DR MA OBIO

FMC path
THE CELL CYCLE
CONTENTS
LEARNING OBJECTIVES
INTRODUCTION
• CELL CYCLE
• Fundamentals of cell proliferation is the development, maintenance of steady-state tissue
homeostasis & replacement of dead or damaged cells.
• Key elements of cellular proliferation
• Accurate DNA replication
• Coordinated synthesis of all other cellular constituents
• Equal apportionment of DNA & other cellular constituents like organelles to daughter cells through
mitosis & cytokinesis.
• The sequence of events that results in cell division is called the Cell cycle (CC).
• CELL CYCLE
• The CC consists of:
1. G1 (Presynthetic growth) 2. S (DNA synthesis) 3. G2 (Premitotic growth)
4. M (Mitotic) phases 5. G0 state: Quiescent cells that are not actively cycling
• Cells can enter G1 either from the G0 quiescent cell pool or after
completing a round of mitosis.
• Each stage requires completion of the previous step as well as
activation of necessary factors.
• Non-fidelity of DNA replication or cofactor deficiency results in arrest
at the various transition points.
Cell-cycle landmarks
• Cell-cycle phases: G0, G1, G2, S & M
• G1 restriction point: Refers to the phase in G1 at which the cell gets
committed to the CC without further need of the growth factor that
initiated cell division.
• G1/S & G2/M cell-cycle checkpoints.
• Labile cells: Epidermis & GIT may cycle continuously
• Stable cells: Hepatocytes are quiescent but can enter the CC
• Permanent cells: Neurons & cardiac myocytes have lost the capacity to
proliferate.
REGULATION OF THE CELL CYCLE
• The CC is regulated by numerous activators & inhibitors.
• CC progression is driven by proteins called cyclins & cyclin-associated enzymes called cyclin-
dependent kinases (CDKs)
• CDKs acquire the ability to phosphorylate protein substrates (Kinase activity) by forming
complexes with the relevant cyclins.
• Transiently increased synthesis of a particular cyclin leads to increased kinase activity of the
appropriate CDK binding partner
• When the CDK completes its round of phosphorylation, the associated cyclin is degraded & the
CDK activity abates.
• It is like a relay race in which each leg is regulated by a distinct set of cyclins
• Over 15 cyclins have been identified
• Cyclins D, E, A & B appear sequentially during the CC & bind to one or more CDKs.
SURVEILLANCE MECHANISMS
• Are embedded in the CC, primed to sense DNA or chromosomal damage. These quality-
control checkpoints ensure that cells with genetic imperfections do not complete replication.
• G1-S check-point: Monitors the integrity of DNA before irreversibly committing cellular
resources to DNA replication
• G2-M check-point: Ensures accurate DNA replication before the cell actually divides.
• When cells detect DNA irregularities, checkpoint activation delays CC progression & triggers
DNA repair mechanisms.
• If the genetic derangement is too severe to be repaired:
• The cells either undergo apoptosis or enter a non-replicative state called senescence (Primarily
through p53-dependent mechanisms).
• Enforcing the CC checkpoints is the work of CDK inhibitors (CDKIs) by modulating CDK-cyclin
complex activity.
There are several different CDKIs
• One family of CDKIs composed of 3 proteins called p21 (CDKN1A), p27
(CDKN1B) & p57 (CDKN1C) broadly inhibits multiple CDKs
• Another family of CDKIs has selective effects on cyclin CDK4 & CDK6;
these proteins are called p15 (CDKN2B), p16 (CDKN2A), p18
(CDKN2C), p19 (CDKN2D)
• Defective CDKI checkpoint proteins allow cells with damaged DNA to
divide resulting in mutated daughter cells at risk for malignant
transformation
Biosynthesis of other cellular components
• This is another equally important aspect of cell growth & division
needed to make 2 daughter cells, like membranes & organelles.
• Thus when GF-receptor signaling stimulates CC progression, it also
activates events that promote changes in cellular metabolism that
support growth like the Warburg effect marked by:
• Increased cellular uptake of glucose & glutamine
• Increased glycolysis
• Decreased oxidative phosphorylation.
ROLE OF CYCLINS, CDKS & CDK INHIBITORS IN REGULATING THE CC.
• Shaded arrows represent phases of CC during which specific cyclin-CDK complexes are
active.
• Cyclin D-CDK4, cyclin D-CDK6 & cyclin E-CDK2 regulate the G1-to-S transition by
phosphorylating the Rb protein (pRb).
• Cyclin A-CDK2 and cyclin A-CDK1 are active in the S phase.
• Cyclin B-CDK1 is essential for the G2-to-M transition.
• Two families of CDK inhibitors can block activity of CDKs & progression through the CC.
1. “INK4 inhibitors,” composed of p16, p15, p18 & p19 act on cyclin D-CDK4 & cyclin D-
CDK6.
2. Three inhibitors (p21, p27 & p57) can inhibit all CDKs.
STEM CELLS
Totipotent stem cells: During development can give rise to all types of differentiated tissues
Adult stem cells: In the mature organism seen in various tissues only have the capacity to
replace damaged cells & maintain cell populations within the tissues where they reside.
• There are populations of stem cells between these extremes with varying capacities to
differentiate into multiple cell lineages.
• In normal tissues there is homeostatic equilibrium between replication, self-renewal &
differentiation of stem cells & the death of the mature fully differentiated cells
• The dynamic relationship between stem cells & terminally differentiated parenchyma is
seen by the continuously dividing epithelium of the skin.
• Thus, stem cells at the basal layer of the epithelium progressively differentiate as they
migrate to the upper layers of the epithelium before dying & being shed.
• Under conditions of homeostasis, stem cells are characterized by two
important properties
• Self-renewal: Permits stem cells to maintain their numbers may follow
asymmetric or symmetric division.
• Asymmetric division: Refers to cell replication in which one daughter cell
enters a differentiation pathway & gives rise to mature cells, while the
other remains undifferentiated retaining its self-renewal capacity.
• symmetric division: Both daughter cells retain self-renewal capacity.
Such divisions are seen early in embryogenesis (When stem cell
populations are expanding) & under conditions of stress, as in the bone
marrow following chemotherapy.
Mechanisms regulating cell populations
Cell numbers can be altered by:
1. Increased or decreased rates of stem cell input
2. Cell death from apoptosis
3. Changes in the rates of proliferation or differentiation
Embryonal stem cells (ES cells)
• The zygote (Union of sperm & egg) divides to form blastocysts
• The inner cell mass of the blastocyst generates the embryo.
• Pluripotent cells of the inner cell mass known as embryonic stem (ES)
cells can be induced to differentiate into cells of all 3 germ layers
• In the embryo pluripotent stem cells can:
• Asymmetrically divide to yield a residual stable pool of ES cells
• Generate populations that have progressively more restricted
developmental capacity, eventually generating stem cells that are
committed to just specific lineages.
Fundamentally there are only 2 varieties of stem cells
1. Embryonic stem cells (ES cells)
a. Are the most undifferentiated, present in the inner cell mass of the blastocyst, with
limitless cell renewal capacity & can give rise to every cell in the body & are thus said to
be Totipotent
b. ES cells can be maintained for extended periods without differentiating; thereafter
can form specialized cells of all 3 germ cell layers when appropriately stimulated
2. Tissue stem cells (Adult stem cells)
a. Found in intimate association with differentiated cells of a given tissue.
b. Are normally protected within specialized tissue micro-environments called stem cell
niches like in the bone marrow where hematopoietic stem cells congregate in a
perivascular niche.
Other niches
a. Bulge region of hair follicles
b. Limbus of the cornea
c. Crypts of the GIT
d. Canals of Hering in the liver
e. Subventricular zone in the brain.
Soluble factors & other cells within the niches keep the stem cells quiescent until
there is a need for expansion & differentiation of the precursor pool
c. Adult stem cells have a limited repertoire of differentiated cells that they can
generate.
ADULT STEM CELLS
• Can maintain tissues with high (Skin & GIT) or low ( endothelium) cell turnover
• Adult stem cells in any given tissue can usually only produce cells that are normal
constituents of that tissue.
• Hematopoietic stem cells continuously replenish all the cellular elements of the
blood as they are consumed.
• Can be isolated directly from bone marrow & peripheral blood after administration
of certain colony stimulating factors (CSF) that induce their release from bone
marrow niches.
• Can be purified to virtual homogeneity based on cell surface markers, clinically
used to repopulate marrows depleted after chemotherapy (Leukemia) or to
provide normal precursors to correct various blood cell defects (Sickle cell disease)
• Besides hematopoietic stem cells, the bone marrow & especially fat
contains a population of mesenchymal stem cells (Multipotent cells
that can differentiate into a variety of stromal cells including
chondrocytes (cartilage), osteocytes (bone), adipocytes (fat) &
myocytes (muscle).
• Because these cells can be expanded to large numbers, they
represent a potential means of manufacturing the stromal scaffolding
needed for tissue regeneration.
REGENERATIVE MEDICINE
• Defined as the identification, isolation, expansion & transplant of stem
cells
• Differentiated progeny of ES or adult stem cells can be used to repopulate
damaged tissues or to construct entire organs for replacement.
• Especially therapeutic opportunities for restoring damaged tissues that
have low intrinsic regenerative capacity (Myocardium from myocardial
infarction or neurons after a stroke.
• It is now possible to generate pluripotential cells resembling ES cells
derived from the patient into whom they will be implanted.
• Some genes have been identified whose products can remarkably
reprogram somatic cells to achieve the “stem-ness” of ES cells. When
such genes are introduced into fully differentiated cells like
fibroblasts, induced pluripotent stem cells (iPS cells) are generated
• Because the cells are derived from the patient, their differentiated
progeny can be engrafted without eliciting an immunologically
mediated rejection reaction that would occur if the differentiated
cells were derived from ES cells obtained from another donor.
• The production of induced pluripotent stem cells (iPS cells).
• Genes that confer stem cell properties are introduced into a patient’s
differentiated cells giving rise to stem cells that can be induced to
differentiate into various lineages