as head of R&D and subsequently
Alzheimer’s and rheumatoid arthritis. A FierceBiotech
CEO and chairman of Genentech (South San Franciso,
2014 “Fierce 15” company, Navitor Pharmaceuticals
CA), is CEO of the startup. Apart from a stated mission
(Cambridge, MA) is on the mTOR bandwagon, with aims
“to harness advanced technologies to increase
to develop proprietary technology to modulate mTOR in
our understanding of the biology that controls
various disease states.

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DESIGNER GENES: AN INTRODUCTION
TO GENOME-EDITING Life Science Training from Industry Experts

Five years ago, altering an individual’s genome would CATCHING THE RIGHT BREAK
have been labeled unimaginable. Fast forward to today
The secret to genome-editing lies in creating DSBs in
and one of the hottest new developments in biotech
therapeutically useful locations with a manmade enzyme
is genome-editing—the ability to selectively disable or
called a zinc finger nuclease (ZFN).
edit the sequence of specific genes.
How are ZFNs made? To start, zinc finger proteins
In this WEEKLY we will compare and contrast the
(ZFP) are sequence-specific, DNA-binding proteins
different genome-editing platforms in development and
that activate gene expression. They are engineered
discover how close we are to a clinical reality.
to recognize unique sites within the genome. While
GOT GAPS IN YOUR GENES? ZFPs do not have the ability to cut DNA on their own,
scientists can fuse a ZFP together with a DNA-cutting
Genome-editing is possible thanks to damage in the
enzyme called nuclease—the “N” in ZFN. The marriage of
DNA sequence and the cell’s subsequent activation
ZFP to nuclease creates ZFN.
of different repair pathways. These DNA breaks are
named double-stranded breaks (DSB) because both How do ZFNs work? ZFNs create the desired sequence-
strands of the double-stranded DNA helix are broken, specific, double-stranded break within the genome.
similar to a two lane bridge that has a section break off The DSB activates the NHEJ repair pathway, resulting in
into the water below after experiencing an earthquake. disruption of the gene, if there is a desire to “knock out”
that gene. If a repair template is delivered at the same
Repair pathways are charged with fixing the break:
time as the break, the HDR pathway kicks in.
• Non-Homologous End-Joining (NHEJ) closes the gap
between the break by joining the two sections
back together—imagine pushing the two sides of
the bridge together, leaving the fallen section in
the water. An unintended byproduct of NHEJ is
the possibility of sequence error, much like the
sections of the bridge not lining up properly; even
a single base deletion may cause unintended
consequences. If the repair occurs in the middle of
a gene, the minor error can be enough to disrupt
gene function and halt the production of the
corresponding protein.
• Homology Directed Repair (HDR) relies on a highly
similar (homologous) DNA segment to repair the
break - imagine the missing bridge section built
elsewhere and helicoptered in to fill the break. CATCHING AN EASIER BREAK
How does genome-editing start? Double-stranded The biotech world is abuzz with the promise of a new
breaks are engineered to occur at specific locations, technology with potentially greater flexibility called
activating the intrinsic cell repair pathways: HDR clustered regularly interspaced short palindromic repeats/
and NHEJ. CRISPR-associated 9 (CRISPR/Cas9).
CRISPR/Cas9 is similar in concept to ZFN genome-
editing; however, with CRISPR/Cas9, the nuclease
enzyme Cas9 is escorted to the correct location by
guide RNAs—or short sequences of RNA complementary

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to the target sequence. Since producing a new RNA
sequence is simpler than producing an engineered
protein, targeting the editing site is much easier and
faster using the CRISPR/Cas9 system.

CLOSING THE GAP IN GENOME EDITING

ZFN genome-editing is in Phase II trials for treatment of
the human immunodeficiency virus (HIV). HIV destroys
the immune system by infecting T-cells—a type of
white blood cell critical to mounting a defense against
invading pathogens. It turns out T-cells function just
fine when missing a protein called CCR5, which contains
a receptor primarily used by HIV to infect T-cells.
Sangamo Therapeutics (Richmond, CA) is using its ZFN
genome-editing platform to disrupt the CCR5 gene on
CRISPR/Cas9 has yet to enter clinical development
patients’ T-cells.
but several companies, including Editas Medicine
(Cambridge, MA) and Caribou Biosciences (Berkely,
CA), are in preclinical development of CRISPR/Cas9
technology indicated for a variety of genetic and
infectious disease targets.

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BIOSIMILARS: READY OR NOT, HERE THEY COME Life Science Training from Industry Experts

Last week, the FDA’s Oncologic Drugs Advisory Committee

unanimously recommended Novartis’ biosimilar
version of Amgen’s megahit Neupogen (filgrastim),
effectively paving the way for the first US biosimilar
approval in the near future.
Filgrastim is a recombinant version of a naturally-
occurring protein which stimulates the bone marrow
to increase production of white blood cells. Patients
undergoing chemotherapy, as well as those receiving
bone marrow transplants, are often prescribed
filgrastim to replenish the infection-fighting white
blood cells destroyed by these treatments. While
the European Medicines Agency (EMA) has already
approved seven versions of the drug starting back in
2006, the FDA is playing catch-up while it determines
need to carry out functional studies in the lab to show
the final parameters of the biosimilar launch.
their biologic drug is highly similar to the original, hence
Ready or not, biosimilars are making their the term biosimilar.
stateside debut.
PROTEIN PRIMER OVERSIMPLIFIED
EASILY CONFUSED:
The goal of a gene sequence is to code for an amino acid
BIOSIMILAR VS. GENERIC
sequence. The linear amino acid sequence then folds
A biologic is the product (typically a protein) of a living into a complex, three-dimensional shape to become a
organism, or cell, used to treat or manage a disease. functional protein.
How are biologic drugs made? By inserting the gene for
Of course, it’s not so simple: sometimes sections
a therapeutic protein into a bacterial or mammalian cell
of amino acids are cut out; sometimes sugars are
- which then makes the protein by following the recipe
added; sometimes proteins bind together. All of these
provided by the gene.
subsequent additions and subtractions affect the final
A biosimilar is more of a “remake” than a sequel. It aims folding pattern, or native protein shape. And in the
to treat or manage the same disease as if it were the world of proteins, shape is paramount in endgame
original biologic, but is produced using a different cell protein function. If the cell does not add, subtract,
line, master cell bank (MCB), and/or different process. or fold the protein correctly, the protein may not
The term generic drug refers specifically to copycat function properly.
versions of small molecule drugs. Because their To maintain uniformity in protein shape and function,
molecular structure is relatively simple, chemists can each therapeutic biologic is produced from a distinct
analyze the structure of a small molecule drug and working cell line, originating from a specific MCB.
synthesize it in the lab. Commonly available analytic
techniques are then used to demonstrate their product THE PRODUCT IS THE PROCESS
is identical to the original.
The gene sequences for therapeutic proteins
In contrast, the structure of a biologic is highly complex. are publicly available, so in theory it should be
The analytical technology of today is unable to straightforward to make a “copycat” version of any
demonstrate two proteins are 100% identical to each biologic drug.
other. Thus, a company making a follow-on version may

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In practice, however, many external factors can generic manufacturer may use the safety and efficacy
influence the final, intricately folded, three-dimensional data generated by the brand company - a savings of
shape of any protein product. hundreds of millions of dollars.
Cell lines originating from different MCB’s are not Biosimilars must sing a different tune since they are
identical, even if both are Chinese hamster ovary cells only similar, not identical, to the original biologic.
- a biomanufacturing favorite. These slight variations in Biosimilar manufacturers are not (yet) allowed to
the cells themselves may impact folding and the final cite safety and efficacy data from the original clinical
product structure. studies, and must run their own trials as defined by
The precise manufacturing strategy the innovator their governing regulatory body.
uses is proprietary, so subsequent biomanufacturing How does the regulatory process work? At this time, the
protocols are likely to be a variation of the original. EMA (and now the FDA) allow biosimilar manufacturers
Thus, conditions under which the cells are grown also to present data, based on laboratory characterizations,
influence the final structure of the protein product. showing their proposed biosimilar is “highly similar”
These facts give rise to the saying “the product is the or “comparable” to the original product. Based on this
process” - which cannot be easily duplicated. level of similarity, the regulatory agency may then allow
abbreviated clinical studies. In other words, biosimilar
ONE OF THESE CELLS IS manufacturers must still test for safety and efficacy
NOT LIKE THE OTHER in human patients, however, the amount of testing
Because the chemical structure of the generic small required varies on a case-by-case basis and carries less
molecule drug is identical to the original drug, the burden than a new approval.

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THE BISPECIFIC ANTIBODY: A LETHAL HYBRID Life Science Training from Industry Experts

Amgen’s acute lymphoblastic leukemia treatment, on the surface of blood cancer cells, and the other
Blincyto, coasted past the FDA bottleneck ahead of arm targets CD3, a protein that activates the T-cell
schedule, gaining approval earlier this month as a receptor complex.
“first of its kind” immunotherapy. The FDA designated
Blincyto as a breakthrough therapy and an orphan
product on top of granting a priority review, thus
propelling the bispecific antibody (BsAb) to the market
five months early.
The swift approval and technology behind this
monoclonal antibody (mAb) hybrid caught our eye
here at WEEKLY, so let’s break down the science
behind Blincyto.

TERM OF THE WEEK:

BISPECIFIC ANTIBODY
A bispecific antibody is a genetically engineered
The above image represents the two arms of a BsAb.
protein composed of two different monoclonal
antibody fragments.
Bringing the CD19 and CD3 together ensures killer
The descriptor bispecific derives from mAb’s unique T-cells are triggered to target the cancer cells. Activation
ability to bind two different types of antigens. also induces T-cell proliferation and the descendants of
Traditional monospecific mAbs have affinity for only the activated T-cell also target the tumor cells.
one target. A bispecific antibody has specificity for
a target cell on one arm and a therapeutic (or killing MARKET WATCH:
agent) on the other. BISPECIFIC ANTIBODIES
The European Medicines Agency is ahead of the curve
KILLER T-CELL ACTIVATION with two BsAb approvals under their belt. Neovii
Cytotoxic T-cells (more commonly referred to as killer Biotech’s (Grafelfing, Germany) Removab fights
T-cells) can be thought of as the warriors of the immune malignant ascites—a fluid buildup as the result of a
system. They compose the front-line of the specific tumor. Active Biotech’s (Lund, Sweden) Anyara knocks
immune response, activated to attack foreign invaders. out cancerous cells in renal cell carcinoma.
Normally, this activation occurs when a unique T-cell
With the stateside debut of BsAbs underway, plenty
receptor recognizes a specific target protein.
of other companies are looking to jump into the mix:
Once activated, cytotoxic T-cells can become highly TRION Pharma (Munich, Germany), Amgen (Thousand
effective cancer killing machines. There is a catch: Oaks, CA), AbbVie (North Chicago, Illinios), Ablynx
T-cells often miss tumor cells because they are not (Ghent, Belgium), Affimed (Heidelberg, Germany),
considered by the body to be foreign. The drug and MacroGenics (Rockville, MD) all have BsAbs in
discovery challenge lies in figuring out how to create an development. While the majority of the BsAb pipeline
army of tumor-specific T-cells. targets cancer, about 25% go after rheumatoid arthritis,
other autoimmune disorders, and respiratory disease.
BLINCYTO: MECHANISM OF ACTION
Blincyto redirects killer T-cells to target specific tumor
cells. How? One arm grabs CD19, a protein found

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HITTING THE MARK WITH ALZHEIMER’S
DRUG DEVELOPMENT Life Science Training from Industry Experts

The biotech world is still aiming to stop Alzheimer’s To begin, the amyloid precursor protein (APP) is
disease despite many recent drug attempts missing characterized as transmembrane—spanning the entire
the mark in late phase clinical trials. While the effects membrane—and is found in the synapses of neurons.
of Alzheimer’s are well known (the degeneration of the APP is thought to play a role in the formation of
brain over time due to nerve cell death), the cause of synapses, or the gaps between neurons through which
the disease remains a mystery. chemical messages pass.
As people live longer worldwide, the incidence of Aβ plaques derive from the cleavage of APPs. Aβ is
Alzheimer’s continues to increase, and the current released by the cleavage of the extracellular portion of
approach only manages symptoms. With new hope APP via the enzyme beta-secretase 1 (BACE 1). Then, the
from Biogen Idec (Cambridge, MA) alongside other individual Aβ molecules subsequently clump together
encouraging pathways in drug development, the quest to form the plaques associated with Alzheimer’s.
for a cure remains strong. Last year researchers at Stanford School of Medicine
In this issue, we will touch upon what is known about (Stanford, CA) demonstrated that Aβ binding to a
Alzheimer’s and circle in on the current state of drug receptor on nerve cells disrupted the function of the
discovery and development. synapse. This may be the mechanism by which the Aβ
plaques cause Alzheimer’s. The ability to interrupt this
CIRCLING IN ON THE interaction could potentially preserve functioning nerve
AMYLOID HYPOTHESIS cells and points the way to potential drug targets.
Amongst a sea of alternative theories, the current Another approach to Alzheimer’s drug development
consensus points to Alzheimer’s disease being involves the inhibition of Aβ production in the first
associated with the buildup of beta-amyloid plaques (Aβ) place. Several big players—such as Lilly (Indianapolis,
in patients’ brains. What exactly are Aβ plaques? IN), AstraZeneca (London, England), and Biogen Idec—
are sharpening their arrows with hopes of making it on
the board.

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TARGETING TAU PROTIENS Other companies have attempted to develop mAb
therapies for Aβ and ended up abandoning ship. Why is
Currently, most Alzheimer’s therapies in development
Aβ still being pursued as a drug target? Different mAbs
target Aβ plaques; however, some researchers have
attach to different parts of Aβ and the outcome of one
their eye on another brain protein called tau.
mAb trial does not necessarily predict the outcome
Tau is concentrated in the neurons and is primarily of another.
understood as a component in stabilizing nerve cell
structure. Abnormal aggregates of tau form tangles GENETICS PLAYS A ROLE
within nerve cells; they are correlated with the About 70% of Alzheimer’s cases are thought to have
onset of Alzheimer’s along with the characteristic at least some genetic association, with different genes
plaque formations. being implicated depending on the type of Alzheimer’s.
Naturally, a tau-aggregation inhibitor seems like a A gene found on chromosome 19 called the
good therapeutic to test, and Singapore-based TauRx apolipoprotein E gene (APOE) influences the
is amidst late stage clinical trials with a developmental development of late-onset Alzheimer’s. Individuals
drug targeting the tau pathway. with different variants of the APOE gene have different
risk profiles:
MONOCLONAL ANTIBODIES
MAKE THEIR MARK • Variant ε2 (APOE2) is rare and possibly lessens or
delays Alzheimer’s onset.
Last week Biogen Idec announced positive results in
early stage clinical trials of developmental drug BIIB037, • Variant ε3 (APOE3) is neutral.
a monoclonal antibody (mAb) targeting Aβ and sights • Variant ε4 (APOE4) is associated with a significantly
are set on Phase III. increased risk of Alzheimer’s.
Investigators reported time-dependent and dose- The APOE proteins plays a role in clearing Aβ from
dependent reductions in Aβ levels. The more of the the brain, with APOE4 carrying out this function less
drug given to the patients and the longer they were efficiently than the other variants. There is also some
exposed, the greater the reduction in Aβ—the gold evidence that APOE4 contributes to the breakdown
standard for Alzheimer’s drug efficacy studies. BIIB037 of the blood-brain barrier seen in patients, resulting
demonstrating time and dose dependency makes it in increased brain inflammation—another marker of
much less likely that the reduction of study subject’s Aβ Alzheimer’s. A better understanding of APOE4’s role
levels is caused by other factors. in Alzheimer’s onset may lead to the development of a
whole new class of drug.

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CAPTURING THE RUNNING BACKS OF CANCER Life Science Training from Industry Experts

After using diagnostic technology to establish the

existence of cancer, determining the severity is the next
step toward developing a game plan. If metastasis is
in the picture, strengthening your defensive line-up is
a must.
The dissemination of circulating tumor cells (CTCs) from
the primary tumor to unaffected tissue is a key step
of early stage metastasis. CTCs break away from the
original tumor and migrate to other areas of the body
via the bloodstream. Metastasis is the main player
propelling cancer to spread throughout the body.
There is conflicting evidence on whether invasive
procedures that aim to diagnose and cure can
potentially trigger metastasis. Regardless of the debate,
the need to stop cancer at various stages is perpetually attraction to a protein called stromal-cell derived factor
in demand. 1 or SDF-1.
In this issue of WEEKLY, we will piggyback from Preclinical work by scientists at Johns Hopkins
our earlier look at CTC diagnostics and examine University (Baltimore, MD) suggests implanting a
developmental devices aimed at capturing CTCs—the device coated in SDF-1 could help to capture migrating
running backs of cancer. Some of these clever devices prostate cancer cells. A similar idea is being explored
have the potential to be used in conjunction with by researchers at Santiago University (Santiago, Chile)
a cancer biopsy or surgery to prevent the possible with a technology called M-trap. M-trap would be
escape of tumor cells during an otherwise life- implanted inside the abdominal cavity of ovarian cancer
saving procedure. patients to seize migrating cancer cells. Both types of
implantable devices have the potential to tackle the
EASILY CONFUSED: BENIGN threat of metastasis.
VS. MALIGNANT TUMOR
A benign tumor does not invade its surrounding TACKLING CANCER WITH MAGNETS
tissue or spread around the body. It stays contained Magnets developed at the University of Arkansas
within the original site. A malignant tumor may invade (Fayetteville, AR) and subsequently licensed to bay area
its surrounding tissue or spread around the body, biotech Accurexa, punt an elegant approach into the
wreaking havoc on its path to metastasis. arena: a treatment that sidesteps surgery (if proven
successful in clinical trials).
PLANTING AN M-TRAP
Magnetic nanoparticles covered with tumor-cell-
One way to pump up the defensive line is to implant specific antibodies are injected into patients, then the
physical traps to attract and capture tumor cells as they nanoparticles get to work and mop up CTCs attracted to
start to emerge from the pack. the tumor-cell-specific antibody coating.
Different types of primary tumor cells are attracted The innovation comes into play in a rather simple
to specific proteins - those primary tumor cells tend fashion—a magnet is held to the patient’s skin,
to spread to tissues where those specific proteins attracting the CTC-bearing magnetic nanoparticles.
are concentrated. For example, breast and prostate A laser is then beamed at the huddle, heating up the
cancer cells tend to spread to the bone because of their

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magnetic nanoparticles and destroying the attached FDA approved Mozobil (Sanofi, New York, NY) makes
tumor cells. a run for the end zone by flushing bone marrow stem
This promising technique has only been tested in cells out of cancer patients. Mozobil blocks the cells’
animal models, but clinical trials are being worked interaction with SDF-1—the protein that attracts
out for breast cancer patients to include magnetic cancer cells to the bone marrow in the first place.
nanoparticles as a component of their surgery. When treated with Mozobil, prostate cancer that had
spread to bone marrow appears to push those cancer
BLOCKING THE PLAY WITH SDF-1 cells back into the bloodstream, making them more
susceptible to other types of therapy. Mozobil could
The ideal scenario is for cancer cells to never make it
potentially be used in combination with some of the
past the starting line, but what if they have already
highlighted devices above, adding two superstars to the
made their way down the field?
dream team.

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12 BIOTECH CONCEPTS EVERYONE
SHOULD KNOW Life Science Training from Industry Experts

Our mission here at WEEKLY is to keep your industry

knowledge up to date. We will be off Thursday,
roasting our turkeys and reminding ourselves that
it is not the amino acid tryptophan that makes us
drowsy after a Thanksgiving feast—it is the massive
carbohydrate intake!
Whether you are awaiting a Thanksgiving holiday or
it is business as usual, we have compiled 12 biotech
concepts for you to browse.

BIOTECH BASICS
• Learn about the darlings of biotech:
monoclonal antibodies.
• What is up with gene chips?
DISEASE PROFILES
DRUGS IN DEVELOPMENT
• ALS ice bucket challenges have almost frozen over,
• PD-1 and PDL-1 are hot topics, discover more about but the science still applies.
check point therapies.
• Celiac Disease sufferers want a slice of biotech cake.
• What is going on with drugs in development for
• How does Ebola wreak such havoc?
your heart?
• Read how the powerhouse mitochondria opens up
a possible pathway.

FDA APPROVED THERAPIES

• Find out about the buzz behind inhalable insulin.
• What are the mechanisms of action for Abilify,
Nexium, and Humira?

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PIONEERS IN THE LAND OF SMALL PROTEINS Life Science Training from Industry Experts

We continue last week’s trek into the IPO wilderness to

explore the technology of Molecular Partners (Zurich,
Switzerland). What they are in search of is a new class
of small protein therapeutics, referred to by their
acronym: DARPins.
DARPins inherently share some of the same advantages
monoclonal antibodies (mAbs) have over small molecule
drugs. DARPins go a step further and offer possible
upsides to some known mAb downsides.
The potential of DARPins—the driving force behind
Molecular Partners—shows promise enough for
a planned fourth quarter debut on the Swiss SIX
exchange. Let’s strap on our hiking boots and unearth
the science behind DARPin technology.

PROTEIN PRIMER
DARPins can be better understood in the context of
a primer on proteins. Proteins are complex, three-
dimensional formations built up from intermediate
layers of structure. The most basic level of protein
structure is the primary structure, or the linear
sequence of the amino acid building blocks that make
up the protein:

Source: Wikipedia
The next level, secondary structure, consists of highly
regular three-dimensional structures formed by
Regions of secondary structure may be grouped
interactions between neighboring amino acids. Two
together to form a motif. Motifs are patterns found in
common types of secondary structure are alpha helices
several different proteins.
and beta sheets:
The compact globe-shaped configuration formed by the
interactions of alpha helices and beta sheets is called
the tertiary structure. In many functional proteins,
the final formation consists of two or more tertiary
structures that fit together to form one protein, termed
the quaternary structure.

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WELCOME TO THE LAND OF DARPINS following table illustrates, small and simple may be
an advantage.
The moniker DARPin stands for Designed Ankyrin
Repeat Protein. An ankyrin repeat is a motif that
consists of two helices separated by a loop, as shown
below. The number of helix-loop-helix repeats can vary,
ranging from four to thirty-four.

Source: Wikipedia CLINICAL TRAILS: DARPINS

In nature, the ankyrin repeat motif mediates a wide The first DARPin therapeutic, Molecular Partners’
variety of protein-to-protein interactions. The number Abicipar, has ascended up the FDA mountain to Phase III
of repeats influences the final configuration of the motif clinical trials for wet age-related macular degeneration
and this final shape determines which target protein the (AMD).
ankyrin repeat domain will influence. Wet AMD is caused by excessive blood vessel growth
Using genetic engineering, scientists at Molecular to the retina, potentially leading to retinal detachment.
Partners are creating a large library of ankyrin repeat Abicipar binds to and inhibits the growth factor, known
proteins, hence the design aspect of Designed Ankyrin as vascular endothelial cell growth factor (VEGF), that
Repeat Proteins. These DARPins are screened for activates the excessive blood vessel growth.
their ability to interact with and modulate a variety Molecular Partners continues to expand its territory
of drug targets, with the prospect of being used in with a multi-VEGF/PDGF DARPin in preclinical
future therapies. development. This DARPin targets both VEGF and a
second growth factor: platelet-derived growth factor
DARPINS VS. (PDGF), also implicated in wet AMD. Scouting reports
MONOCLONAL ANTIBODIES predict the ability to target two different growth factors
All of this talk about DARPin technology begs the will lead to even greater efficacy in clinical trials. Both
question: how exactly do these small proteins compare wet AMD therapies are being developed with support
to today’s biotech darling: mAbs? It is no accident that from Allergan (Irvine, CA).
seven out of the top ten best selling drugs are mAbs, On an encouraging endnote, other DARPins developed
also known as large molecule drugs—their target by Molecular Partners show preclinical and early clinical
specificity mitigates disease with few adverse reactions. success against solid tumors. We will be keeping watch
What is obvious when comparing the monoclonal on the field of DARPins as they make their way along the
antibody to the DARPin is size and complexity of drug development trail.
structure. MAbs are large and complex and as the

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DRIVING DOWN THE EPIGENETIC HIGHWAY Life Science Training from Industry Experts

proteins a cell will end up producing. Gene expression is

the reason a heart cell is different from a liver cell.
Gene expression differences are also associated with
disease. For example, a type of cell or tissue may make
too much or too little of a particular protein, which is
the basis for many genetic disorders.

BEYOND GENETICS: EPIGENETICS

Epigenetics are changes to DNA that do not alter
the actual gene sequence, but rather are chemical
modifications to the DNA itself. These changes typically
affect gene expression, or how the gene is read by the
cells. Epigenetic modification can occur either directly
to the nucleotide bases themselves (A, C, G, or T) or to
the histones, which are small proteins that package and
order DNA.
Biotechnology is enjoying another banner year on
Wall Street. Currently, there are at least 39 companies
queued for an IPO. This led us at the WEEKLY to wonder:
what is the actual science behind each company’s
promise that leads public markets to invest billions
of dollars? This week and next, we are popping open
the hood and checking out what exactly merits such
a milestone.
First up is Syndax (Waltham, MA), a biopharmaceutical
company pumping out a new epigenetic therapy
for cancers resistant to treatment, which pique our
interest because of the novelty of their approach.
In this issue, we will get behind the wheel and go for
a ride to examine epigenetics and its potential for
future therapies. The above image illustrates how histones are tiny
spheres in which DNA wraps itself.
TERM OF THE WEEK: One of the most common types of epigenetic
GENE EXPRESSION modification is methylation—the addition of a methyl
Genes are the actual DNA sequence encoding (CH3) group to cytosine (C) nucleotides. The result:
for a protein that confers a particular trait and methylation reduces or even blocks gene expression.
genetics is the study of how genes determine an A second type of modification is called acetylation—the
organism’s characteristics. addition of an acetyl group (CH3 CO) to the histones.
Gene expression is the process cells use to read genetic Acetylation loosens the association of the DNA with
information to make proteins. Because each cell in the histones, making the DNA more accessible to the
our body has the same genetic information, it is the enzymes used in gene expression, ultimately increasing
differences in gene expression that determine what protein production.

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Deacetylation—the removal of an acetyl group— THE FUTURE OF HDACS
increases the association or “grip” of the DNA around
HDACS are associated with more diseases than
the histone proteins. Deacetylation makes the DNA less
cancer, and Acetylon Pharmaceuticals (Boston, MA)
accessible to enzymes used in gene expression, thereby
is in preclinical development for HDAC inhibitors—
decreasing the production of proteins.
potentially treating autoimmunity, neurodegeneration,
CLINICAL TRIALS: ENTINOSTAT cardiac disorders, stroke, diabetes, and depression.
Their HDAC inhibitor-based myeloma therapy is also
Syndax’s new drug, Entinostat, is a histone deacetylase
in development.
(HDAC) inhibitor. HDACs are enzymes that increase the
expression of certain genes by deacetylation, or the As the field of epigenetics flourishes, an increasing
removal of acetyl groups from histone proteins. number of therapeutics targeting this mechanism
are expected to gain ground. Enzymes involved in
Entinostat’s goal is to target HDACs overexpressed in a
epigenetic modification appear to play a role in a
variety of cancers, potentially leading to the shutdown
range of different disorders. The true challenge will
of uncontrolled cell growth. Entinostat targets a
be in developing targeted inhibitors for each subset
common mechanism; perhaps it could one day be used
of enzyme.
in a range of different cancers if shown to be effective.
Currently, there are ongoing Phase II trials studying
the effect of Entinostat combined with Roche’s
Tarceva on Hodgkin’s lymphoma and metastatic lung
cancer. In addition, Phase III trials are underway for
studying advanced breast cancer, where Entinostat is
combined with Pfizer’s Aromasin. Notably, Entinostat
combined with Aromasin has received the FDA’s “New
Breakthrough Therapy” designation for breast cancer.

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BREAKING FROM THE PACK WITH
CANCER DIAGNOSTICS Life Science Training from Industry Experts

Regular WEEKLY readers know that cancer stems from CASTING A WIDER NET
the division of a single cell leaping to uncontrolled
One criticism of using an mAb-based approach to
growth and then growing into a tumor. How exactly
isolating CTCs. It forces the researcher to limit the
does cancer disrupt the life process? Sometimes tumors
specific cell-surface protein markers they choose to
grow to such a size that the function of a vital organ is
focus on during the isolation process. Not all CTCs
no longer viable. Mostly, cancer kills by metastasis—
carry the same markers and many have not even been
when cells from the tumor break away and spread to
identified, so finding a more universal marker is a
other unrelated tissues.
top priority. One such marker may be cell size—CTCs
Circulating tumor cells (CTCs)—cells splintered from a tend to be significantly larger than other cells in the
tumor and circulating in the bloodstream—reveal one blood, and this size differential may be exploited in a
of the earliest signs of metastasis. These renegade microfluidics-based approach to cell separation.
cells are the running backs of cancer. Detecting these
Researchers at National University in Singapore
cells early can lead to quicker diagnosis. Monitoring
(Singapore) and MIT (Cambridge, MA) have developed
these cells can result in tracking changes over time
a microfluidics chip that routes cells from a blood
without repeated invasive biopsies and highlighting
sample into different channels based upon cell size.
the diagnostic power of CTCs ability to assess the
The larger CTCs directed into the appropriate channel
effectiveness of various therapies in real time.
are easily collected for further analysis. Although still
The challenge lies in detecting CTCs; some estimates in the preclinical research phase, this approach shows
classify them as rare as one circulating tumor cell promise for capturing a wide range of CTCs.
per billion normal cells! With only one CTC diagnostic
currently on the market, the game is on for other COCKTAIL FODDER
companies to bring their players to the field.
CTCs are not new to the game of diagnostics. We
have known about them since 1869 when pathologist
THE SEARCH IS ON
Thomas Ashworth observed them in the blood of a
Janssen Diagnostics (Raritan, NJ) currently markets cancer patient. Researchers finally honed in on the
CellSearch, the single FDA approved test that allows technology in the 1990s and CellSearch was granted
physicians to identify early CTCs from blood samples. FDA approval in 2004.
How does CellSearch work? Monoclonal antibodies
(mAbs) capable of recognizing proteins on the surface of NO CELL LEFT BEHIND
migrating tumor cells are chemically linked to magnetic Epic Sciences (San Diego, CA) adopts a “no cell left
nanoparticles and then added to a patient’s blood behind” game plan thanks to technology developed by
sample. These tumor-specific mAbs grab hold of the the Scripps Research Institute (La Jolla, CA).
CTCs and a strong magnetic field is then applied to the
Automated fluorescence-microscopy identifies the
sample, isolating the captured cells for identification
CTCs in blood samples placed on microscope slides.
and analysis.
A detailed analysis of three million cells per slide is
A higher number of CTCs detected may indicate a performed, each blood sample yielding approximately
higher incidence of metastasis, or a less than effective twelve slides. This technology may potentially hone in
treatment route if used to quantify cancer therapy on the presence of a single CTC.
success. CellSearch keeps tabs on CTC cell counts for
Epic Sciences currently uses their test to perform
breast, prostate, and colorectal cancers.
analyses for biotech, pharmaceutical, and clinical
research partners with a long term goal of releasing a
diagnostic product for reference labs.

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BIOTECH’S BATTLEFRONT:
MONOCLONAL ANTIBODIES Life Science Training from Industry Experts

Since their premier on the scene, monoclonal The mAbs that use immune system activation as a MOA
antibodies (mAb) have demanded top billing on the in their portfolio include Roche’s Herceptin for breast
biotech marquee, creating a cast of therapeutics cancer and Biogen Idec’s Rituxan for lymphoma.
used to treat diseases like autoimmune disorders and
cancer. The first player debuted in 1986 when Janssen- THE GREAT COVERUP
Cilag’s OKT3 gained FDA approval to treat transplant Receptor proteins (proteins residing on the cell surface
rejection patients. that receive external signals) lose their ability to
Fast forward to 2013, where half of the top ten best internally transmit those external signals when covered
selling drugs worldwide happened to be mAbs: up by mAbs. Blocking signaling pathways effectively
Avastin, Herceptin, Humira, Remicade, and Rituxan. work as a form of treatment in cancers where growth
And do not hold back the applause because the FDA factor receptor activation plays a key role in turning
approved four new mAbs this year. Eli Lilly’s Cyramza on cell division. Blockers of receptor signaling include
takes a punch to stomach cancer. Janssen’s Sylvant Amgen’s Vectibix for colorectal cancer and Bristol-
is reigning in Castleman’s disease—a rare disorder Myers Squibb’s Yervoy for melanoma.
similar to lymphoma. Takeda’s Entyvio delivers a double
knockout, treating both ulcerative colitis and Crohn’s
disease. Rounding out the end of the pack is Merck’s
Keytruda, putting the blinders on melanoma.
Here at WEEKLY, we marvel at the multifaceted
mechanisms of actions (MOA) of mAbs as they have
evolved over the years. In this issue we will go behind
the scenes and find out exactly how they manage to
The Y shaped mAb above blocks the transmission of external signals.
shine the light on treating disease.
mAbs also cover up signaling molecules directly by
THE FRATERNAL TWIN capturing the signaling molecule itself and tethering it
The fraternal twin to the therapeutic mAb is the down before it can reach the receptor. Roche’s Avastin
naturally occurring antibody, which continually marches is roping in a variety of cancers while GlaxoSmithKline’s
to the frontline to defend the human body against Benlysta is tamping down the out of control immune
foreign invaders. When administered, the therapeutic reaction in lupus.
mAb acts as a freshly recruited soldier that alerts and
works with the immune system to destroy target cells STOP, WHO GOES THERE?
such as pathogens, cancer, or other diseased cells of Meet the sentry mAbs, denying infectious pathogens
the body. from gaining entry to the portals of their target cells.
The life goal of pathogens are to gain entry in to the
host via attachment to the surface of the target cell.
The link between pathogens and host cells can be
prevented, courtesy of a mAb attaching to the area
on the pathogen that interacts with the host cell,
blocking admittance.

The Y shaped mAb above alerts the immune system to send a macrophage.

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 bound, the deadly payload is internalized and delivered
to the innards of the tumor cells. This trojan horse
treatment is less toxic because it is only delivered to the
cancer cells, leaving the neighboring healthy cells in the
patient’s body relatively unharmed.

The virus above attempts to enter the host cell

but is denied, thanks to a mAb.

Astra Zeneca’s Synagis is guarding the gates against

respiratory syncytial virus. It prevents viral infection by
binding to proteins on the surface of a respiratory virus,
stopping the potentially lethal virus in its tracks.
The toxic “drug bomb” attached to the mAb above
successfully annihilates the target cell.
TROJAN HORSE
While several antibody-drug conjugates are in the midst
Increasingly, mAbs are being used as couriers to
of development, only two antibody-drug conjugates
deliver a toxic drug to target cells. They are known as
have made it to market. Roche’s Kadcyla is bumping out
antibody-drug conjugates. In this application, a highly
breast cancer while Seattle Genetics’ Adcetris mutes the
toxic compound is chemically attached to an mAb that
cancerous lymphocytes in lymphoma.
recognizes proteins on the surface of a cancer cell. Once

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PURSUING THE PROMISE OF
UNLIMITED PLATELETS Life Science Training from Industry Experts

Universal platelet cells—generated from human TERM OF THE WEEK:

induced pluripotent stem cells—made their debut HISTOCOMPATIBILITY
last week, courtesy of Advanced Cell Technology
The prefix “histo” is derived from the Greek word
(Marlborough, MA). The possibility of producing
for tissue, so histocompatibility simply means tissue
platelets—the component of blood that stops bleeding
compatibility and is often used in the context of organ
on-demand and circumventing donor rejection—could
or tissue transplants.
be a game changer for both platelet transfusions and
even other tissue transplants. Histocompatibility is determined by the MHC proteins
on the surface of cells and tissues. It appears there are
Universal platelets are free of the infectious agents
six different proteins that determine compatibility; the
found in donated versions, allowing companies to skip
higher the number of identical proteins, the greater
the mandatory steps of screening donors and testing
the chance an organ transplant will be successful. Only
blood. Platelets also lose their value over time—they
identical twins share all six proteins in common.
cannot be frozen, unlike their plasma and red blood
cell counterparts that make up blood. They only last
at room temperature for up to five days and are more
COCKTAIL FODDER
likely to be in short supply. The tantalizing idea of mass Do not ever expect to see aspirin setting up romantic
produced universal platelets pinged our radar here dates between platelet cells because it is a known anti-
at WEEKLY. In this issue, we will examine the science coagulant—that is, it interferes with the body’s ability to
behind the potential of universal platelet technology. form clots. Patients at risk for heart attacks and strokes
are often prescribed low-dose aspirin therapy to reduce
AXING THE MHC PROTEIN the risk of these lumpy platelet formations. The flip
side is that stopping internal bleeding may prove to be
Major histocompatibility (MHC) proteins reside on
difficult with patients on an aspirin regimen.
the surface of cells and tissues in our body, playing
an important role in activating the immune reaction Aspirin reduces inflammation by inhibiting a chemical
by alerting the immune system to the presence of called COX1, giving aspirin its pain-relieving and fever-
a foreign invader. Certain specialized MHC proteins reducing properties. The inhibition of COX1 also makes
actually bind to antigens and present them to T-cells platelets less sticky. In other words, they are less
for inspection, deciding whether or not to destroy the likely to clump together and initiate clot formation.
unwelcome entity. The protective effect lasts for as long as the platelets
exposed to the aspirin are in circulation, and up to
Organ donation rejection can occur when the MHC
five days after aspirin use has stopped. This prevents
proteins on the surface of a donor organ are not the
amorous platelet bonding in the near future.
same as the proteins on the surface of the recipient’s.
Platelet cells also have MHC proteins on their
surface, which come into play in the same way during
FIGHTING EBOLA WITH BLOOD
platelet transfusions. When we donate blood, it is often separated into three
component parts: red blood cells, platelets, and plasma.
Using genome-editing technology, scientists at ACT
Red blood cells are the oxygen-carrying component that
removed the gene required for the production of
bears the A, B, and O antigens we associate with blood-
the MHC proteins in platelets. As a result, these
typing. Platelets assist in blood clotting, and plasma is
universal platelets are not rejected by any recipient;
everything else—a pale, yellow liquid that makes up
however, it is a problem of special relevance to those
about 55% of the body’s total blood volume.
who receive platelet transfusions on a regular basis,
such as cancer patients undergoing chemotherapy or Plasma is about 95% water but also contains dissolved
radiation therapy. proteins, glucose, clotting factors, electrolytes,

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hormones, and antibodies: the proteins produced by is the plasma from a recovered patient contains
our immune system to fight infectious diseases. In the antibodies to Ebola. Rapid recognition of the virus by
current Ebola outbreak, plasma from individuals who the infused antibodies will alert an infected patient’s
survived the viral infection are used to treat freshly immune system, and hopefully put the virus out
exposed patients via transfusion. The presumption of commission.

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SCULPTING A BETTER LIFE FOR
SICKLE CELL PATIENTS Life Science Training from Industry Experts

With the recent announcement of two new biologics in fetal hemoglobin and a decrease in RBC stickiness.
in Phase I studies for Sickle Cell Disease (SCD), WEEKLY Although hydroxyurea does not work for all patients,
takes a closer look at the treatments for this autosomal it is turning back the hemoglobin clock on select
recessive disease, afflicting approximately 100,000 individuals for a better quality of life.
Americans and 25 million globally.
Monoclonal Antibody
Sickle cell anemia is caused by an SNP in the beta
Earlier this month, the FDA granted fast track
globin gene, which codes for hemoglobin—an oxygen-
designation to NKT Therapeutics’ (Waltham, MA)
binding protein in red blood cells (RBCs). The mutated
experimental drug for SCD, NKTT120. NKT Therapeutics
hemoglobin still transports oxygen, but this single base
derives its company name from the cells they target:
change (A to T) affects the protein’s shape in a way that
natural killer T-cells.
can have deadly consequences. Instead of existing as
individual proteins, the mutated hemoglobin proteins High levels of invariant killer T-cells (iNKT) are activated
clump together causing RBCs to form a “C” or “sickled” during an immune response in SCD. Repeated vaso-
shape. Sickled RBCs are unable to enter branch points occlusive events add increased inflammation to the
in the vascular system; they simply cannot fit through picture, which causes the iNKT cells to wrongly perceive
the narrowing blood vessels to feed the tissues of the its own tissues as foreign and attack accordingly.
body. Vital oxygen remains undelivered, potentially NKTT120 is a monoclonal antibody that targets and
causing the restriction of blood flow to an organ, known wipes out iNKT cells. Encouraging results from the
as a vaso-occlusive crisis, leading to severe pain and Phase I study are painting a prettier picture for better
damage. Individuals with two defective copies of the disease management.
beta globin gene have sickle cell disease. Those with
Gene Therapy
one defective beta globin gene have sickle cell trait.
bluebird bio (Cambridge, MA) is treating its first severe
There is no widely available cure for SCD at this time,
SCD patient with LentiGlobin BB305. This product is
aside from bone marrow transplants which can prove
derived from the patient’s own hematopoietic (bone
difficult due to the lack of closely related donors
marrow) stem cells, and altered via gene therapy to
that are often affected by the hereditary disease
produce a corrected beta globin gene. The corrected
themselves. Currently, most patients manage their
beta-T87Q-globin gene ultimately produces a non-
disease with preventive antibiotics, pain relievers,
sickling amino acid substitution which may prove to
and frequent blood transfusions. In this issue, we will
cure SCD. Initial clinical data on LentiGlobin BB305 is
discover how biotechnology may sculpt a better life for
expected next year.
sickle cell anemia patients.

A COLLECTION OF OPTIONS EASILY CONFUSED: ACUTE VS.

CHRONIC INFLAMMATION
Small Molecule Drug The word inflammation is derived from the Latin word
A common way to manage SCD today is a small “inflammo,” which means to ignite. Inflammation is
molecule drug called hydroxyurea, brand name Droxia the body’s attempt to remove harmful stimuli and
(Bristol-Myers Squibb, New York, NY). Hydroxyurea jump start the healing process. Signs of inflammation
turns on the production of fetal hemoglobin in adults include pain, heat, and swelling, lending itself to the
but the exact mechanism of action remains unknown. It Latin derivation.
appears hydroxyurea increases nitric oxide levels which Acute inflammation comes on suddenly and disappears
causes a cascade of reactions that ultimately starts when the outside stimuli is removed. It is associated
the production of gamma globin chains, necessary for
fetal hemoglobin production. Patients see an increase

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with infectious agents, the presence of foreign objects, SCD do not suffer from the symptoms during fetal
and burns. development or shortly after birth.
Chronic inflammation is prolonged inflammation
that can occur when the body itself activates the
THE TROPICAL ADVANTAGE:
inflammatory response, such as the vaso-occlusive
SICKLE CELL TRAIT
events of SCD. It is not easily resolved and therefore can Malaria may have shaped the propensity of SCD by the
last for significant time, inducing organ damage. way of sickle cell trait in certain tropical regions, such
as Sub-Saharan Africa. The malaria parasite spends
COCKTAIL FODDER part of its life in the RBC and causes the RBCs with
Before a baby is born, a slightly different form of defective hemoglobin to rupture prematurely, ensuring
the hemoglobin protein is produced, known as fetal the Plasmodium parasite is unable to reproduce. Those
hemoglobin. This hemoglobin has a higher affinity for with the sickle cell trait are still able to contract malaria,
oxygen. In other words, it binds oxygen more tightly. but their symptoms are generally less severe. In malaria
This tight oxygen binding prevents the hemoglobin from prone areas, chances of survival actually increase in
clumping together, and babies who will later develop people carrying the sickle cell trait.

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MUTATIONS AND DISEASE: THE SPICE OF LIFE Life Science Training from Industry Experts

This week’s issue features excerpts from the “Genetic if they touch a neighboring cell, they stop dividing.
Variation” chapter of The Biotech Primer, our 200-page Cancer cells have lost many of these controls or checks
book that provides an in-depth look at the biotech on cell division and start dividing and continue to
industry and the science that drives it. In this chapter, divide inappropriately.
we explain the different types and causes of genetic
mutations and then explore their relationship to THE GUARDIAN OF THE GENOME
disease and therapeutics. Today, we begin with a The p53 gene is the most frequently mutated gene in
discussion of genetic disease and end with a promising human cancer and the p53 protein it produces is often
drug target (53). called the “guardian of the genome”. Its role is to make
Just as Superman is drained of strength by kryptonite, a decision whether to repair DNA or kill the cell in
the Guardian of the Genome (53) loses its capacity response to DNA damage. To perform this function, p53
to fight disease when it is rendered powerless by has to bind to DNA in a very specific manner.
mutations. Just like Peter Quill (Star-Lord from Guardian Below on to the left is the structure of p53 bound to
of the Galaxy), the Guardian of the Genome does not DNA. Four domains of the protein bind to DNA in a
always get it right either. Mutations and disease: the cooperative manner. On the right is one of the domains
spice of life. bound to DNA where the DNA-binding surface of the
p53 molecule fits into the grooves of the DNA helix.
GENETIC BASIS OF DISEASE Most of the mutations that occur in human cancer—
Mutations play a large part in disease. In a monogenic mutation “hotspots”—occur in the DNA binding
disease, changes in one gene cause the disease. domains, close to the DNA-binding surface. Changes
Examples of monogenic diseases include sickle cell to domains can be enough to disrupt binding of p53 to
anemia, cystic fibrosis, and Huntington’s disease. DNA and disrupt p53’s guardian duties. Research into
In contrast, polygenic diseases are caused by the how protein structure abnormalities cause cancer and
interactions of many different genes. Polygenic diseases other diseases is leading to the discovery of new drugs,
are more common than monogenic diseases and some of which are designed to target the aberrant
include cancer, heart disease, Alzheimer’s disease, and protein structures.
Parkinson’s disease.
Polygenic diseases often have susceptibility genes
COCKTAIL FODDER
associated with them, which increase the likelihood Cystic fibrosis (CF) is an extreme exemplar of polygenic
of the person developing the disease, but do not disease. There are over 1,000 genetic mutations that are
absolutely predict its development. The ultimate known to cause CF. Vertex Pharmaceutical’s (Boston,
disease outcome will depend on other genes in the MA) Kalydeco (approved in 2012) targets a mutation that
individual’s genome, as well as environmental factors. impacts only 4% of all CF patients.
An example of susceptibility genes are the association
of breast cancer with the BRCA 1 and BRCA 2 genes. P53 A TRICKY DRUG TARGET
Any protein defective in more than 50% of cancers
POLYGENETIC DISEASE: CANCER is a tantalizing drug target; however, targeting p53
Cancer is described as uncontrolled cell growth. has proven to be tricky. Using today’s technology, it
Healthy cells have tight controls on cell division and is much easier to inhibit an overactive protein than
only divide in response to outside signals promoting to activate a defective protein. One possibility for
cellular growth and division. Non-cancerous cells also patients with mutated p53 gene sequences is gene
respond to signals that tell them to stop dividing. for therapy—delivering a correct copy of the gene.
example, most exhibit contact inhibition, meaning Gendicine (Shenzhen SiBiono GeneTech) is one such

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therapy approved by the Chinese State Food and Drug of a second protein, MDM2, to p53. Drugs that inhibit
Administration in 2003. The FDA rejected a biologics or prevent this MDM2-p53 interaction could potentially
licensing application for a similar therapy in 2008. result in restoring full activity to p53, enabling it to
Although many p53-associated cancers are caused by carry out its cell-protecting mission. Amgen, Roche,
mutations to the p53 gene itself, about half are the and Sanofi are among the companies pursuing MDM2
result of p53 inactivation caused by the tight binding inhibition therapies.

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SQUEEZING THE JUICE OUT
OF DRUG METABOLISM Life Science Training from Industry Experts

When popping a pill, we seldom think about what GFJ inhibit these transporter proteins, which results
happens next—to the pill, or even to our bodies. We in lower intercellular drug concentrations. Lower drug
assume the body welcomes any extra help to fix the concentrations correspond directly with loss of drug
problem, but the reaction is quite contrary. effectiveness. Continue to enjoy what remains a healthy
A swallowed pill (small molecule drug) is instantly treat, but just be sure to double check your medication
labeled by our body as foreign and the reaction of labels against the following examples:
our body is to immediately get rid of the unwelcome • Scientists have known for some time that statins
intruder. The process where the body breaks apart a such as Zocor (Merck) and Lipitor (Pfizer) can result
drug in order to rid itself of the foreign chemicals is the in increased levels via CYP3A4 inhibition with GFJ.
basis of drug metabolism. Understanding this process is • More recent studies indicate GFJ consumption may
a critical part of drug development. create toxicity issues with medications prescribed
Since the liver is the primary site where medicine to treat high blood pressure, such as Adalat
is metabolized, let’s focus on the liver enzymes (Bayer Pharma).
responsible for metabolizing small molecule drugs and • Drinking GFJ can cause escalated medication
squeeze out some insight as to how these enzymes levels in Zoloft (Pfizer) and BuSpar (Bristol-
impact drug safety and efficacy. Myers Squibb), both used as treatments for either
depression or anxiety.
A GLASS OF DRUG INTERACTIONS
• Erectile dysfunction patients should watch out
Ever notice warning labels on certain prescription
when consuming GFJ because Viagra (Pfizer) and
medications advising the patient to avoid grapefruit
Cialis (Eli Lily) have potential to become toxic.
juice (GFJ)? Although considered a healthy food, GFJ
contains certain chemicals that can inhibit key proteins • Allergy medicines such as Benadryl ( Johnson &
involved in drug metabolism. Depending on which Johnson) and Allegra (Sanofi) potentially lose their
protein is inhibited, the result will either be an increase effectiveness when consumed in tandem with GFJ.
in the blood concentration of the drug (to potentially
toxic levels) or a decrease in the amount of medication
TYBOST: MAKING A PATHWAY
reaching target tissues.
OUT OF A BYWAY
Inhibition of CYP3A4 is not always a bad thing. Last
Scenario 1: Increase in toxic levels of the drug.
week, the FDA allowed Gilead Pharmaceuticals’
CYP3A4, found in both the small intestine and the (Foster City, CA) Tybost to fly solo, a drug whose
liver, is necessary for the metabolism of certain primary function is to inhibit the cytochrome P4503A
drugs. GFJ inhibits the activity of CYP3A4, resulting enzymes. Tybost was previously approved in 2012 as a
in a superabundant accumulation of the drug in the component of a four-drug-combination anti-retroviral
patient’s bloodstream. These higher concentrations therapy used to treat HIV.
may cause direct toxicity or liver damage over time.
What is so special about Tybost? Some anti-viral drugs
High drug amounts force the liver to work harder to
are normally broken down by cytochrome P4503A
metabolize the drug. Only medicines broken down by
enzymes. When taken in conjunction with Tybost,
CYP3A4 enzymes are potentially affected by GFJ.
the combination results in boosted concentrations of
Scenario 2: Decrease in drug levels that reach the the antiviral medication for a longer period of time,
target tissues. resulting in greater efficacy.
Some medications use transporter proteins—proteins
on cellular surfaces that allow molecules to enter cells.

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EASILY CONFUSED: PK VS. PD TERM OF THE WEEK: PRODRUG
Pharmacokinetics (PK) is how the body affects a drug Some drugs are designed to be prodrugs—a drug
and Pharmacodynamics(PD) is how a drug affects given to patients in an inactive or less than fully active
the body. form—which is converted to the active form through
A necessary set of studies used during drug metabolism. A good example of an accidentally
development to access Pharmacokinetics is discovered prodrug is the anti-clotting medication
called AMDE: Plavix (Bristol-Myers Squibb, New York, NY).

• Absorption: the process of a drug entering the Initially, Plavix faced a recall due to an elevated rate of
blood circulation. ineffectiveness during treatment. As many as 14% of
patients on the medication still experienced strokes or
• Distribution: the dispersion of the drug throughout
heart attacks due to blood clots. Further investigation
the body.
revealed patients who had a mutation in the liver
• Metabolization: the body’s recognition and enzyme CYP2C19 were unable to activate the drug, thus
subsequent breakdown of the parent compound rendering Plavix ineffective.
into daughter metabolites.
Plavix’s continued access to the marketplace hinges
• Excretion: the removal of the drug from the body. upon prospective patients taking a genetic test to
determine whether they have the CYP2C19 mutation—
the one that makes them unable to activate Plavix.
Patients identified with the mutation are prescribed an
alternative medicine that uses a different mechanism
of action.

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TAILORING STEM CELLS TO FASHION
REPLACEMENT ORGANS Life Science Training from Industry Experts

If stem cells have their way, replacement organs may cord blood product indicated for disorders related to
find their place as a plentiful standard of treatment. the production of blood in the body. Clinical trials are
This pairing prompted us to wonder: what is it ongoing for stem cell-derived therapies for diabetes,
about stem cells that make them attractive to organ stroke, ALS, and spinal cord injury.
replacement therapies?
The therapeutic potential of stem cells lies in their
MIRACLE MESH WITH A
ability to differentiate into any cell type in the adult
SIDE OF STEM CELL
human body, depending on the type of stem cell. The Artificial Pancreas Project, spearheaded by JDRF,
However, a potential disadvantage of tissues or organs aimed to fund the development of new technologies
derived from stem cells is rejection by the recipient’s in diabetes. The goal was to replace the diabetic’s
immune system. A type of stem cell called an induced damaged pancreas and reduce or even eliminate the
pluripotent stem cell has the potential to turn down need for constant self-monitoring of blood glucose and
the need for the immunosuppressive therapy that goes insulin levels. The most exciting outcome after a decade
hand in hand with organ and tissue transplants. of labor? Viacyte’s (San Diego, CA) artificial pancreas
derived from embryonic stem cells, which gained
With these new methodologies premiering on the
approval to enter clinical trials earlier this month.
runway, let’s take a look at stem cells as they set trends
in tissue transplantation and organ replacement. Viacyte’s true innovation comes from their planned
delivery of these cells: encapsulation in a biocompatible
STEM CELL PRIMER mesh. This miracle mesh simultaneously shields the
The challenge for scientists working in the stem cell cells from the patient’s immune system, yet still allows
lab is figuring out the exact cocktail of growth factors, oxygen and nutrients to enter, supporting the cells’
hormones, and nutrients required to lead a stem cell growth and maintenance. Most importantly, insulin and
down the correct and intended developmental path. other hormones secreted by the pancreas are able to
To become a pancreatic cell, instead of a heart cell or a exit the device, potentially doing away with the need for
liver cell would be an example. Once the combination insulin injections.
is perfected, scientists can differentiate cell types What other organs are being derived from a
into their choosing. The ultimate goal is to use these combination of stem cells and device?
new cells to produce replacement tissue and organs Over the past few years, a handful of patients with
for patients suffering from degenerative diseases or damaged tracheas have received synthetic versions
traumatic injuries. attained from stem cells layered onto a fabricated
Embryonic stem cells are prized because of their scaffold engineered by Harvard Apparatus
potential to develop into any tissue in an adult Regenerative Technology (Holliston, MA).
human—a characteristic called pluripotency. By Tengion (Winston-Salem, NC) is currently conducting
reactivating four genes that have been turned off clinical trials using muscle stem cells overlaid on
during the progression from embryonic stem cell to a scaffold system to grow custom urinary conduit
specific tissue type, researchers can turn back the systems, used in patients who have cancerous
hands of time and create induced pluripotent stem cells bladders removed.
(IPSC). The IPSC advantage is that there is less chance of
Other structurally simple organs ideally suited for
rejection by a patient’s body.
scaffolding technology include esophagus, bladder, and
To date, the only stem cell-based therapy approved heart valves.
by the FDA is New York Blood Center’s Hemacord, a

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COCKTAIL FODDER BIOINKED: THE NEW REALITY SHOW
• The only place one may buy an organ legally is in Bioprinting is another approach for tissue production.
the country of Iran; however, citizenship is required Bioink, a mixture of cells (including stem cells) and
in order to lessen transplant tourism. hydrogel can be layered into three-dimensional shapes
• Australia is entertaining the idea of legal monetary using modified 3D printers.
compensation for organs. The first company to whip up a working bioprinter
• Singapore has yet to actually execute the is Organovo (San Diego, CA) with NovoGen MMX
process of paying donors but has legalized a Bioprinter. Since bioprinting its first blood vessel in
compensation plan. 2010, Organovo has jumped to presenting bioprinted
liver tissue for drug discovery earlier this year.
The bioprinting of skin (the largest organ in the human
body) onto burn wounds is being tested in clinical trials
sponsored by Wake Forest School of Medicine and the
Armed Forces Institute of Regenerative Medicine (Ft.
Detrick, MD).
At this time, complex organs cannot be produced due to
limitations in bioprinting and the challenge of creating a
robust vascular system.

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CAN CELL DEATH KEEP CANCER
PATIENTS ALIVE? Life Science Training from Industry Experts

Programmed cell death is making a name for itself ON THE MARKET: PROTEASOME
as a pathway in cancer drug development. We asked INHIBITOR DRUGS
ourselves: what is this process known as apoptosis and
As proteins in cells carryout their functions, they
why is it a sought after drug target?
naturally begin to suffer damage and break down. A
Apoptosis is a system that evolved in cells for detecting proteasome is a specialized compartment within the cell
damage to DNA and proteins within a cell. As a cell that is responsible for getting rid of damaged proteins
develops, apoptosis can kick in and direct the cell down via dismantling the peptide bonds that hold it together.
a self-destructive path, killing itself for the greater good
One way to manually activate apoptosis is by blocking
of the organism. It is the accumulation of damage within
or lessening the number of proteasomes. By stopping
the cell that triggers cell suicide, thereby removing
the proteasome from performing its function, the
cells from tissues before they affect nearby cells or
damaged proteins build up within the cell. Apoptosis is
become cancerous.
then activated and the cell terminates itself in response
The drug discovery industry is interested in learning to the accumulation of damaged proteins.
more about apoptosis as a means to better understand
What small molecule drugs use proteasome inhibition?
how some cancers develop and how to treat them. In
fact, some cancers are tied to defects in this suicide • Velcade, marketed by Millennium Pharmaceuticals
program—cells marked for death can survive and (Cambridge, MA) has been approved for the
continue to mutate long after they should have self- treatment of multiple myeloma.
destructed. Let’s take a magnifying glass to the process • Krypolis, developed by Onyx Pharmaceuticals
of apoptosis to see exactly what is has to offer in cancer (South San Francisco, CA) has been approved as a
drug development. second-line treatment for multiple myeloma.

EASILY CONFUSED: TYPES COCKTAIL FODDER: FACTS

OF CELL DEATH ABOUT APOPTOSIS
Apoptosis is an important part of cellular development • During the development of the nervous system,
and defense. A set program of biochemical changes neurons in the brain are selectively pruned to give
occurs after apoptosis is induced, including the rise to the final structure, thanks to apoptosis.
breakdown of the genetic material and the breaking
• The webbing between the fingers and toes of
away of small bits of the cell into separate membrane-
an embryo disappear during development due
bound vesicles known as apoptotic bodies. These are
to apoptosis.
recognized and destroyed by white blood cells.
• In the average human adult, between 50 and 70
In contrast, necrosis is cell death that is the result of
billion cells die each day and in children (8-14 years
an external injury, such as an infection or trauma.
old), between 20 and 30 billion cells dies per day
Unlike the apoptotic bodies produced during apoptosis,
because of apoptosis.
necrotic cells are usually not destroyed by white blood
cells. The necrotic process may result in the buildup
IN DEVELOPMENT: ARGININE-
of decomposing dead tissue and cell debris, further
DEPRIVATION THERAPY
resulting in gangrene.
Arginine-deprivation therapy is based on the fact that
certain types of cancer cells lack the ability to produce
the amino acid arginine—a necessary building block of
proteins healthy cells make for themselves.

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Areas targeted by arginine-deprivation therapy are the cancer cells are exploited here, potentially creating
stripped of their arginine, both healthy cells and a therapy less toxic than traditional cancer treatments.
cancerous cells. The healthy cells will replenish their Polaris Pharmaceuticals (San Diego, CA) is taking
lost arginine while the cancer cells will be unable advantage arginine-deprivation therapy with an
to replenish their arginine. This creates a state of experimental small molecule drug called ADI-PEG 20.
apoptosis due to the resulting damage from the loss of ADI-PEG 20 is in Phase III clinical testing for liver cancer
arginine, inclining the cancer cells to travel down the and Phase II testing for melanoma, mesothelioma, small
path of programmed cell death. The bonus is that only cell lung cancer, acute myeloid leukemia, and non-
Hodgkin’s lymphoma.

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MATTERS OF THE HEART Life Science Training from Industry Experts

In a picture perfect game of drug development, Angiotensin II is a small hormone. When it attaches
Novartis heartily delivered this week with LCZ696. to the angiotensin receptor, it causes a cascade of
Also propelling the world of cardiovascular disease reactions to occur. That cascade ultimately causes blood
treatments into high gear is a promising cholesterol vessels to constrict, which causes blood pressure to
therapeutic by Sanofi and Regeneron called alirocumab. increase. Think about it like this: it takes more pressure
Last week, Novartis released a heart failure drug to move fluid through a narrow tube than through a
called LCZ696, along with the data behind their highly wide one. Angiotensin II also promotes the release
successful Phase III trial. Novartis is hopeful, with sights of a second hormone, aldosterone, which increases
set on an FDA approval by year’s end. sodium retention by the kidney and further drives up
blood pressure.
Sanofi and Regeneron presented results that
support further development of a whole new class of By blocking the angiotensin receptor, angiotensin II has
therapeutics for high cholesterol, thanks to alirocumab. nowhere to land, no reaction cascade, no blood vessel
constriction and blood pressure is lowered.
It is a double dose of good news for hearts everywhere.
Let’s examine the science behind these new angles of Sacubitril is a neprilysin inhibitor; it stops the enzyme
attack against cardiovascular disease, which is currently neprilysin. Why is stopping neprilysin important?
the leading cause of death in the world. Neprilysin breaks down a hormone called natriuretic
peptide. By stopping neprilysin, we increase levels
TERM OF THE WEEK: HEART FAILURE of natriuretic peptide. Natriuretic peptide gets rid of
Heart failure, also known as chronic heart failure or sodium and dilates blood vessels; therefore, blood
congestive heart failure, is the inability of an individual’s pressure is lowered.
heart to pump and/or fill with blood adequate to the By combining an angiotensin II receptor inhibitor and
demands of the body. The causes of heart failure are a neprilysin inhibitor, the effect is enhanced, allowing
complex and varied but risk factors can include high LCZ696 to get to the heart of the matter.
blood pressure, diabetes, coronary artery disease—the
narrowing of blood vessels brought on by cholesterol- EASILY CONFUSED: HDL AND
containing plaque—and previous damage caused by LDL CHOLESTEROL
heart attacks. Lipoproteins are divided into two categories for the
sake of profiling cholesterol levels: high-density
BEATING HEART FAILURE lipoproteins (HDL) and low-density lipoproteins (LDL). A
An independent review panel ended the Phase III trial lipoprotein is a protein with fat molecules attached for
of Novartis’ heart failure drug LCZ696 last spring. The use as a transport system.
reasoning was that the experimental drug performs so
HDL is called good cholesterol because it transports
much better than the standard of care it is unethical to
the lipid component in a compact fashion without
continue testing. How does this wonder drug work?
losing or dropping the fat molecule when traveling in
LCZ696 is a combination drug: Valsartan and Sacubitril. the arteries. HDL can even scoop up and expunge the
They work together to lower blood pressure, which LDL, or bad cholesterol. LDL is more fluffy and detaches
lowers the strain on the heart and lessens the easily, wreaking havoc by oxidizing or subsequently
accumulation of fluid in the tissues, such as the attaching to arterial walls.
lungs—a key symptom of heart failure.
Valsartan is an angiotensin II receptor inhibitor; it stops BEATING BAD CHOLESTEROL
angiotensin II. Why is stopping angiotensin II important? Creating a buzz around a new pathway for fighting
bad cholesterol is Sanofi/Regeneron’s alirocumab and

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its strong clinical trials data. What is the story? First, a at the cell surface, impeding the process of removing
little background. LDL and breaking down cholesterol within the cell.
Low-density lipoprotein receptors (LDLR) are found on Alirocumab, a monoclonal antibody, binds to and
various cell surfaces, like liver cells, and they help the inhibits PCSK9. In a way, it protects the low-density
body get rid of excess cholesterol. The LDL particles lipoprotein receptor so it can uptake LDL without
bind to the low-density lipoprotein receptor, and the interference. It helps the LDLR to continue the cell’s
LDL is subsequently taken up by the liver cell. The LDL is process of breaking down cholesterol without the
broken down inside the cell and cholesterol is released threat of PCSK9.
to be used within the cell. The receptor, or LDLR, is More data is needed to clearly demonstrate the long-
recycled back to the cell surface, where it can bind to term safety and efficacy data of alirocumab, but these
and remove more LDL. results have generated much excitement as a validation
We also have another player, PCSK9, potentially of PCSK9 as a cardiovascular drug target. Preliminary
interrupting this process. PCSK9 binds to the low- results suggest that alirocumab cuts heart attack and
density lipoprotein receptor and both are taken in stroke rates in half.
by the liver cell; however, PCSK9 and the low-density Look for Merck and Amgen to jump on the bandwagon
lipoprotein receptor are degraded—sort of like a because both companies are also working on the
murder-suicide. This results in fewer receptors present development of monoclonal antibody inhibitors
of PCSK9.

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BREAKING THE ICE WITH ALS Life Science Training from Industry Experts

ALS awareness is on the rise, thanks to the ubiquitous suggests ALS is not actually associated with free
ice bucket challenges making worldwide headlines. This radicals, leaving the exact role of SOD1 mutations
social media phenomena prompts us here at WEEKLY unanswered. In some cases of sporadic ALS, incorrectly
to wonder: what is this disease and where does the folded versions of the SOD1 protein have been
biotech industry stand? detected, even though there is no genetic mutation.
ALS stands for Amyotrophic Lateral Scelerosis. Let’s
break it down to its roots:
TREATMENT PROFILE
Rilutek, marketed by Sanofi (Paris, France) is the only
• ’A’ means no
FDA approved therapy for ALS. Rilutek is thought to
• ’Myo’ means muscle work by acting as a glutamate inhibitor, although the
• ’Trophic’ means nourishment exact mechanism of action is not known.
“No muscle nourishment” means just that: motor A buildup of a neurotransmitter—a chemical produced
neurons tasked with sending impulses to the muscles by the body that transmits messages between
cannot do their job because they are degenerating. nerve cells—called glutamate has been observed
“Lateral” means side, indicating where the motor in the early stages of ALS. Glutamate is involved in
neurons are located within the spinal column. activating motor neurons. One theory is the excessive
“Scelerosis” refers to the scarring that occurs in the glutamate somehow damages the motor neurons by
region of the degenerating neurons. Because ALS overstimulating them. Rilutak potentially blocks the
affects the motor neurons (or neurons that control glutamate. While it is not a cure for ALS, it may extend
muscle movement), disease symptoms start with life by approximately sixty 60 to 90 days and possibly
muscle weakness and progress to paralysis, eventually slow overall disease progression.
eroding the patient’s ability to walk, chew, swallow,
speak, and in advanced stages, breathe. COCKTAIL FODDER: FAMOUS
Thanks to this viral campaign, the ALS Association has
FACES OF ALS
raised more than $100 million and recruited more than Prior to last month’s ALS campaign, the disease was
two million new donors to date, setting a precedent for commonly known as Lou Gehrig’s disease, referring to
rare disease fundraising efforts. Now that the ice cubes the Yankee’s baseball player. Lou Gehrig died in 1941
have started to melt, we will take a look at the science at age 37 after being diagnosed with the disease on his
behind the awareness. 36th birthday. The disease normally progresses very
rapidly (Lou Gehrig died almost two years after being
AN ELUSIVE ORIGIN diagnosed). On the opposite end of the spectrum is
Approximately 90% of ALS cases are of unknown 72-year-old physicist Stephen Hawking, a living example
origin. The other 10% are associated with FALS, or of how the disease can progress very slowly. Diagnosed
Familial Amyotrophic Lateral Sclerosis. Drug discovery at age 21, Hawking’s specific case illustrates how mental
is difficult for diseases whose mechanisms are not well function is typically not impacted by ALS.
understood or unknown. ALS is complex with many
factors potentially contributing to the demise of various
TREATMENTS IN DEVELOPMENT
organ systems via motor neuron death. There is plenty of interest in developing ALS treatments
with Biogen Idec (Cambridge, MA) announcing an
The most prominently identifiable culprit is one of
“ALS Innovation Hub” last month—an effort to better
several different mutations in a gene located on
understand the disease mechanism with the goal of
chromosome 21, known as SOD1. It encodes for
identifying new drug targets. And Prize4Life (Berkeley,
superoxide dismutase, an antioxidant enzyme that
CA) is inviting academic and industry contenders to
helps cells get rid of excessive free radicals. Research

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compete in finding a treatment for ALS. One million into ALS patients—where the cells differentiate into
dollars will begiven to the winner. Founded in 2006, motor neuron cells, essentially repairing the damaged
Prize4Life’s goal is to “accelerate the discovery nerves that are the source of the disease. Phase II
of treatments and a cure for ALS.” Registration is results have not yet been announced, but stem cell
still open. Could we have a winner from one of the treatment showed promising results in an animal model
treatments listed below? of the disease.
ISIS Pharmaceuticals (Carlsbad, CA) announced positive GlaxoSmithKline is in Phase II trials with monoclonal
results from a Phase I study of an antisense drug antibody Ozanezumab, aimed to maintain existing
targeting the SOD1 gene last year. The drug is aimed at muscle function in ALS patients with the possibility of
FALS and approximately 20% of FALS cases are tied to slowing the progression of the disease.
the mutated SOD1 gene. Lastly, ALS Therapy Development Institute (ALS TDI)
Neuralstem (Germantown, MD) is currently conducting (Cambridge, MA) screens over 25 different therapies
Phase II studies on a stem cell treatment for ALS. targeting ALS, check out their active pipeline.
Human spinal cord stem cells are surgically implanted

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CERDELGA: 25 YEARS IN THE MAKING Life Science Training from Industry Experts

We have a new orphan drug on the market thanks to has one copy of the gene but does not have disease
last week’s FDA approval of Cerdelga, a small-molecule symptoms can still pass that gene on to their children,
therapeutic developed by Sanofi for treatment of hence the term “carriers.”
Gaucher’s disease. This rare genetic disorder is thought • If the disease gene is dominant in both parents, all
to affect around 6,000 in the United States. Regular of their children will develop Gaucher’s.
WEEKLY readers know we like to highlight new drug
• If the disease gene is dominant in one parent and
approvals. After all, there are only 25 to 40 of them a
the other parent is a carrier, there is a 50% chance
year on average.
that their child will develop Gaucher’s and a 50%
Cerdelga is a drug 25 years in the making. What started chance the child will be a carrier.
as research at the University of Michigan in 1989 turned
• If the disease gene is dominant in one parent and
into a licensing and development deal with Genzyme
the other parent is unaffected, all of their children
in 2000 (now part of Sanofi). Over the last15 years, the
will be carriers and will not develop Gaucher’s.
drug was tested in clinical trials on over 400 patients in
29 countries, leading to the FDA’s approval and orphan • If both parents are carriers there is a 25% chance
drug distinction last week. that their child will develop Gaucher’s, a 50% chance
that their child will carry the gene recessively, and a
Gaucher’s is an inherited disease caused by a recessive
25% chance that they will be unaffected.
mutation in a gene located on chromosome 1. It leads to
the accumulation of glucocerebroside, a fatty substance • If one parent is a carrier and the other parent
essential in nerve cells, resulting in chronic fatigue and unaffected, there is a 50% chance that their child
easily fractured bones in the most common form of will be a carrier and a 50% chance that their child
the disease, type 1. To date, the standard of care has will be unaffected.
been regular infusions with recombinant versions of the
enzyme made in biomanufacturing facilities. SYMPTOMS EXPLAINED
The new oral treatment, Cerdelga, is a small How does lipid accumulation cause such serious
molecule inhibitor of the enzyme that produces the problems in Gaucher’s patients? Simply put, it interferes
glucocerebrosides in the first place—glucosylceramide with the normal functioning of the organs in which it
synthase. If the enzyme is not produced, it will not need accumulates. For example, lipid accumulation in bone
to be broken down. marrow interferes with the production of the platelet
cells required for successful blood clotting, leading to
Cerdelga is not the first oral treatment for Gaucher’s.
bleeding and bruising. The production of the oxygen-
Zavesca (Oxford GlycoSciences and Actelion) was
carrying red blood cells is also impacted, leading to
approved in 2003 as an oral treatment for use in
chronic fatigue and anemia.
patients who had only mild or moderate Gaucher’s,
or could not tolerate infusion therapy. With its
COCKTAIL FODDER: THE GAUCHER’S
better safety and efficacy profile, Cerdelga has been
DIABETES CONNECTION
approved as a first-line treatment—an idea that can be
easily swallowed. Zavesca, an earlier oral treatment for Gaucher’s,
was originally developed as an alpha-glucosidase
DOMINANT OR RECESSIVE? inhibitor when it was discovered that it also inhibited
glucosylceramide synthase. Today, alpha-glucosidase
We all have two copies of each gene—one that we
inhibitors are used for the treatment of type 2 diabetes,
inherited from our mother and the other from our
and work by preventing the digestion of carbohydrates,
father. In order to develop Gaucher’s and other
thus reducing their impact on blood sugar levels.
autosomal recessive disorders, one must inherit two
There is some preclinical evidence that a more specific
copies of the gene causing the disease. Someone who

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glucosylceramide synthase inhibitor, such as Cerdelga, therapy: provide them with the enzyme they need from
may also be effective at controlling blood glucose levels an outside source.
in type 2 diabetics. Substrate reduction therapy obviates the need for the
enzyme by reducing the amount of substrate—the
TERM OF THE WEEK: SUBSTRATE molecule upon which the enzyme acts.
REDUCTION THERAPY
For patients who do not generate enough of a needed
enzyme, the standard treatment is enzyme replacement

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A CHIP OFF THE NEW BLOCK Life Science Training from Industry Experts

PERSONAL GENOMICS TURNS potentially represent one gene (or gene variant) within
HEADS IN BIOTECH the human genome, even if the entire gene sequence
is not present. Typically between 25 and 60 bases are
23andMe, the Silicon Valley based personal genomics
used and high-powered software keeps track of the
company named for the 23 pairs of chromosomes
location of DNA sequences within the chip.
in a normal human cell, is turning a few heads in the
biotechnology industry lately. After a very public and A researcher starts the process by collecting the patient
still unresolved dust-up with the FDA’s decision to sample (typically saliva) through a mail-in DNA kit:
halt marketing of the company’s DNA-based personal • ISOLATE: extract DNA from the sample tissue.
“health reports,” 23andMe is looking for additional • AMPLIFY: make copies of the specific gene or DNA
ways to turn its vault of consumer DNA data into sequences to be examined using the polymerase
useful products in which the FDA will not interfere. chain reaction (PCR).
They recently announced a partnership with Pfizer to
• SORT AND LABEL: separate the sample DNA into
pinpoint the genetic causes of inflammatory bowel
single strands and label with a fluorescent probe.
disease (IDB). The hypothesis being tested: is 23andMe’s
consumer DNA data large and accurate enough to • LINE UP: wash the labeled sample DNA onto
characterize IBD disease states by a combination of the chip; The sample DNA sticks to (hybridizes)
genetic fingerprints rather than just symptoms? the DNA sequences on the chip if its bases
are complementary.
23andMe is the highest profile consumer genetics
company in the world and its foray into democratizing • CLEAN AND RECORD: wash off the unbound sample
genomic data gives rise to bold new avenues for DNA any sample DNA left is decoded.
human health—faster drug discovery, efficient patient • SOFTWARE SCAN: make correlations to a specific
screening, and improved preventative medicine. For all gene, or gene variant. Computer software keeps
these reasons, the technology behind 23andMe is well tabs on DNA positions and corresponding genes.
worth looking into.

A CHIP OFF THE NEW BLOCK

A tiny glass chip that is affixed with single strands of
DNA is what a DNA chip, gene chip, or SNP chip is. These
miniature transparent squares, called by many different
names, make up the core of 23andMe’s technology.
While it does give a global look into an entire genome,
gene chip technology is not the brute-force. Whole
genome sequencing (WGS) treatment is usually
reserved for simpler microbes and now humans,
thanks to Illumina’s next generation sequencing (NGS)
technology.
Gene chips take a look at our genome by finding the
patterns inherent in DNA. They rely upon the very A DNA chip or SNP chip that is spotted with different
specific base-pairing rules followed by all double- variants of a particular gene is used to specifically
stranded DNA molecules: A’s always pair with T’s; determine genetic derivatives associated with a
C’s always pair with G’s (adenine-thymine; cytosine- particular disease. DNA samples from patient groups,
guanine). Each single stranded DNA on a chip can such as IBD patients, are compared with DNA samples
from healthy individuals. The data is further analyzed

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to see if particular variants are more common in the Even though 23andMe made it clear to consumers
patient vs. healthy population and vice versa. that their test results were not diagnoses but rather
probabilities, the concern remained because those
FDA HITS PAUSE ON DTC probabilities are based on a changing scientific
GENETIC TESTING understanding of the vast unknowns.
The “direct to consumer” (DTC) genomics revolution that
23andMe purports to lead took a hit last year when the EASILY CONFUSED: SNP
FDA ordered them to stop selling their personal health ANALYSIS AND SEQUENCING
product to consumers. The term “DNA sequencing” is often misconstrued
The DNA chip technology behind their product is with the term “SNP analysis” during discussions. DNA
considered a reliable and accurate research tool by sequencing determines the order of every single base
industry standards. What happened? pair in a given gene (gene sequencing) or in an entire
genome (whole genome sequencing). SNP analysis
Well, it turns out that some SNP variants have a strong
identifies single base differences between a given DNA
enough statistical association to be used as a diagnostic
sequence and a reference DNA sequence.
(for example, the BRCA1 mutation); however, not all SNP
variants have this capacity, which in theory could lead to TERM OF THE WEEK: SINGLE
flocks of fazed consumers making faulty assumptions. NUCLEOTIDE POLYMORPHISM (SNP)
23andMe’s process involves SNP analysis on a cheek cell A single nucleotide polymorphism (SNP for short,
sample obtained from saliva provided by consumers pronounced like snip) is a one base difference in
through a mailed DNA kit. The analysis provides an the DNA sequence of a gene when compared to the
estimate of an individual’s statistical risk based on sequence found in the majority of the population.
a limited analysis of key genetic regions known to Many SNPs have no significant impact on an
contain mutations associated with disease—like type individual’s health, but others are associated with
2 diabetes, age-related macular degeneration, and disease susceptibility.
many more.

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CULTIVATING BIOLOGICS FROM PLANTS? Life Science Training from Industry Experts

A FRESH LOOK AT TOBACCO (or plant cells) keeps spending in check. Additionally,
AND BIOLOGICS plant production capacity can be quickly scaled to
meet demand. Trumping everything, producing human
Tobacco plants pioneering the production of
therapeutics in plants is considered to be the safest
monoclonal antibodies for an experimental Ebola
option since it is unlikely any human pathogen could
treatment turned out to be quite the head scratcher
infect a plant. Mammalian cell lines can and do become
for last Thursday’s WEEKLY readers. With tobacco’s
infected with human pathogens.
reputation polluting our recent memories, let’s take a
fresh look at tobacco and biologics. The most efficient way to turn plants into therapeutics
factories is via viral vectors. The process goes
To date, no approved therapeutic protein has been
something like this:
produced in whole plants—ever!
• First, insert therapeutic protein gene(s) into a
In 2012, Israeli company Protalix successfully earned
plant virus
FDA approval for its plant cell produced drug ELELYSO,
a recombinant version of the glucocerebrosidase (GCD) • Next, allow plant infection to occur
enzyme deficient in Gaucher’s disease patients. This • Then, wait for plant to produce viral proteins, which
nod from the FDA demonstrates that plant cells do now include the therapeutic proteins we are after
have the ability to produce complex, therapeutically • Lastly, harvest plants by extracting and purifying
functional human proteins. the therapeutic proteins
Taking a walk on the animal side, remember it is the Tobacco plants, oddly enough, are the most commonly
mammalian cell line—Chinese Hamster Ovary (CHO) used plant for therapeutic protein production.
cells—that are the cell of choice for production of most Cultivated by humans for centuries, researchers have
complex protein therapeutics, such as monoclonal studied one of the tobacco plant’s most common viral
antibodies. Biologics produced in plant cells are more afflictions—TMV, or tobacco mosaic virus -- and as a
cost effective than mammalian cells. Biologics produced result, it is one of the most well understood viruses.
in whole plants are an even thriftier solution. This plant, with its hand in countless premature deaths,
A few companies who are developing whole may prove a boon to public health as it may enable
plant production platforms for therapeutics and cheaper, faster production of life-saving therapeutics.
vaccines include:
• Planet Biotechnology (Hayward, CA) for dental
ACHOO!
caries (tooth decay) and common cold. The dreaded cold. Why is there no cure?
• Mapp Biopharmaceuticals (San Diego, CA) for Simply put, it is too expensive to produce a cure via
diarrhea (caused by Clostridium difficile), AIDS, and conventional biomanufacturing. However, tobacco
Respiratory Syncytial Virus (RSV) infection. plants may be the answer to our collective winter woe.
• University of Louisville, Kentucky Bioprocessing / The ICAM-1 is the gateway receptor in which the
Intrucept Biomedicine (Owensboro, KY) for AIDS. rhinovirus invades our respiratory tract cells. One
potential treatment is soluble receptors—engineered
THE ROOT OF PLANT versions of the ICAM-1 receptor. When these man-
PRODUCED THERAPEUTICS made ICAM soluble receptors are placed in the vicinity
Using genetically engineered mammalian cells to of the rhinovirus, they are able to attract and “mop
produce biologics—thousands of liters in carefully up” the cold virus before it can hook onto and enter
monitored production tanks—is a pricey proposition. respiratory cells.
Producing the same amount of drug in whole plants

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Since Planet Biotechnology’s tobacco plant platform Preliminary positive results have been achieved with
technology is able to produce ICAM soluble receptors, edible vaccines for cholera in bananas and HBV in
an inexpensive biomanufacturing option may help potatoes. The gut contains specialized immune cells
make the cure for a common cold a reality. capable of detecting specific antigens which are not
destroyed by the digestive process and then activating
TERM OF THE WEEK: other immune cells to target those antigens. This
BIOMANUFACTURING research is still mostly at the preclinical stage, but it
Biomanufacturing is the use of living cells to produce offers up the tantalizing possibility of engineering food
a product. Typically, the term refers to the use of large items to contain an immunogenic antigen.
volumes of genetically engineered cells, such as E. coli • Icon Genetics’ (Halle, Germany) tobacco plant
bacteria or mammalian cells such as Chinese Hamster produced non-Hodgkin’s lymphoma vaccine has
Ovary (CHO) cells, to produce a product. successfully completed Phase I clinical studies. The
Engineering plants to produce a therapeutic or vaccine gene, encoding the patient-specific tumor antigen,
is another type of biomanufacturing sometimes cleverly is used by the plants to produce the personalized
referred to as biopharming. vaccine. The expressed protein is extracted from
leaves and injected into the patient in order to
MARKET WATCH: GARDEN EDITION boost their immune response against cancer.

Researchers are cooking up the possibility of • Medicago (Quebec and Research Triangle Park, NC)
edible vaccines delivered via bioengineered fruits also tumbles into the mix with a plant-produced
and vegetables. influenza vaccine in Phase II clinical studies.

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TINY VIRUS, BIG PROBLEM Life Science Training from Industry Experts

YEARS IN THE MAKING, EBOLA MARKET WATCH:

BUBBLES TO THE SURFACE EBOLA THERAPEUTICS
Bubbling under the surface for years like a volcano Unlicensed antibody therapy Zmapp takes the path
packing heat, Ebola erupts into a stark reality for the less traveled when used to treat two Ebola patients on
world health community. US soil.
First appearring in 1976, Ebola is a classified “Category Mapp Biopharmaceuticals (San Diego, CA) developed
A” potential bioterrorism agent. With an incubation Zmapp while working on antibody cocktails to fight
period of up to 21 days (early symptoms mimick Ebola. The serum has yet to reach even a basic Phase I
the everyday flu), it is entirely possible that infected human trial—as is the case with all except one potential
individuals board planes, unknowingly carrying the Ebola therapeutic. The basic idea behind Zmapp is
virus around the world. Given the threat, developing for its antibodies to neutralize Ebola by binding to
vaccines and treatments is an urgent priority. the virus and preventing it from entering the target
The World Health Organization (WHO) has confirmed cells. Therapeutic antibodies are typically produced
932 deaths through August 6th, mostly in Liberia, Sierra in Chinese hamster ovary (CHO) cells, but Zmapp’s
Leone, and Guinea. In total over 1,700 cases have been antibodies are produced in tobacco plants.
reported. WHO has pledged $100 million to help bring An RNA interference (RNAi) drug developed by Tekmira
the current outbreak under control. Pharmaceuticals (Vancouver, British Columbia) is
In this issue, we will learn how Ebola causes its targeting Ebola by silencing its genes. The drug
destruction and the therapeutics and discuss vaccines demonstrated efficacy bycontrolling the virus in non-
in development for the virus. human primates and began human safety testing in
January of 2014. An elevated inflammatory response to
TINY VIRUS, BIG PROBLEM the drug caused the FDA to impose a clinical hold on the
TKM-Ebola Phase I study. Tekmira is currently preparing
Seven proteins, a long single strand of RNA, and a lipid
a Complete Response to the Agency and Tekmira hopes
membrane make up the intelligent, yet calamitous
to resume testing by Q4 2014.
Ebola virus. How does a handful of proteins and genetic
material wreak such havoc? White blood cells called Duplicating a drug used to treat other viruses (such
macrophages become infected during an integral part as herpes and hepatitis), BioCryst (Durham, North
of the immune process; Ebola disrupts the body’s Carolina) is developing an inhibitor to an enzyme that
defenses against foreign invaders. Ebola uses to make copies of its genetic material. Since
the inhibitor has a chemical structure similar to the
Macrophages normally release signaling molecules to
building blocks that make up RNA, Ebola unknowingly
alert and activate other white blood cells to help fight
incorporates these inhibitors into its growing RNA
infection. Simply put, Ebola inhibits macrophages from
strand. The secret—once incorporated the RNA strand
doing their job. The virus stealthily evades the immune
cannot grow any longer, thus stopping Ebola replication.
system and spreads to the lining of blood vessels before
BioCryst’s drug is still in preclinical development.
moving to liver and kidney tissue. Infected endothelial
cells (the cells that line blood vessels) become leaky,
COCKTAIL FODDER
giving rise to the gruesome hemorrhaging characteristic
of infection. Novartis’ blockbuster leukemia drug, Gleevec, just
might have crossover potential as an Ebola drug. Both
Gleevec and the related drug Tasigna inhibit an enzyme
called Bcr-Abl that is overactive in certain types of
leukemia. Ebola uses a related enzyme called cAbl1 to

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help its reproduction. Gleevec and Tasigna appear to • They can protect against multiple viral strains
inhibit viral replication of Ebola-infected cells in the lab at once.
via the enzyme pathway. • They are much more transport stable than
protein or virus-based vaccines, which takes acute
MARKET WATCH: EBOLA VACCINES importance considering their intended deployment
The premise behind vaccines is straightforward: in developing countries; they will be less dependent
induce an immune response by injecting the patient on refrigeration in order to maintain efficacy.
with a killed or weakened virus—both traditional
The development of DNA vaccines has been elusive
vaccine preparations.
due to inefficient delivery methods. However, Inovio
Ebola may necessitate a different strategy: a Pharmaceutical (Blue Bell, PA, and San Diego, CA)
DNA vaccine. has a DNA vaccine program that holds promise of
How do DNA vaccines work? Inject a viral gene (that demonstrated efficacy in preclinical mice testing
provides the recipe to make a particular viral protein) models. Inovio has developed an electroporation-
into a patient so that their own cells manufacture based delivery system: injection of a vaccine into skin
that viral protein. The viral protein is unable to cause or muscle followed up with short, controlled electrical
disease, but is able to elicit an immune response. That pulses which significantly increases the tissue’s abililty
response makes memory B-cells which act as sentries to take up the DNA vaccine. Perhaps this delivery
that watch out for particular pathogens to appear, and method could be employed for an Ebola DNA vaccine?
then quickly wipe them out. GlaxoSmithKline’s recent acquisition of Okairos, a
The advantages of DNA vaccines: Swiss biotech with a DNA-based Ebola vaccine in its
preclinical development portfolio, is also jumping on
• They likely induce a more robust immune response.
the Ebola bandwagon.

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PLAYING DETECTIVE WITH
CHECKPOINT THERAPIES Life Science Training from Industry Experts

PLAYING DETECTIVE WITH these immunotherapies. In a promising development,

Bristol Myers Squibb has developed a PD-1 inhibitor
CHECKPOINT THERAPIES
recently approved in Japan for the treatment of
Over the past two weeks, we have discussed cancer melanoma. The drug is marketed by Japanese partner
immunotherapies that involve training T-cells (the Ono Pharmaceutical under that brand name Opdivo.
detective warriors of our immune system) into seeking
In May, Roche won the FDA’s coveted breakthrough
and destroying cancer cells.
therapy designation for their PDL-1 program, potentially
This week, we will take a look into another approach putting it on an inside track at the agency, which has
that has been creating a lot of recent excitement: been hurrying along new medicines in the pipeline.
checkpoint inhibitor drugs. Simply put, investigators
believe they have figured out a way to dismantle the THE FLIP SIDE
mechanism used by cancer cells to stay hidden from
T-cells. Overriding immune system checkpoints could
potentially have some negative effects-–namely the
CTLA-4 is a protein on the surface of T-cells that acts as activation of autoimmune-like symptoms in patients.
an off switch for the sleuthing T-cells. It signals them to
remain in a resting state, kind of like Sherlock lacking his One reason researchers are so excited about PD-1
investigative instincts. Yervoy (Bristol Myers Squibb) is a inhibitor drugs is the clinical data that indicates fewer
monoclonal antibody that targets these CTLA-4 proteins adverse reactions than those encountered with CTLA-
and prevents them from sending the off switch signal, 4 inhibitor drugs. PD-1 inhibitors work by targeting a
which allows the T-cells to turn on their remarkable mechanism common to many cancers – “cloaking” their
ability to sniff out disguised cancer cells. sinister activities behind the PD-1 receptor. Many are
hopeful that PD-1 inhibitors will be broadly effective
Yervoy was the first checkpoint inhibitor drug approved against a range of cancers and have safer profiles.
by the FDA in 2011 and it is used to treat melanomas
that cannot be removed by surgery.
COCKTAIL FODDER
Two other notable checkpoint proteins are PD-1 and
Preclinical studies suggest that PD-1 inhibitors may also
PDL-1, the Dr. Watsons of this story. PD-1 is another
be effective in treating HIV infection. HIV uses infected
inhibitory protein on the surface of T-cells; its inhibitory
T-cells to disable the immune system. Human clinical
activity is turned on when it meets the PDL-1 protein
trials for the HIV indication are in planning stages.
on the surface of host cells. Some types of cancer cells
have increased amounts of PDL-1 on their surface
making them experts at evading the immune system.
TERM OF THE WEEK: IMMUNE
SYSTEM CHECKPOINT
Fear not, for a number of companies, including Bristol
Myers Squibb, Roche, and Merck are developing In order to prevent autoimmune disorders, our immune
inhibitor drugs for PD-1 and PDL-1. The treatment of a system has evolved “checkpoints”—proteins on immune
variety of cancers, including melanoma, kidney cancer, cells that need to be switched on or off in order to start
and non-small cell lung cancer could soon be less of a the appropriate immune response.
mystery and more of a reality. Some cancer cells have cleverly evolved ways to exploit
Read more: Immune therapy’s cancer promise creates these checkpoints and proliferate under the guise of
research rush (Bloomberg). mystery. Immune system checkpoint therapies thwart
the criminal cancer cells early on by taking control of
the switch before it has flipped.
ON THE CASE
While PD-1 or PDL-1 checkpoint inhibitor drugs have yet
to be approved by the FDA, there is momentum behind

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SCI FI SCIENCE: COOL IN MOVIES,
COOLER IN CLINIC Life Science Training from Industry Experts

PRIMED TO BEAT PROSTATE CANCER virus particles, which in turn infect and destroy the
remaining cancer cells. This further stimulates the
We usually associate vaccines with infectious disease—
patient’s own immune system to target the tumor.
measles, mumps, and polio. This 20th century miracle
of medicine is being rolled out again, but this time Talimogene laherparepvec or T-VEC is in Phase III
with a 21st century mission: target cancer. In this development by Amgen (Thousand Oaks, CA) for
week’s issue, we will continue our discussion of melanoma and head and neck cancers consisting of
immunotherapies by focusing on two cancer vaccine genetically engineered herpes virus. Why herpes? The
technologies that could be straight out of an Isaac engineered herpes virus no longer causes herpes,
Asimov book. targets and infects tumor cells, replicates only in
tumor cells, and produces the protein GM-CSF—an
Let’s begin with a cancer vaccine currently available in
immune-stimulating protein. If effective, this platform
the US: Provenge, marketed by Dendreon (Seattle, WA),
technology may also prove successful in a number of
primes a patient’s body to attack their prostate cancer.
other cancers.
Let’s take a closer look at this innovative approach to
targeting cancer. EASILY CONFUSED: PREVENTATIVE
The science driving the technology: white blood cells, VACCINE VS. THERAPEUTIC VACCINE
when treated with signaling proteins, can be coaxed Preventative vaccines target viruses linked to specific
into becoming dendritic cells. Dendritic cells are highly types of cancer that a person may become infected
specialized immune cells. Their job is to break down with in the future. The most significant example is the
foreign proteins into smaller pieces then display those human papilloma virus (HPV) vaccine, Gardasil (Merck).
smaller pieces on their own cell surface. When T-cells HPV infection is a known risk factor for cervical cancer.
see the displayed foreign protein fragments, that Vaccination against it should reduce the occurrence of
presentation prompts the T-cells to seek and destroy cervical cancer. Likewise, vaccinations against hepatitis
any cell to which that protein might be attached. B virus may prevent liver cancer.
The science driving the therapeutic Provenge: a Therapeutic vaccines, in contrast, attempt to get the
patient’s white blood cells are collected, treated, and immune system to launch an attack against a cancer
induced to become dendritic cells. These patient- that is already established.
specific dendritic cells are exposed to a protein,
prostatic acid phosphatase, which is overexpressed COCKTAIL FODDER
in prostate cancer. When infused back into patients,
The Bacillus Calmette-Guerin (BCG) vaccine can be
these treated dendritic cells—displaying prostate
used as either a preventative or therapeutic vaccine,
cancer-specific protein fragments—cue the patient’s
depending on the indication.
own T-cells to launch an attack against the prostate
cancer cells. Provenge is also called an “activated blood BCG is a somewhat unusual approach to
product” because this therapeutic vaccine activates the immunotherapy for treatment of some bladder cancers.
patient’s white blood cells to fight cancer. A strain of live, but weakened, cow tuberculosis
bacillus bacteria is injected into the patient’s bladder
SCI FI SCIENCE and the presence of the bacterium activates an
immune response. The BCG vaccine is effective against
An exciting therapeutic cancer vaccine still in
approximately 66% of superficial bladder tumors.
development is referred to as an oncolytic virus. An
oncolytic virus is one that infects and kills cancer BCG is better known as a preventative TB vaccine, a
cells. The science fiction part—when the cancer cell vaccine used since 1921.
is destroyed it burst open and releases new oncolytic

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TERM OF THE WEEK: ADJUVANT Preventative vaccine adjuvants stimulate the
production of B-cell memory cells so the body is primed
Both preventative and therapeutic vaccines are often
to attack future invaders.
delivered with an adjuvant: a substance to help boost
the body’s immune response. Therapeutic vaccine adjuvants stimulate the production
of T-cells so the body can attack established invaders.

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DISSOLUTION OF CANCER CLUB WANTED Life Science Training from Industry Experts

DISSOLUTION OF CANCER Today, scientists are engineering T-cells to recognize a

CLUB WANTED specific tumor antigen. T-cells are removed from the
patient’s blood. Researchers use viruses (stripped of
If you are part of the “cancer club,” it means that you are
their ability to cause illness but retain their capacity
personally familiar with the devastation cancer causes.
to integrate into cells’ DNA) to deliver genetic material
Far too many of us are members, whether it is through
to produce a modified T-cell receptor. This modified
supporting loved ones with cancer or enduring and
T-cell is called a chimeric antigen receptor T-cells (CART)
persevering with it ourselves.
which is able to recognize proteins on the surface of the
It has long been the goal of physicians and researchers patient’s tumor. CART cells are then multiplied in the lab
who specialize in cancer treatment to harness the and infused back into the patient’s blood, where they
built-in power of a patient’s own immune system to ferret out and destroy tumor cells.
fight this dreaded disease. The ability to successfully do
so means more effective and less toxic treatments for IN THE CLINIC: LIVING DRUGS
cancer patients. CART therapy originated from research done by
Monoclonal antibody (mAb) therapies that bind Dr. Carl June at the University of Pennsylvania,
to cancer cell-specific proteins and induce an and subsequently was licensed and developed by
immunological response were the first treatments that Novartis. Other companies pursuing CART include Juno
achieved the goal. Since the approval of the first mAb Therapeutics, Kite Pharma, Cellectis, and Celgene.
cancer therapeutics in the late 1990s, cancer-specific Although still in early-stage clinical trials, initial results
immunotherapy has come a long way. This week and offer much hope: in several leukemia patients, cancer
next, we will take a look at some of the hottest new cells were reduced to undetectable levels!
cancer-fighting immunotherapies in development.
One potentially troubling side effect was observed:
BUILDING BETTER WARRIORS in some patients, infusion of the newly engineered
T-cells induced a strong inflammatory reaction causing
Circulating throughout your body is an army of stealth
serious symptoms such as fever and precipitous drop
warriors. When they do their job correctly, you do not
in blood pressure. However, these symptoms were
even know they are there or appreciate the perils they
successfully managed with drugs already approved to
save you from. These warriors are known as T-cells—a
treat inflammatory conditions.
specific type of white blood cell that seeks and destroys
virus-infected cells and tumor cells. If engineered T-cells continue to live up to the hype,
we can look forward to what some have termed a
Each T-cell has a unique receptor protein on its surface
“living drug”—cancer-fighting cells that will persist and
that recognizes a specific target protein. As T-cells are
multiply in a patient’s body until the tumor is destroyed.
completing their development, receptors that recognize
self (proteins on the surface of the host’s own tissue) COCKTAIL FODDER
are destroyed to prevent autoimmune disorders. This
In non-humans, a few cancers are transmissible among
protection against autoimmunity, however, means
the same species; Sticker’s sarcomas in dogs and devil
some tumor cells inevitably escape detection since
facial tumor disease in Tasmanian devils.
tumor cells are simply one’s own cells dividing too
rapidly or too frequently. Often, it is not until later in the
cancer cells’ development that a tumor antigen appears,
TERM OF THE WEEK: IMMUNOTHERAPY
enabling it to be targeted by the immune system. Immunotherapy: therapies that harness the power of a
patient’s immune system to combat a disease.

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INHALING INSULIN IS A BREATH OF FRESH AIR Life Science Training from Industry Experts

INSULIN INHALED to the development of techniques used to develop

inhalable insulin.
Injections are a part of life for most diabetics. Type 1
patients inject insulin into themselves multiple times YOUR DELIVERY MAKES
per day, and as many as 40% of type 2 diabetics require ALL THE DIFFERENCE
at least one daily injection.
Inhalable insulin offers the obvious advantage of
Thanks to the FDA’s approval of MannKind Corporation’s enabling diabetics to forego at least some of their
inhalable insulin therapy called Afrezza, needles just needle sticks. In addition, the lungs offer tremendous
might be a thing of the past for some diabetics. potential for drug delivery. Tiny air sacs called alveoli
This announcement was a long time in the making. greatly increase the internal surface area of lungs,
Since starting late-stage clinical studies almost eight which allow inhaled drugs abundant and direct access
years ago, the company has spent $1.8 billion dollars, to arterial blood as opposed to just capillaries, enabling
conducted more than 60 clinical trials on 6,500 patients, a potentially shorter time to reach its target.
and forged on after two FDA rejections. In this week’s Let’s remember that Afrezza is not the first inhalable
newsletter, we will look at the potential benefits of insulin to be approved by the FDA. Back in 2006, Pfizer’s
inhalable insulin and examine how it is made. Exubra was the first, only to be withdrawn just a year
later due to poor sales. What happened and why should
FROM PLANTS TO PATIENTS MannKind hope for a better outcome?
Protein therapeutics on the market today are delivered
First: the delivery device. Exubra’s device was large
into the body invasively. They are injected into muscle,
and unwieldy, making it difficult to carry and use
under the skin, or directly into the bloodstream. Why
discreetly in public. In contrast, MannKind’s delivery
so harsh?
device is similar to a small asthma inhaler, making it
The digestive system, by nature, is terribly inhospitable highly portable.
place to be. If protein drugs were to be taken orally, the
Second: the potential medical benefits. When taken
patient’s digestive track would break them down in the
just before a meal, Afrezza appears to more closely
same way it digests proteins consumed in food. Proteins
mimic natural insulin spiking than Exubra or injectable
are very sensitive to their environment, which is why
insulins, with blood concentrations peeking around
biomanufacturing is such a critically monitored phase
14 minutes post-injection as opposed to 45 minutes.
of biologics development. The wrong formulation of a
Clinical trials data also suggests that patients
properly manufactured protein can kill its efficacy. How,
receiving Afrezza have fewer incidences of insulin-
then, is inhalable insulin possible?
induced hypoglycemia.
Inhalable insulin traces its roots back to work done
Both of these points suggest that Afrezza has a better
by plant physiologist Carl Leopold in the early 1990s.
chance of success than the ill-fated Exubra.
Leopold observed that seeds from certain plants
such as soybeans with high protein content could COCKTAIL FODDER
become very dehydrated, yet be completely revived
MannKind’s founder, Alfred Mann, was also the inventor
when they reabsorbed water. The functionality of
of the insulin pump.
the plant proteins had been preserved, even in the
dried out state. Further analysis showed that survival
ability depended on the sugar content of the seeds.
ON THE HORIZON
Leopold used these observations as the basis for If successful, MannKind’s proprietary Technosphere
developing methods to preserve small proteins such particle used to deliver insulin may soon be in demand
as insulin in powder form. This work eventually led for the delivery of other biologics. This is no small feat

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and even if inhalable insulin is successful, adapting this Even more revolutionary than inhalable insulin would
technology to other proteins is not a sure bet. At only be oral insulin. Toronto-based Generex has an oral
51 amino acids, insulin is among the smallest proteins formulation for insulin in Phase III trials. Their RapidMist
around, and has a correspondingly simple structure. inhaler delivers insulin particles that have been
Larger, more complex proteins may not be as amenable formulated to be easily absorbed through the mucous
to inhalation. membranes of a patient’s mouth, thereby avoiding the
digestive system.

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BIOTECH CAKE AT YOUR NEXT GLUTEN-
FREE BIRTHDAY PARTY? Life Science Training from Industry Experts

NO BREAD FOR YOU: CELIAC DISEASE BIOTECH CAKE AT YOUR

Imagine walking into a bakery and smelling the NEXT BIRTHDAY PARTY
tantalizing scent of freshly baked bread but instinctively Imagine you are ten years old with celiac or gluten
dread the extreme discomfort indulging in a piece will sensitivity. Pizza, cupcakes, cookies, pretzels -- sadly, all
bring. This is the reality for the millions of Americans of these are off limits. While your friends are biting into
diagnosed with celiac disease. tasty treats, you clutch the wrapper of your gluten-free
Celiac disease is an autoimmune disorder triggered by standby snack your mom shoved in your hand as you
the ingestion of gluten proteins from wheat. Typically, cross the threshold of envy: birthday parties—the bane
the proteins in our diet are broken down into their of your breadless existence.
chemical building blocks (amino acids), which the As we said earlier, there is currently no treatment for
intestines then harmlessly absorb in order for our cells celiac and gluten-sensitive patients aside from going
to make new proteins essential to cellular function. gluten-free. Learning specialized ingredients names
Gluten proteins, however, have unusually high levels disguised as gluten is not much joy for a ten year old.
of the amino acid proline, which makes them resistant This is difficult to do as wheat and wheat derivatives
to digestive enzymes. This incomplete digestion are common ingredients across a wide range of foods,
results in the production of amino acid chains called tricking even the most discerning eye.
peptides. In susceptible individuals, these peptides Of course, gluten can be extracted from wheat but
stimulate an immune response directed towards their since it contributes to the elasticity of dough (aiding its
own intestines. ability to rise), gluten-free bread recipes often lack that
Symptoms of celiac disease can range from diarrhea, springy texture that folks miss, often resembling more
weight loss, and malnutrition to less obvious symptoms of a cardboard-like texture.
such as isolated nutrient deficiencies, but no However, biotech just might have an answer for the
gastrointestinal complaints. more than twenty million Americans who suffer from
Currently, the only treatment for celiac disease either celiac disease and gluten-sensitivity.
sufferers is avoidance of foods containing gluten. Wheat glutens include two different proteins: gliadins
and glutenins. It is the gliadin proteins that give rise
TERM OF THE WEEK: PEPTIDE to the problematic peptides and provoke an immune
A peptide is defined as a chain of amino acids consisting response. Using genetic engineering technology, a
of 50 or fewer amino acids. group of researchers from the Spanish Institute for
Sustainable Agriculture have developed a strain of
Peptides may be produced as part of the digestive
wheat lacking the gliadin proteins. In the group’s most
process of proteins, but there are also several examples
recent paper in the journal Public Library of Science
of peptides produced by the body to serve a biological
One, they tested the gliadin-deficient wheat in the
function. For example, the peptide hormone glucagon
production of bread. They found it showed in baking
is produced by the pancreas in response to low blood
that the sensory and overall acceptance was similar
sugar. Glucagon signals the liver to release stored
to that of normal flour as evaluated by a panel of
glucose into the bloodstream. Other biologically
trained assessors. This is likely due to the fact that the
important peptides include amylin (slows stomach
non-allergenic glutenin proteins were still present in
emptying) and prolactin (stimulates female mammals to
the wheat.
produce milk).
Additional studies, such as feeding trials with gluten-
intolerant patients, are still needed before this type of

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bread will be available commercially, but its a notable blood cells that attack foreign invaders) to attack
step forward in gluten-free world of baking. these peptides. Once activated, the T-cells also
recognize the patient’s own intestinal tissue, leading
COCKTAIL FODDER: GLUTEN- to significant damage and discomfort.
FREE MARKET • Gluten sensitive patients, on the other hand,
Market research company Packaged Facts predicts mount an innate immune response towards gluten.
that the gluten-free food market will grow to $6.6 billion This means while they do not produce gluten-
by 2017. specific antibodies or activate gluten-specific
T-cells, their immune system recognizes gluten
EASILY CONFUSED: CELIAC DISEASE peptides as foreign and potentially harmful, and
VS. GLUTEN SENSITIVITY activates generalized inflammatory pathways
Celiac disease and gluten sensitivity both involve an aimed at getting rid of them. This inflammation
immune response against peptides derived from gluten can cause symptoms similar to those seen in
proteins, but the details of that response differs. celiac disease, but there is no direct damage to the
intestinal tissue.
• Celiac disease patients mount what is called an
antigen-specific immune response. This means The two disorders can be differentiated by testing for
they generate antibodies that specifically recognize celiac-specific antibodies.
gluten peptides, and activate T-cells (a type of white

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BLUNT APPROACH: FECAL TRANSPLANTS Life Science Training from Industry Experts

FECAL TRANSPLANTS: A in developing a Crohn’s disease treatment based on an

BLUNT APPROACH anti-inflammatory protein originating from Bacteroides
thetaiotaomicron. Although this approach may be
The premise behind microbiome-based therapeutics is
more in line with familiar development and approval
straightforward: identify differences in the microbiome
pathways, in some cases these products may not be
of people suffering from a particular disease, and then
sufficient in treating microbiome-associated diseases,
try to make the sick person’s microbiome more like the
since drugs produced in this way will lack the ability
healthy person’s.
to colonize the intestinal tract as an actual bacterium
Rankings from the most radical (or just plain stomach- would. Further, there may be important and undefined
turning) to the most standard, here is a snapshot of interactions happening between different communities
microbiome-based therapeutics: of bacteria that a single protein will not be able
Fecal transplants represent the blunt approach of to replicate.
entirely resetting a patient’s microbiome with a
presumably healthy one. There are a few companies HEALTHY, BUT NOT THE CURE: YOGURT
trying to standardize this approach, including CIPAC We have heard the commercial, “Activia helps regulate
(Laguna Beach, CA), and Rebiotix (Roseville, MN). Both the digestive system when consumed daily for two
companies are working to develop standardized, pre- weeks as part of a balanced diet and healthy lifestyle.”
screened fecal matter as a therapeutic for C. difficiles Activia and food products marketed as “probiotics”
infection—unfriendly bacteria that cause severe are sold at a premium because they claim to promote
diarrhea, affecting several million people annually and the growth of friendly bacteria. While these products
leading to thousands of deaths. certainly cannot hurt and may be an important
Most people harbor low levels of C. difficiles without part of a healthy diet, they are simply not potent
a problem. The use of antibiotics, however, can enough to benefit individuals who are sick from
wipe out the friendly bacteria that keep C. difficiles microbiome imbalance.
in check. The current standard of care is treatment
with vancomycin—an antibiotic that many strains
MOSTLY HUMAN?
of C. difficiles are becoming resistant to. Resetting Every surface and crevice of your body is teeming with
the microbiome may be a safer and more effective microbes. Think about that buffet you had for lunch—
approach for the long term. how many of those microbes from the person in front of
you fell into the very same dish you took your scoop of
Other companies are working on a more targeted
food from?
approach: defining and introducing groups of specific
bacterial strains (a consortia) likely to be beneficial in a Your respiratory and gastrointestinal tracts have
particular disease state. This approach enables greater also been been colonized, playing host to thousands
specificity and quality control. Vedanta Biosciences of different species. Did that buffet line include
(Boston, MA) exemplifies this approach in their any sneezers?
identification of bacterial strains that have been shown In fact, bacterial cells in our body outnumber human
to suppress inflammation and are decreased or missing cells by about ten to one. Bacteria alone contribute
in patients with chronic gut inflammation. a few pounds of weights for every 100 pounds of
An even more targeted approach is to isolate a human. Yet most of us go about our daily lives with
protein or small molecule being secreted by a strain of no awareness of these invisible passengers, much
bacteria that gives it its therapeutic effect, and develop less an appreciation of their vital interactions with our
only that molecule as the therapeutic. GT Biologics own cells.
(Aberdeen, United Kingdom) has taken this approach

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Until recently, scientists (like those of us in the COCKTAIL FODDER:
buffet line) have also been guilty of ignoring this MICROBIOME DIAGNOSTICS
bacterial community.
The differences in microbiomes between healthy and
Medicine has largely been concerned with the very sick people are being used as diagnostic biomarkers.
small percentage of bacteria that cause diseases such As data on the specific genomic signatures of the
as pneumonia. That paradigm is changing quickly. microbiome become better characterized, companies
On an almost weekly basis, new findings inform us of such as Enterome (Paris, France) and Metabogen
these microorganisms’ role in regulating the healthy (Gothenberg, Sweden) are developing diagnostics based
functioning of important processes such as digestion on detecting these differences.
and immunity, and their role in keeping disease-causing
bacteria in check. TERM OF THE WEEK: METAGENOMICS
Imbalances within the microbiome have been linked to The human microbiome is the aggregate of
a range of disorders including obesity, inflammatory microorganisms (bacteria and fungi) that reside on
bowel disease, type 2 diabetes, anxiety, and depression. the surface and in deep layers of skin, in the saliva
What does this mean for biopharma? In this issue, we and oral mucosa, on the surface of the eye, and in the
explore some early stage approaches to translating our gastrointestinal tracts.
growing knowledge of the microbiome into mainstream Characterizing the microbiome is done in part through
clinical treatments and diagnostics. metagenomics, or the study of genetic material
recovered directly from samples as opposed to grown
in the lab.

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SCIENCE GEEKS ROCK! Life Science Training from Industry Experts

THE SCIENCE DRIVING BIOTECH this talk together was limiting it to just ten topics! Two of
the topics that I chose are summarized below:
Next week, BioTech Primer will be participating the BIO
2014 Annual International Convention in San Diego, Genome Editing: we often refer to a person’s genome—
CA. The event draws thousands and, as in years past, his complete set of genes—as a blueprint for life.
BioTech Primer will be headlining a few events. Join us Sometimes, there are errors in this blueprint. Wouldn’t
and learn! it be great if we could edit the blueprint and correct
these errors? We might soon be able to. Researchers
On Monday, June 23, I will be teaching an all-day class
have developed tools that make it possible to cut an
called “The Science Driving Biotech.” What can you
individual’s DNA in a specific location. Cellular enzymes
expect to learn?
then attempt to repair this break, but typically make
Starting with the basics: what is DNA? How do cells use mistakes in this repair, rendering the gene ineffective.
DNA to make proteins? How has the biotech industry This approach can be used to “knockout” a defective
adapted this knowledge to make biologic drugs? I will gene. This may sound like science fiction, but Sangamo
then quickly move into more complex, industry-relevant BioSciences is currently conducting Phase II human
applications and products. clinical trials testing this technology’s ability to render
Some of my favorite topics to teach include the genetic human T-cells resistant to HIV. Sangamo also has
variation, genomics, and stratified medicine sections positive preclinical data supporting the idea of using
of the course. Building on the concept of the gene as this technology to actually correct a gene sequence by
the basic unit of human inheritance, I love to surprise co-delivering a “repair template”—a short piece of DNA
my students with information on just how highly containing the correct gene sequence which the cellular
similar we are at the level of DNA sequence—but repair enzymes can then use to correct the defective
how in some cases, very small differences can lead gene sequence.
to a disease state. We then move on to a discussion Immunotherapy: the term “immunotherapy” covers a
of stratified medicine—how disease-related genetic wide range of topics, including vaccinations, monoclonal
information is being used to develop therapeutics that antibody therapeutics, stimulating a patient’s own
are targeted at specific subsets of patients and the immune system to fight cancer or other diseases,
companion diagnostics being developed to identify and using antibodies to deliver toxic compounds
the patients most likely to respond favorably. Finally, directly to cancer cells. During Tuesday’s talk, we
I give an overview of the next-generation sequencing will give an overview of what vaccines are, how they
technologies that are being used to generate this work, and describe promising work that may lead to
genetic data. an HIV vaccine. We will also discuss a personalized
This is a BIO2014 affiliate event and registration is open cancer vaccine developed using a patients’ own cancer
to everyone. The class will take place at Manchester cells, and describe a therapeutic vaccine designed to
Grand Hyatt San Diego. If interested click here for more activate the body’s own immune response to cancer.
information and registration. Due to limited space, Finally, we will talk about some of the new and exciting
preregistration and payment are necessary. I hope to applications of monoclonal antibodies from targeting
see you there! Alzheimer’s disease to use as a vehicle for drug delivery
in antibody-drug conjugates.
2 OF THE TOP 10 Please join me and Collins on Tuesday morning, June
Next week, I will be joined by my fellow BioTech Primer 24th, from 9:00-11:00 at the San Diego Convention
instructor Dr. Collins Jones to deliver a two-hour talk Center in room 23ABC. This talk is only open to BIO2014
on the Top 10 Current Science Trends & Advances at registrants. FYI: Last year the room was filled to
BIO2014 in San Diego. The hardest part about putting

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capacity and many were turned away at the door. Get target, many validation assays attempt to measure
there early and get your seat! the effects of inhibition. In some cases, a selected
target may play a role in disease progression – but if
NO SCIENCE SINCE HS? WE CAN HELP it is inhibited, another cellular protein will simply take
BioTech Primer has published a book titled The Biotech its place, nullifying the potential therapeutic effect
Primer: An insider’s guide to the biotech and pharma of an inhibitor. In other cases, inhibiting a selected
industry—though some refer to it as “Biotech for target may have the desired therapeutic effect—
Dummies.” The 200-page book explains the science halting cancer cell growth, for example—but may also
behind the biotech industry, and includes a glossary result in unexpected side effects, such as the death of
of commonly used terms. Similar to our classes, healthy cells.
The Biotech Primer starts out with the fundamentals One of the most popular ways of testing the effects
of biology used by researchers and progresses to of inhibition in cell-based assays is through the use of
how those fundamentals are employed to create RNAi, described in detail in the chapter “From Gene to
therapeutics. The text is written to be understood by Protein.” RNAi is an effective way to quickly determine
all—even those who have not taken a science class since the results of blocking the production of a particular
high school. The illustrations and cocktail fodders (so protein, thus mimicking the effects of a strong inhibitor.
you can impress your friends at your next party) keep
If the cell models show promise, the researchers will
things interesting. The Biotech Primer will be available
move on to animal models, most likely designing
all next week at the BIO Bookstore. If you are not
experiments using so-called “knockout” mice—
going to San Diego, but would like a copy, click here to
mice in which a particular gene has been disrupted.
order online.
Researchers can ask similar questions to those asked
Excerpted below are a few paragraphs from Chapter 8: in the cell model, but on the scale of the whole animal:
The Science of Discovery do the experimental mice still get cancer, Parkinson’s
Validating The Target disease, diabetes, or heart disease when the target
gene is silenced or absent? The animal model also
Once a potential drug target has been identified,
provides valuable information about targeting safety
researchers will try to validate the target by determining
that might not be addressed in cell models because it is
whether the target plays a key role in the disease
possible to examine the effects of gene targeting on the
process and whether targeting it is likely to be both
whole organism.
safe and effective. Target validation is a very important
step in the drug discovery process, since research and
COCKTAIL FODDER
development gets progressively more expensive—if a
drug is unlikely to be successful, millions of dollars can The introduction of genetic information from spiders
be saved if this is realized early on. into goats allows these mammals to produce spider
silk in their milk, which is collected and purified to
Target validation will most often include cell-based
make products such as soft body armor (89 The
assays (in vitro testing) and animal models (in
Biotech Primer).
vivo testing). Since the goal of many therapeutic
interventions is to inhibit the activity of the selected

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SCABBING NOW A “WALK IN THE PARK”
FOR HEMOPHILIA A SUFFERERS Life Science Training from Industry Experts

SCABBING NOW A WALK IN THE PARK OTHER FC-FUSION PROTEINS

FOR HEMOPHILIA A SUFFERERS ON THE MARKET
Imagine tripping over your feet, face-planting on a While Biogen is the first company to use this approach
sidewalk. You are in shorts, your knees scrape, and you for hemophilia therapeutics, there are several other Fc
are bloody. You clean it off, and soon you are the proud fusion proteins currently on the market.
owner of some new scabs. Life goes on. For example, rheumatoid arthritis (RA) drug Enbrel
Let’s say you are the one of the 5,000 men with (Amgen/Pfizer) is a fusion of the tumor necrosis factor
hemophilia A—a condition preventing scab formation receptor (TNFR) and the Fc region of an antibody. The
caused by a lack of blood clotting factors. You would receptor soaks up immune system-activating tumor
need to start replacement therapy right away, necrosis factor before it can fully activate the immune
obtaining the missing protein factors by donor plasma response, relieving the symptoms of RA. Enbrel has
or recombinant DNA technology. If you do not, the also been approved for psoriasis. Eylea (Regeneron)
bleeding could last days, weeks, or even cause fatality. fuses an antibody Fc region with VEGFR, the receptor
Although effective, traditional replacement therapies for vascular endothelial growth factor; Eylea soaks up
have been cumbersome in practice, especially for VEGF, inhibiting blood vessel growth in the treatment of
patients with severe hemophilia. This is due to the wet macular degeneration.
clotting factors’ instability. After infusion, the factors As with the hemophilia treatments, fusion with the
have a half-life of between 10 and 25 hours. Fc region makes these proteins more stable in the
Thanks to the FDA’s approval of Biogen Idec’s Eloctate patient’s blood stream.
that occurred last week, hemophilia A patients have a
considerably easier time acquiring scabs. Eloctate is NATURALLY OCCURRING
also a type of replacement therapy, but with a twist. FUSION PROTEIN
It is an “Fc Fusion Protein”—a hybrid protein that Most fusion proteins are created in the lab for
consists of the clotting factor fused with the Fc region therapeutic purposes. However, one very well
of an antibody. The Fc region is a structural feature of characterized fusion protein, Bcr-Abl, occurs in patients
antibodies that gives them their unusually high stability, with chronic myelogenous leukemia (CML). Bcr-Abl is
enabling them to remain in circulation for as long as the result of a chromosomal translocation, an event in
several weeks. Fusing the Fc region with clotting factors which parts of two chromosomes swap places, fusing
buys patients a few days of stability—long enough together the genes “Bcr” and “Abl.” The result of this
to reduce the frequency of administration to once fusion is the chronic activation of an enzyme that turns
every four days. Given that most receive factor VIII on cell division. Gleevec (Novartis) inhibits this mutant
treatment as an intravenous infusion, this advancement protein and is approved for the treatment of CML.
promises significant quality of life improvements
for those patients requiring prophylactic treatment COCKTAIL FODDER
for hemophilia.
Hemophilia is sometimes referred to as “the royal
Back in March of 2014, Biogen won approval for Alprolix, disease” because of its prevalence in the European
another fusion protein product (Fc-Factor IX) for the royalty of the 19th and early 20th centuries. Those
treatment of hemophilia B. afflicted were all descendants of Queen Victoria—
and royal families of Spain, Germany, and Russia
were afflicted.
In 2009, scientists at the University of Massachusetts
analyzed DNA from bone fragments of the Romanovs

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(the last Russian royal family) and found a mutation Fusion proteins are created by combining two different
suggesting “the royal disease” was indeed hemophilia B. genes (or recipes) to produce one final product.
Scientists use the enzyme DNA ligase to make the
TERM OF THE WEEK: DNA LIGASE magic happen. DNA ligase can be thought of as
Proteins are created from recipes — what we all know molecular glue.
as genes.

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SAVED BY SCREENING Life Science Training from Industry Experts

NEW COMPANION TERM OF THE WEEK:

DIAGNOSTIC APPROVAL COMPANION DIAGNOSTIC
Colorectal cancer is the third most common cancer A companion diagnostic is the test or measurement
found in both men and women in the U.S., and is the intended to assist physicians in making treatment
second leading cause of cancer deaths. Approximately decisions for their patients, usually by determining the
1.2 million cases of colorectal cancer are expected to efficacy and/or safety of a specific drug for a targeted
occur globally. patient group.
Just last week, the FDA approved Qiagen’s Therascreen Here’s a list of all FDA approved companion diagnostics so far.

KRAS companion diagnostic to identify colorectal

cancer patients most likely to respond to Amgen’s HOW IT WORKS:
Vectibix. This approval follows a 2012 approval of THERASCREEN DIAGNOSTIC
the same test as a companion diagnostic for Eli
The Therascreen test works by taking DNA from a
Lilly’s Erbitux.
patient’s tumor cells and mixing it with reagents that
Vectibux and Erbitux both target epidermal growth specifically bind to and make copies of the tumor’s
factor receptors (EGFR), which is overexpressed in some KRAS gene.
types of colon cancer.
If a specific cancer-associated mutation is present, a
EGFR is found on the cell surface and is activated by fluorescent signal is released. Therascreen tests for the
growth factor binding. Once activated, EGFR activates presence of seven specific mutations known to make
enzymes inside the cell that drive the cell forward EGFR inhibitors ineffective.
into cell division. Vectibix and Erbitux both work by
inhibiting EGFR’s activation. COCKTAIL FODDER
About 50% of colorectal patients respond well to According to research and marketing firm GlobalData,
treatment with an EGFR inhibitor. Those that do not the companion diagnostics testing market value will
respond usually have a mutation in a “downstream” increase from $421 million in 2013 to $670 million
enzyme—one of the enzymes driving cell division that is by 2020.
normally only activated in response to EGFR activation.
One such enzyme that is commonly mutated in colon LEARN MORE: EGFR AND CANCER
cancer is called KRAS. Therascreen detects KRAS EGFR overexpression is associated with a number
mutations that make a patient non-responsive to EGFR of cancers, including lung cancer, anal cancers, and
inhibitors such as Vectibix and Erbitux. glioblastoma multiforme.
Given the high cost of these drugs—up to $30,000 These somatic mutations involving EGFR lead to its
per eight weeks of treatment—this new companion constant activation, which produces uncontrolled cell
diagnostic should be welcome by insurers and patients division. Mutations, amplifications or misregulations of
alike, helping to accurately treat colorectal cancer EGFR or family members are implicated in about 30%
patients by eliminating a costly and time consuming of all epithelial (tissues that line cavities and surfaces of
drug therapy for patients who would not see any structures throughout the body) cancers.
benefit from it.

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MRSA VS. DALVANCE: LET’S
GET READY TO RUMBLE Life Science Training from Industry Experts

MRSA’S MATCH FDA QIDP STATUS GRANTED

Last week, the FDA approved Dalvance, Chicago-based Dalvance is the first drug to be approved under the
Durata Therapeutic’s new antibacterial drug for the FDA’s new Generating Antibiotic Incentives Now (GAIN)
treatment of methicillin-resistant staphylococcus program, established as part of the FDA Safety and
aureus (MRSA) infections. Innovations Act of 2012. Under GAIN, antimicrobials
Why is Dalvance newsworthy? MRSA is an in development may be granted Qualified Infectious
infectionthatis difficult to treat because it is resistant Disease Product (QIDP) status. QIDP status enabled
to all the usual medications: the cephalosporins and Dalvance to get priority review, as well as an additional
penicillins (methicillin, dicloxacillin, oxacillin, etc.) five years of data exclusivity now that it is approved.

What is MRSA? MRSA skin infections may begin as In the past decade, drug-resistant and multi-drug
uncomfortable sores or boils on the skin’s surface resistant bacteria strains have become one of the
that if left untreated may progress to deep, painful greatest public health challenges of the twenty-
abscesses requiring surgical draining. MRSA infections first century. Antibiotic resistance occurs when a
that enter the bloodstream or joint tissue can be life- few bacteria in a given population develop a genetic
threatening. mutation that makes them able to survive, even in
the presence of a particular antibiotic. For example,
Who is most susceptible to MRSA? MRSA is a serious
suppose an antibiotic works by inhibiting an enzyme
threat to the hospitalized, potentially infecting surgical
required for bacterial replication. If one bacterium
wounds or gaining entry into a patient’s bloodstream
mutates so the enzyme now has a slightly different
through an invasive device such as a catheter. MRSA
shape and is therefore no longer inhibited by the
outbreaks are also reported in prisons, shelters,
antibiotic, it will continue to reproduce while all
military barracks, and nursing homes because of
the other bacteria die off. Over time, this resistant
crowded and confined living conditions. Approximately
type becomes the dominant strain, spreading from
2% of the population carries MRSA on their skin and
person to person, unchecked by antibiotics. This type
although most do not exhibit signs of infection, they can
of resistance is caused in large part by overuse of
pass it on to individuals with open wounds or weakened
antibiotics—the more a bacterial population is exposed
immune systems.
to antibiotics, the greater the chance that resistance
How does Dalvance work? It works by inhibiting will develop.
bacterial cell wall synthesis. The cell wall is a layer of
Each year in the U.S., at least 2 million people become
sugars and amino acids (peptidoglycans) that surround
infected with bacteria that are resistant to antibiotics
bacterial cell membranes, providing bacteria with
and at least 23,000 die as a direct result of these
structural support and protection, as well as acting
infections (source: CDC). Many more die from other
as a filtering mechanism. Dalvance is a synthetic
conditions that are complicated by an antibiotic-
“lipoglycopeptide”—a chemical entity similar enough to
resistant infection. In addition to MRSA, other drug-
the peptidoglycans incorporated into the cell wall, but
resistant microorganisms of urgent concern include
different enough so that once incorporated, cell wall
Clostridium difficile (life-threatening diarrhea),
synthesis stops. Without a functional cell wall, bacteria
Enterobacteriaceae (bloodstream infections), Neisseria
die. Human cells do not have cell walls, therefore they
gonorrheoeae (severe reproductive complications),
are not affected.
Pseudomonas aeruginosa (pneumonia, bloodstream,
urinary tract, and surgical site infections), and
Mycobacterium tuberculosis (tuberculosis). Hopefully,
the GAIN program will lead to the discovery and

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development of a wide range of novel antimicrobials to COCKTAIL FODDER
fight these emerging threats.
Fungi have been an important source of antimicrobials.
BACTERIA VS. VIRUSES Penicillin was discovered in 1928 when microbiologist
Alexander Fleming observed that infection of his
Most of us have been told by our family doctor that
bacterial cultures with the Penicillium fungi inhibited
a prescription for antibiotics will do no good against
bacterial growth. Penicillin was hailed as a wonder
a viral infection such as the flu or the common cold.
drug, but as early as the 1950s—just a few years after
During flu season, instead of taking a few rounds of pills
widespread use began—problems with bacterial
and starting to feel better, we have to suffer through
resistance emerged. Despite the many lives penicillin
a week or so of sore throat and exhaustion, aided only
has saved, it is also the most common drug allergy,
by chicken soup and extra rest before getting back to
illustrating why it is important to have a wide range of
normal. Why is this?
potential antibiotics on hand.
Bacteria are independent, single-celled organisms with
their own cell membrane, cell wall, and DNA/protein- IN THE PIPELINE
making machinery. Bacteria do not need another living Massachusetts-based Cubist Pharmaceutical’s new
organism to survive. Most antibiotics work by targeting antibiotic, Sivextro—also being developed for MRSA
structures such as cell walls that are unique to bacteria skin infections—has received the QIDP designation.
so the bacteria cells are killed, but human cells are Sivextra was unanimously recommended by an FDA
not harmed. advisory panel last March; a final approval decision
In contrast, viruses are not independently living is expected by June 20th. Sivextra works by inhibiting
organisms. Most consist of only some genetic material the bacterial ribosome - the enzyme that makes all
(DNA or RNA) encapsulated in a few proteins and bacterial proteins. Without new protein production,
sometimes a membrane. They are not able to fully the bacterium is unable to carry out functions essential
replicate their genetic material or make the proteins for life, and dies. Cubist Pharmaceuticals has another
they need for survival without the aid of another living product in Phase III development for C. difficile-
cell–its host. This means it is more difficult to make a associated diarrhea.
drug targeting just the virus without also harming its
host—us.

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DON’T BE DOA. LEARN THE MOA. Life Science Training from Industry Experts

MAJOR MECHANISMS OF ACTION (November 2007), and to treat irritability in children

with autism (November 2009).
The top selling drugs of 2013 were Abilify (Otsuka
Pharmaceuticals; $6.9 billion), Nexium (AstraZeneca; #2: NEXIUM
$6.3 billion), and Humira (AbbVie; $5.9 billion). Anyone
AstraZeneca’s Nexium is a small molecule
who follows biopharmaceuticals—and plenty of people
drug prescribed primarily for the treatment of
who do not—are familiar with these drugs.
gastroesophageal reflux disease (acid reflux) and ulcers.
How do they actually work?
Nexium reduces the acidity of the stomach by inhibiting
In this issue, we will take a look at the diverse an enzyme called a proton pump which is present on
mechanisms of action (MOA) of these top-selling drugs. the surface of the cells that line the stomach.

#1: ABILIFY Proton pumps do exactly what their name implies—they

pump protons, or positively charged hydrogen atoms
Otsuka’s Abilify is a small molecule drug first approved
(H+) into the stomach where they combine with chloride
in 2002 used to treat schizophrenia, a disease which
ions (Cl-) to make hydrochloric acid (HCl).
is not fully understood but involves irregularities in
dopamine signaling. HCl plays an important role in the digestion of food,
helping to break down proteins and other nutrients
Schizophrenics typically have elevated levels of
so they can be absorbed in the intestine. However,
dopamine signaling in the mesolimbic system—the part
in individuals with defects in the barrier between
of the brain associated with reward and desire. These
the esophagus and the stomach, acid moves into the
elevated levels are thought to be responsible for the
esophagus, causing heartburn and food regurgitation,
positive symptoms of schizophrenia: visual or auditory
sometimes accompanied by nausea, pain with
hallucinations, distress, and agitation. At the same time,
swallowing, and chest pain. Decreasing stomach acid
they exhibit lower dopamine levels in the mesocortical
production relieves these symptoms. Likewise people
pathways—the part of the brain involved in emotional
with ulcers—open sores in the stomach or intestine
response, motivation, and cognitive control. These
usually caused by infection with the bacteria H. pylori—
lower mesocortical levels are thought to be responsible
find relief from pain with decreased stomach acid.
for the negative symtoms of schizophrenia: lack of
motivation and emotional response. #3: HUMIRA
Abilify works by modulating this complex dopamine AbbVie’s Humira, the third top-selling drug of 2013,
system. It is a partial agonist of the dopamine D2 is a monoclonal antibody designed to target the pro-
receptor, meaning it activates the receptor, but not to inflammatory signaling molecule tumor necrosis factor
the same extent that dopamine does. In the regions of alpha (TNF-alpha).
the brain where there is too much dopamine activity,
TNF-alpha’s normal function is to turn on the body’s
Abilify competes with dopamine for receptor binding,
inflammatory response which is the migration of
resulting in less activation. In regions of the brain where
several different types of white blood cells towards a
there is too little activity, Abilify provides at least some
site of infection or injury within the body. Symptoms
activation. Earlier anti-psychotics simply blocked the
such as swelling, heat, and pain occur, but it ultimately
activation of dopamine receptors, and thereby did little
rids the body of the infectious agent or repairs
to ameliorate the negative symptoms of schizophrenia.
damaged tissue.
Abilify has also been approved for acute manic episodes
In a healthy individual, this inflammatory response goes
associated with bipolar disorder (October 2004), as
away after the infection or injury is resolved. In people
an adjunct therapy for major depressive disorder
with autoimmune disorders, this response is activated

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by the patient’s own tissues – thus the inflammation blood cells, thereby “turning down” the inflammatory
doesn’t resolve on its own, leading to chronic pain and response. Humira has been approved for a wide
loss of function. range of autoimmune disorders including rheumatoid
Humira works by binding TNF-alpha and preventing arthritis, psoriatic arthritis, ankylosing spondylitis,
it from activating its receptor on the surface of white Crohn’s disease, and ulcerative colitis.

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MITOCHONDRIAL MEDICINE
GOING MAINSTREAM? Life Science Training from Industry Experts

MORE THAN “JUST” A POWERHOUSE discover and develop drugs targeting mitochondrial
function. Likewise, Edison Pharmaceuticals (Mountain
When we hear the word mitochondria, most of us
View, CA) entered an alliance with Dainippon Sumitomo
remember it described in high school biology class as the
Pharma, also known as DSP (Osaka, Japan) to develop
“powerhouse” of a cell. It is an apt moniker, and not just
drugs for inherited mitochondrial diseases. Could
because mitochondria are the subcellular compartments
these companies push mitochondrial medicine into
that convert glucose from our diet into chemical energy
the mainstream?
that our cells can use to do work. These powerful
compartments also play a role in such diverse cellular ALZHEIMER’S & MITOCHONDRIAL
processes as cellular differentiation, cell death, and FUNCTION: CAUSE OR EFFECT?
cell signaling.
Alzheimer’s disease (AD) is linked to mitochondrial
It is not surprising that defects in mitochondrial function dysfunction; however, scientists have been unsure as
are associated with a whole range of diseases, including to whether or not this was a cause or a symptom of
musculoskeletal diseases, metabolic diseases, and the disease.
neurodegenerative diseases.
Recent studies suggest that mitochondrial dysfunction
NEW DRUG TARGETS? may precede Alzheimer’s rather than the other way
around. Swedish and German researchers report
Mitochondrial biology is interesting to drug researchers
increased aging (including neuronal degeneration)
because they contain their own DNA, distinct from the
in mice engineered to have increased levels of
DNA found in the nucleus of human cells. Until recently,
mitochondrial DNA mutations. A group of Spanish
scientists assumed mitochondrial DNA mostly codes for
researchers reported finding decreased levels of
enzymes involved in carrying out the chemical reactions
mitochondrial DNA in the cerebrospinal fluid of
needed to provide cellular energy. It turns out that
people developing AD. This is a good predictor of
mitochondrial DNA may also hold the recipe for proteins
the disease onset, suggesting causative role for
involved in various diseases.
mitochondrial irregularities.
For example, we now know certain versions of
These findings suggest a potential new way of thinking
mitochondrial genes are associated with an increased
about Alzheimer’s treatment. Current studies mostly
or decreased risk of stroke. Preliminary studies also
focus on reducing the primary physiologic hallmark of
suggest a possible link between certain mitochondrial
the disease - amyloid-beta (Aß) protein plaque formation
genes and other complex diseases such as Parkinson’s,
in the brain. However, clinical trials of monoclonal
Alzheimer’s, and diabetes, opening up exciting new
antibodies targeting the amyloid-beta (Aß) protein
possibilities for understanding human disease. It is also
plaques and of inhibitors of the enzyme beta-secretase
an untapped target for drug discovery.
(whose activity is required for the formation of Aß
plaque) have been disappointing. The ability to detect
COMPANIES TO WATCH and treat abnormalities in mitochondrial function before
Mitochondrial medicine is still largely at the basic the onset of symptoms may prove to be a game-changer
research stage, as investigators strive to better for this challenging disease.
understand what role these tiny powerhouses play in
a range of diseases. However, a few companies—both MITOCHONDRIAL DNA
start-ups and established players alike—are beginning to REPLACEMENT: IVF IMPLICATIONS
take note of the field’s potential for new drug discovery.
Mitochondrial DNA is inherited maternally and this has a
Last fall, Mitokyne (Cambridge, MA) announced a new range of implications for IVF.
partnership with Astellas Pharma (Tokyo, Japan) to

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Why isn’t mitochondrial DNA also inherited paternally? egg produced by one woman is implanted into another
Only the ova or egg (not the sperm) contain a supply woman who is unable to produce healthy eggs and
of mitochondria. therefore cannot get pregnant.
Why does only the egg contain mitochondria? If the egg Donor egg is a proven technology—the first successful
is fertilized by the sperm, the mitochondria grow and birth via donor egg occurred in 1983, making it nearly
replicate along with the resulting zygote (fertilized egg), as old as the biotechnology industry itself. With a better
thus the mitochondria are passed on in subsequent understanding of genetics, patients are demanding
rounds of cell division. refinements to this popular procedure. One includes
What implications does this have for the infertility transferring the healthy nuclear DNA from the egg of
industry? It is estimated that over five million babies a patient with defective mitochondrial DNA into the
have been born since the very first in vitro fertilization enucleated egg of a donor, and then implanting that
(IVF) procedure which occurred 36 years ago. IVF is a “hybrid” donor egg back into the patient. Currently,
general term which spans many different procedures the FDA is considering what guidelines might be
including a donor egg procedure. A donor egg is the appropriate for this new technique.
process of third party reproduction in which a healthy

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GENOME EDITING: CURING HIV? Life Science Training from Industry Experts

CURING HIV? when replicating itself. If the virus cannot get into the
modified T-cells to replicate, then these mutations will
HIV destroys its victim’s immune system by infecting not arise.
T-cells, a type of white blood cell critical for immunity.
HIV binds to two different proteins on the T-cell’s GENOME EDITING: HOW IT WORKS
surface: CD4 and CCR5.
Scientists are currently researching multiple ways of
In the late 1990s, scientists identified a population editing genomes, but the approach furthest along
of individuals seemingly resistant to HIV infection. in clinical development is called ZFN, or “zinc finger
Despite multiple known exposures, they did not nuclease” technology. ZFNs are proteins that have been
get infected. It turned out that these HIV-resistant engineered to recognize a unique DNA sequence and
individuals had something in common: a mutation in cut the DNA in that location. ZFNs are able to recognize
the gene that coded for the CCR5 protein, resulting in longer DNA sequences than other approaches, such
a shortened version of the protein appearing on their as restriction enzymes, enabling scientists to engineer
T-cell’s surface. As a result, HIV simply was not able to ZFNs that will cut at a unique location within the
efficiently infect their T-cells, so they did not get sick. human genome.
The mutated CCR5 protein appears to have no negative
Once the DNA is cut, cellular enzymes attempt to
affect on T-cell function.
repair the cut, disrupting the gene in the process. This
Today, Sangamo Therapeutics is applying ZFN genome- approach is currently being tested in Phase 2 clinical
editing techniques to disrupt the CCR5 gene of HIV trials as a potential HIV treatment (see “Curing HIV?”).
patients’ T-cells. This stops the virus from infecting new Researchers are also developing methods to deliver
cells and restores the immune system. The therapy is a DNA “repair template” along with the ZFN. In this
currently in Phase 2 clinical trials. These are the first scenario, the cellular repair enzymes will incorporate
clinical trials of any sort of genome editing. the correct version of the gene at the cut site; rather
than simply disrupting a dysfunctional gene, they will
THIS TIME, IT IS DIFFERENT repair it. ZFN-mediated repair of the hemoglobin gene
The idea to knock out the CCR5 protein in the T-cells in sickle cell anemia is currently in preclinical trials.
of HIV-infected patients came from the 2009 reports
of the “Berlin Patient”—an HIV patient who received a COCKTAIL FODDER
bone marrow transplant for the treatment of leukemia What do smallpox and HIV have in common? Four
from an individual with the protective CCR5 mutation. percent (4%) to 16% of people of European descent
After receiving the transplant, the patient went off his have the CCR5 mutation, protecting them against HIV
antiviral medications and remained virus free. infection. Interestingly, the smallpox virus—which
The genome editing treatments hope to mimic this ravaged Europe from the middle ages until a vaccine
success. By isolating T-cells from HIV-infected patients, was discovered in 18th century—also uses CCR5
disabling the gene for the CCR5 protein, and infusing to infect cells. It is thought that the CCR5 mutation
these edited cells back into the patients, doctors will persisted due to its protection against smallpox.
create a population of HIV-resistant cells within the
TERM OF THE WEEK:
patient. The virus will continue to infect and kill off the
RESTRICTION ENZYME
unmodified cells, leading to a T-cell population that
consists only of HIV-resistant cells. Without a host, the A restriction enzyme is an enzyme that cuts DNA
virus will not be able to survive. at a specific nucleotide base (A,T,G, or C) sequence
known as restriction sites. Restriction enzymes occur
For decades, new anti-HIV treatments have been
naturally in bacteria where they serve as a defense
developed, only to have the virus evolve resistance to
against invading viruses. Hundreds of different types of
them. This is because the virus has an unusually high
restriction enzymes have been isolated from bacteria
mutation rate—changes to its genetic information
and are commercially available for use in genetic
happen very frequently due to mistakes that HIV makes
engineering applications.

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RNAI ADVANTAGE GOES TO DICERNA Life Science Training from Industry Experts

THE SCIENCE BEHIND THE thus blocking expression (or making) of a disease-
IPO CRAZE: OUR FOCUS FOR associated protein.
THE NEXT FEW WEEKS RNA is naturally single-stranded in the human body.
I want to thank you for subscribing to Biotech Primer When the body finds dsRNA, it recognizes something is
WEEKLY. Now, imagine yourself on the set of Jeopardy! wrong and destroys it. By introducing dsRNA into cells,
facing this clue: it is possible to destroy the messenger RNA (mRNA),
which carries the sequence that codes for the disease-
“We saw 45 in 2013, and there’s been 17 so far this year.”
associated protein. If mRNA is destroyed, the protein is
The correct response: “What is the number of not produced and the person is no longer ill. Tricky!
biotech IPOs?”
Alex quips: “That is correct! And there’s no sign of a LEARN MORE: ADVANTAGE RNAI
biotech industry IPO slowdown.” The most tantalizing potential upside of RNAi
Great news if you own one of these companies, and therapeutics is their ability to target previously
even better if you are a patient (and aren’t we all at “undruggable” targets.
some point). Investment equals innovation. Innovation Biologic drugs such as monoclonal antibodies can be
ultimately results in new therapeutics. The therapeutic highly effective against targets on the surface of a cell
areas bringing in the most IPO bucks are cancer, or circulating in the patient’s blood; however, they are
autoimmune disease, and rare diseases. physically too large to enter the patient’s cells. Small
In the next few issues, we will take a look at the science molecule drugs can enter a patient’s cells but it is
behind the buzz. not always possible to identify an appropriate small
molecule binding site on these intercellular targets.
IPO FOCUS: DICERNA Potential drug targets such as transcription factors—
the proteins that “turn on” the production of a specific
One of the hottest IPOs of 2014 is Dicerna
protein or group of proteins in response to a particular
Pharmaceuticals. Staying true to the IPO trend, Dicerna
signal—have been difficult to reach with either small or
focuses on both cancer and rare disease therapeutics.
large molecule therapeutics.
Offered at $15/share, the company sold six million
shares, raising $90 million in cash. The success of the Various types of cancer have been associated with
initial offering exceeded expectations and continues to overactive transcription factors which represent a
do well. In the ensuring weeks, shares traded as high as prime target for RNA-based therapeutics.
$44.95/share.
COCKTAIL FODDER
Dicerna claims to have a new and improved version of
RNA interference (RNAi), the gene-silencing technology The company name Dicerna is derived from the cellular
similar to the antisense technology featured in this enzyme that activates the RNAi pathway: Dicer.
space last month. The RNAi process works by taking
advantage of pre-existing cellular pathways that
TERM OF THE WEEK: DRUG TARGET
recognize and destroy double-stranded RNA (dsRNA), A drug target is a molecule involved in a disease that is
modified or affected by a potential therapeutic.

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NO FREE LUNCH FOR OBESITY DRUGS Life Science Training from Industry Experts

NO FREE LUNCH FOR OBESITY of BMS’s diabetes unit to AZ, Myalept has landed in
a new home just as it received FDA approval as an
A major lead in curing obesity came on the scene in the
orphan drug.
1990s when researchers noticed that obese mice lacked
a critical hormone dubbed leptin. TERM OF THE WEEK:
Suddenly, the tantalizing prospect of tackling obesity RECOMBINANT PROTEIN
with leptin treatments sent both drug developers and Recombinant protein refers to protein therapeutics, a
casual dieters astir. type of biologic. Drugs such as insulin, human growth
As the saying goes, there’s no free lunch, and hormone, or the newly-approved Myalept are examples.
excitement waned in recent years as it has become The gene that codes for the therapeutic protein—
apparent that unlike mice, most obese people for example, the LEP gene responsible for leptin—
experience a lack of response to leptin. is first isolated and then recombined with vector
It is not all bad news—people who have a rare genetic DNA in order to transfer it into the cell line to be
defect in leptin production now have the option of used for manufacturing. This technology is called
a drug. recombinant DNA technology and its final product is a
recombinant protein.
RECOMBINANT DRUG MYALEPT
APPROVED FOR LEPTIN DEFICIENCY COCKTAIL FODDER: LEPTIN
The FDA just approved Myalept for the treatment of The name “leptin” is derived from the Greek word
leptin deficiency on February 24, 2014. Myalept is a for thin.
recombinant version of leptin, and as such is considered The term “lipodystophy” is derived from the Greek
a replacement therapy—it is replacing a hormone words “lipo” for fat and “dystrophy” for abnormal or
(leptin) that the body should be making but is not. degenerative condition.
Leptin deficiency causes a rare disease known
as lipodystrophy, leading to widespread loss of ORPHANS OUTPACING OTHERS
subcutaneous fat resulting in multiple metabolic With the first quarter of 2014 nearly behind us, orphan
complications including severe insulin resistance. drugs have already won four of the five approvals from
Myalept, now part of AstraZeneca’s drug arsenal, was the FDA this year.
first developed by Amylin Pharmaceupticals, then Orphan drug approvals are outpacing all others in the
was picked up by Bristol-Meyers Squibb when they first quarter of 2014.
purchased Amylin in 2012. With the recent sell-off

Drug Name Innovator Approval Date Indication Orphan Drug?

Myalept Amylin Pharmaceuticals 2/24/2014 Lipidystrophy Yes

Northera Chelsea Therapeutics 2/18/2014 Neurogenic Orthostatic Hypotension Yes

Vimizim BioMarin Pharmaceuticals 2/14/2014 Morquio A Syndrome Yes

Hetlioz Vanda Pharmaceuticals 1/31/2014 Non-24 Hour Sleep-Wake Disorder Yes

Bristol-Meyers Squibb
Farxiga 1/8/2014 Type 2 Diabetes No
and Astra Zeneca

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200,000 IS THE MAGIC NUMBER Life Science Training from Industry Experts

ALPHABET SOUP: NEW The NDA application is the vehicle through which drug
DRUG APPROVALS sponsors formally propose that the FDA has approved
a new pharmaceutical for sale and marketing in the
Acronyms abound in the world of new drug approvals.
U.S. The data gathered during the animal studies and
Check out our cheat sheet below for help with some of
human clinical trials of an IND become part of the NDA.
the most common:

BLA: Biologic Licensing Application TERM OF THE WEEK: RARE DISEASE

Planning on developing a biologic drug? Would you like In the United States, the FDA considers disorders that
to bring it to market? You will need to submit one of affect fewer than 200,000 people a rare disease.
these to the FDA which gives them the details on your The term “rare disease” is often interchanged and
manufacturing process, chemistry, pharmacology, synonymous with orphan diseases. The latter term may
clinical pharmacology, and the medical effects of encompass both rare diseases and disorders that affect
your biologic. more than 200,000 people, but for which there is no
reasonable expectation for the cost of developing and
EMA: European Medicines Agency
marketing such a drug would be recovered.
Roughly parallel to the FDA, the EMA is the European
Union’s agency for the evaluation of new medicinal EASILY CONFUSED: BREAKTHROUGH
products. An EMA approval means market access to its VS. FAST TRACK DESIGNATION
27 member countries without having to seek approval Both designations are intended to expedite the
in each individually. development and review of drugs for serious or life-
IND: Investigational New Drug threatening conditions; however, there are differences
in what needs to be demonstrated for each designation.
Congratulations! There is a molecule you consider
a viable candidate for commercial development, To qualify for breakthrough therapy designation, the
understand its pharmacological properties, and have drug sponsor must provide preliminary clinical evidence
already run animal studies to determine it will not be indicating that the drug may demonstrate substantial
acutely toxic to human beings. Now it is time to contact improvement on a clinically significant endpoint(s) over
the FDA and fill out an IND, which upon approval, grants available therapies.
you the ability to begin limited human trials. In contrast, a drug may qualify for fast track designation
by submitting either nonclinical or clinical data
IRB: Institutional Review Board
that demonstrates the potential to address unmet
Once you are in clinical trials, an IRB—sometimes medical need.
known as an independent ethics committee or ethical
review board—is formally designated to approve, COCKTAIL FODDER: FIRST
monitor, and review biomedical and behavioral research BREAKTHROUGH DRUG
involving humans. The chief priority of IRBs is to protect Gazyva is a treatment for chronic lymphocytic leukemia.
human subjects from physical or psychological harm. It was approved by the FDA on November 1, 2013 and
NDA: New Drug Application has the distinction of the first drug with breakthrough
therapy designation to reach market.

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