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Dong-Young Noh • Wonshik Han • Masakazu Toi
Editors

Translational Research
in Breast Cancer
Editors
Dong-Young Noh Wonshik Han
Department of Surgery, College of College of Medicine
Medicine Seoul National University College of Medicine
Seoul National University Seoul, Korea (Republic of)
Seoul, South Korea

Masakazu Toi
Graduate School of Medicine
Kyoto University Graduate School of
Medicine
Kyoto, Japan

ISSN 0065-2598 ISSN 2214-8019 (electronic)

Advances in Experimental Medicine and Biology
ISBN 978-981-32-9619-0 ISBN 978-981-32-9620-6 (eBook)
https://doi.org/10.1007/978-981-32-9620-6

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Acknowledgement

Those listed below contributed to this publication by organizing the contents and
authors of textbooks, collecting and arranging manuscripts as assistant editors.

Department of Surgery, Seoul National Hyeong-Gon Moon

University College of Medicine, Han-Byoel Lee
Seoul, Republic of Korea Hong-Kyu Kim

v
Contents

Part I Introduction
1 Translational Research in Surgical Oncology: Introduction
and My Own Experience as a Surgeon-Scientist . . . . . . . . . . . . . . . 3
Dong-Young Noh

Part II Breast Cancer Biology and Cell Signaling Pathways

2 Phospholipase Signaling in Breast Cancer . . . . . . . . . . . . . . . . . . . . 23
Yu Jin Lee, Kyeong Jin Shin, Hyun-Jun Jang, Dong-Young Noh,
Sung Ho Ryu, and Pann-Ghill Suh
3 HER2 Signaling in Breast Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . 53
Incheol Shin
4 Ras Signaling in Breast Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Aree Moon
5 Epigenetic Regulation in Breast Cancer . . . . . . . . . . . . . . . . . . . . . 103
Hye Jin Nam and Sung Hee Baek
6 Role of tRNAs in Breast Cancer Regulation . . . . . . . . . . . . . . . . . . 121
Nam Hoon Kwon, Jin Young Lee, and Sunghoon Kim
7 Fusion Genes in Breast Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
Jisun Kim and Wonshik Han
8 DNA Damage Repair Inhibitor for Breast Cancer Treatment . . . . . 159
Ahrum Min, Kyung-Hun Lee, and Seock-Ah Im

Part III Cancer Stem Cell and Tumor Heterogeneity in Breast Cancer
9 Breast Cancer Metastasis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Mi Young Kim

vii
viii Contents

10 Single Cell Genomics for Tumor Heterogeneity . . . . . . . . . . . . . . . . 205

Hae-Ock Lee and Woong-Yang Park
11 Advances in Tumor Sampling and Sequencing in Breast Cancer
and their Application in Precision Diagnostics and Therapeutics . . . 215
Amos Chungwon Lee, Han-Byoel Lee, Huiran Yeom, Seo Woo Song,
Su Deok Kim, Ahyoun Choi, Sumin Lee, Yongju Lee, Wonshik Han,
and Sunghoon Kwon
12 Cancer Stem Cells in the Immune Microenvironment . . . . . . . . . . . 245
Dong-Sup Lee and Keunhee Oh
13 Theranostics for Breast Cancer Stem Cells . . . . . . . . . . . . . . . . . . . 267
Woo Kyung Moon and Hoe Suk Kim
14 Patient-Derived Xenograft Models in Breast Cancer Research . . . . 283
Deukchae Na and Hyeong-Gon Moon

Part IV Biomarkers for Precision Medicine in Breast Cancer Patients

15 Proteomic Interrogation in Cancer Biomarker . . . . . . . . . . . . . . . . 305
Un-Beom Kang
16 Next-Generation Sequencing-Based Biomarkers in Breast
Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 323
Wonshik Han and Woosung Lim
17 Liquid Biopsy in Breast Cancer: Circulating Tumor Cells and
Circulating Tumor DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 337
Tae-Kyung Yoo
18 Current Biomarkers for Precision Medicine in Breast Cancer . . . . 363
Soo kyung Ahn and So-Youn Jung
19 The Potential Predictors in Chemotherapy Sensitivity . . . . . . . . . . . 381
Eun-Kyu Kim and Hee-Chul Shin
20 Hormone Resistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 391
Jonghan Yu

Part V Managing High Risk Women

21 High-Risk Population Based on BC Risk Factors . . . . . . . . . . . . . . 405
Sue K. Park and Keun-Young Yoo
22 Breast Cancer-Related Low Penetrance Genes . . . . . . . . . . . . . . . . 419
Daehee Kang and Ji-Yeob Choi
23 Rare Coding Variants Associated with Breast Cancer . . . . . . . . . . . 435
Mi-Ryung Han
Contents ix

24 Multigene Panel Testing for Hereditary Cancer and Genetic

Counseling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 455
Eun-Shin Lee, Jongjin Kim, and Wonshik Han
25 BRCA and Breast Cancer-Related High-Penetrance Genes . . . . . . . 473
Sang-Ah Han and Sung-Won Kim

Part VI Next Generation Clinical Research

26 Clinical Databases for Breast Cancer Research . . . . . . . . . . . . . . . . 493
Ki-Tae Hwang
27 Care for Breast Cancer Survivors . . . . . . . . . . . . . . . . . . . . . . . . . . 511
Su Min Jeong and Sang Min Park
28 Considerations in Oncoplastic Surgery . . . . . . . . . . . . . . . . . . . . . . 525
Min Kyoon Kim and Jaihong Han
29 Diet Before and After Breast Cancer . . . . . . . . . . . . . . . . . . . . . . . . 545
Jung Eun Lee
30 Current Trends in and Indications for Endoscopy-Assisted Breast
Surgery for Breast Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 567
Hyukjai Shin
31 Minimal Invasive and Individualizing Management of the Axillary
Nodes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 591
Jun Won Min and Jihyoung Cho
32 Malignant Phyllodes of Breast . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 601
Cha Kyong Yom
33 Next-Generation Clinical Trials and Research with Successful
Collaborations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 613
Masakazu Toi and Ravi Velaga

Part VII Epilogue

34 Transformation of the Patient and Society: A Patient Survivors’
Group and Breast Cancer Awareness Campaign . . . . . . . . . . . . . . . 625
Dong-Young Noh
 Part I
Introduction
Chapter 1
Translational Research in Surgical
Oncology: Introduction and My Own
Experience as a Surgeon-Scientist

Dong-Young Noh

Abstract Translational research is possible when scientists have broad knowledge

of not only basic research, but also clinical science, which is acquired via experience
in patient care. These requirements cannot always be met by one individual, and,
hence, collaboration between suitably qualified individuals is the key for the pro-
gress of translational research. However, it is vital that translational research is
conducted by an investigator who has knowledge about all fields. I could be a
good conductor in that sense, because as an oncology surgeon, I have considerable
experience in working with patients; in addition, I have a background in biochem-
istry and have started my basic research laboratory. Thus, I can use these qualifica-
tions to my advantage to build a tissue bank as the first step, and initiate small-scale
experiments such as estimating the DNA or protein levels in specific tissues or blood
samples. Once I successfully launch good research products and publish in peer-
reviewed journals, I intend to build a large research group focusing on large-scale
studies on single nucleotide polymorphisms and proteomics. These translational
approaches can overcome several unsolved clinical problems. Many of my research
products, for example, patents and new techniques such as Mastocheck@, are
designed for improving the clinical outcomes in patients.

Keywords Breast cancer · Surgical oncology · Translational research ·

Bioinformatics · Genomics · Proteomics

D.-Y. Noh (*)

Department of Surgery, College of Medicine, Seoul National University, Seoul, South Korea
Gangnam CHA Medical Center, Seoul, South Korea
e-mail: [email protected]; [email protected]

© Springer Nature Singapore Pte Ltd. 2021 3

D.-Y. Noh et al. (eds.), Translational Research in Breast Cancer, Advances in
Experimental Medicine and Biology 1187,
https://doi.org/10.1007/978-981-32-9620-6_1
4 D.-Y. Noh

1.1 Introduction

Modern medical service has improved considerably due to advancements in medical

science, which has enabled curation of scientific evidence and has allowed physi-
cian’s access to information regarding diseases. In particular, translational research
has a vital role in bridging basic science and patient concerns. One advantage of
being an oncology surgeon specialized in breast cancer is the easy access to normal
or diseased human tissues, along with basic knowledge of human anatomy. I started
building my own human tissue bank in 1990, which now includes both cancer tissue
and adjacent normal tissues. All the tissues are frozen in liquid nitrogen and stored in

70  C or
20  C freezers. Tissue banking is important as a resource for conducting
research using diseased organs and normal tissues. The process of establishing a
tissue bank starts from obtaining permission from Institutional Review Board (IRB)
and patients or from normal healthy individuals. The term “translational research”
had not been coined at the time when I initiated my tissue bank; however, a bed to
bench, bench to bed concept existed. Nonetheless, this tissue bank formed an
important resource for successful translational research.
Combining clinical practice and laboratory work was not easy for a clinician;
however, once I overcame the hurdles, it turned out to be the most appropriate way
of conducting translational research. In the beginning, I was able to start with a
technician and rent a small part of a bench in a biochemistry laboratory owned by my
colleague. My research efforts and small achievements led to the growth and
development of my own laboratory. My postgraduate medical students and gradu-
ates (PhD) from basic research laboratories were the key personnel who developed
ideas and conducted research in the field of translational medicine.
My postgraduate study on biochemistry as the major subject formed the basis of
my translational research. My thesis was on “Purification of membranous 5’ nucle-
otides.” and “Enzyme immunoassay of a-fetoprotein using monoclonal antibodies.”
These studies performed by clinicians were not common in the 1980s, but are now
available in the MD-PhD courses. After PhD, I spent two years in a biochemistry
laboratory in Building 3 at the National Institute of Health (NIH), Bethesda, Mary-
land, USA, as a Fogarty international postdoctoral fellow. I consider myself lucky to
be trained in both basic science and clinical practice in oncology.
During my term as a postdoctoral fellow at NIH, I concentrated only on labora-
tory work, without having to deal with patients or clinical work. Thus, I was able to
dedicate my time completely to basic science and worked toward developing my
project. I also gained the ability to design and troubleshoot my own research.
Translational research was originally defined as follows: To improve human health, scien-
tific discoveries must be translated into practical applications. Such discoveries typically
begin at “the bench” with basic research then progress to the clinical level, or the patient’s
“bedside.”
Source: National Cancer Institute, National Institutes of Health.

Translation research has a broader meaning and has been extended to computer
and cyberspace research at the bench to the bedside. Finally, translation is moving
1 Translational Research in Surgical Oncology: Introduction and My Own Experience. . . 5

from a disconnected unidirectional approach to an engaged bidirectional partner

approach between research laboratories to bedside, and from the bedside to the
community. Translational science encompasses many research areas involving
human, animal, organ, tissue, and cell line models. It also requires establishment
of networks between community and industry. All these components should collab-
orate to build good communication and feedback. This concept was also built by
myself, when I started the tissue bank and my laboratory as a surgeon scientist,
which was before the term “translational research” was introduced worldwide. I also
organized a group of patient survivors in 2000, and at the same time, I started a pink
ribbon campaign on the streets of Korea. All these activities were well organized and
has led to many scientific articles and social products. The first clinical aspect to
consider was creation of a patient database. The data should be of good quality with
standardized terms and each valuable should be as numerical as possible. Later we
built a web-based database for all breast cancer patients who were operated and also
followed the condition of the patients over time.
With time, the Laboratory of Breast Cancer Biology (LBCB) has transformed into
a perfect translational research platform. On the clinical side, tissue and blood
banking are performed at the operating room, clinical database is created using
data from clinics, and all pathological data collection and tissue banking were
performed at pathology laboratories; data from bioinformatics, sequencing, proteo-
mics, and other engineering experiments are obtained from collaborators, and
functional studies, animal experiments, and tissue and blood processing are
performed at the Medical College of LBCB’s Cancer Research Institute.
I started by establishing a cancer cell culture system as I was interested in cancer
stem cell biology. Thus, I successfully established a sphere culture system and was
able to generate my own cancer cell lines named SBCC1, 2, and 3, which are of
epithelial origin, and NDY of mesenchymal origin. These are all mammospheres
with different characteristics, expressing the epithelial marker EpCAM, with the
exception of NDY, which has sarcomatous characteristics such as rapidly growing
sarcospheres. All these cells grow as serial cultures and can also be transplanted in
NOD/SCID mice [1, 2].
Since 2001, we are participating in the Genomic Research Center for lung and
breast cancers, sponsored by the Ministry of Health and Welfare, Korea. We are
continuing our genomic studies, including single nucleotide polymorphism (SNP)
analysis and cDNA microarrays at the Genomic Research Center. Later in 2007, we
organized a group named Translational Research Organization for Cancer
(TROICA) for collaborative translational research. TROICA has enabled targeted
studies such as biomarker discovery, mining of prognostic predictors, and targeted
drug discovery. In addition, we were able to expand our basic research area not only
to proteomics, but also to genomics and aptamer development by collaborating with
the best scientists in each area in the country [3–5].
Networking between groups and individuals with the same purpose and aims is
interesting and scientifically satisfying. I aim to form a competent research group in
which the members enjoy their research and can share their experiences and ideas
regarding research in particular and life in general. These are the features of
6 D.-Y. Noh

translational research. In addition, I wish to transfer this legacy of combining basic

and clinical research to my junior faculties and postgraduate students.

1.1.1 Genomics

I have generated numerous publications from translational research in the field of

genomics related to SNPs, which are variations of single DNA building blocks
called nucleotides in genes. For example, conversion of nucleotide C to A is a
SNP. They occur once in every 300 nucleotides on average and are considered the
most common type of genetic variation. Most SNPs are benign, although some may
contribute to serious conditions such as breast cancer.
SNPs can be categorized into different subgroups similar to a pedigree (Fig. 1.1).
Those that fall in the coding regions are of two types: synonymous SNP and
non-synonymous SNP. Synonymous SNPs result in different codons, which encode
the same amino acid. Hence, synonymous SNPs are ineffective, as the building
blocks for proteins remain unchanged. However, missense or nonsense mutations
are formed when the codon and the amino acid it encodes change.
Studies for identifying the most common non-synonymous genetic variants that
are susceptible to breast cancer are limited [6]. A study showed that a novel SNP,
rs1053338(K264R) in ATXN7 at locus 3p21, is associated with susceptibility to
breast cancer. AKAP9-rs6964587 was also found to be a marker of a breast cancer
risk at 7q21 [7]. Both SNPs are susceptible to estrogen receptor (ER)-positive and
ER-negative disease [7]. Another locus, 2q35 rs 13387042, shows strong evidence
of association between rs13387042 and breast cancer in Caucasian women. This
SNP is also associated with both ER-positive and ER-negative breast cancer in
European women [8]. Another SNP from the same locus 2q35 was scrutinized. By
genotyping 276 SNPs using the 1000 Genomes Project data, the best functional
candidate, rs4442975, was found to be associated with ER+ among Europeans.

TYPES OF SNPs

Non-coding region Coding region

Synonymous Non- Synonymous

Missense Nonsense

Fig. 1.1 Breast cancer exhibits consistent genetic variation

1 Translational Research in Surgical Oncology: Introduction and My Own Experience. . . 7

Evidence shows that the g-allele increases breast cancer susceptibility via down-
regulation of IGFBP5, which is known to play a significant role in breast cancer
biology [9]. Genome-wide association studies (GWAS) has revealed that SNP
rs889312 in the 5q11.2 locus is associated with breast cancer risk in European
women. Functional analysis indicated that the cancer risk alleles of four candidates
(rs74345699, rs62355900, rs16886397, and rs17432750) increased MAP 3K1 tran-
scriptional activity. Cancer risk alleles act to increase MAP 3K1 expression in vivo
and might promote breast cancer cell survival [10]. Out of the 227,876 SNPs that
were estimated to correlate with 77% of the known common SNPs in Europeans, five
novel independent loci signaled strong and consistent evidence of association with
breast cancer. Four of these contain causative genes (FGFR2, TNRC9, MAP 3K1,
and LSP1). A second stage of the same research indicated that more SNPs can act as
susceptibility alleles [11].
Studies have been performed to locate the single nucleotide variation that can be a
critical factor for either inhibiting or accelerating tumor cell growth in breast cancer.
Certain types of SNPs can be found to be significantly associated with the overall
survival of patients due to their differential sensitivities toward certain drugs. Further
studies on these lines will ensure better outcomes for patients with breast cancer.
Initially, we hypothesized that if SNP can affect breast cancer development, it can
also play a role during disease progression and may change clinicopathological
features. We have observed that certain variants of CYP1A1 and CYP1B1 were
related with onset at younger age, and that a certain haplotype of BRCA1 showed less
ER negativity and another was associated more with lymph node-negative
phenotype [12].
Certain SNPs, for example those in HER-2, can affect tumor aggressiveness or
response to therapy, and, as a result, clinical outcome. In this study, the haplotypes
were not related with the risk of breast cancer; however, the most common haplotype
1 was associated with 1.5-times more frequent expression of HER-2 and showed
poorer prognosis than other haplotypes [13]. We have published 23 papers regarding
these SNP association studies in peer-reviewed journals.
The studies in LBCB regarding identification of SNPs for early detection of breast
cancer can be summarized as follows:
• Haplotype analysis of HER-2 polymorphism.
• Correlation between polymorphisms in DNA repair genes and susceptibility to
breast cancer occurrence using SNP chip.
• Breast cancer susceptibility of innate immunity- and non-Hodgkin’s lymphoma–
related genes.
• CASP8 polymorphism and breast cancer risk: A common coding variant in
CASP8 is associated with breast cancer risk.
8 D.-Y. Noh

1.2 Comparative Genomic Hybridization (CGH) Array

for Prognosticators

• Detecting prognostic factors in ER-positive breast cancer treated with tamoxifen

using the CGH array.
• Discovery of candidate clones associated with breast cancer systemic recurrence
using the CGH array.

1.2.1 Expression chip

• Investigation of differentially expressed genes and proteins during anoikis using

the breast cancer cell line MCF-7.

1.2.2 Immunohistochemistry (IHC)

• Utility of Ki-67 for predicting distant metastasis in node-negative breast cancer.

Among the other examples showing how research on SNPs can translate the
discoveries to the clinic, we investigated the correlation between significant SNPs
in DNA repair genes in breast cancer samples and breast cancer occurrence. We
evaluated the genetic polymorphisms (384 SNPs) in 38 DNA repair genes in a
hospital-based case-control study 480:480). The results were translated and patented
as breast cancer risk diagnosis SNP chip [14, 15]. Table 1.1 shows the results of
analysis of clone with gain or loss observed in more than 50% of the 77 samples in
the study [16]. For the clones selected in the analysis, a literature search, such as
NCBI and PubMed, confirmed their association with cancer and finally selected
eight candidate genes (Table 1.2) [16]. For the development of prognosticators after
treatment of breast cancer, we attempted to identify candidate clones associated with
breast cancer systemic recurrence using the CGH array and 31 pairs of breast cancer
patients matched for clinicopathological characteristics of recurrence cases and
recurrence-free cases after standard treatment [16] (Fig. 1.2).
Another interesting algorithm for predicting distant recurrence involved the use
of clinicopathological multimarkers and a decision tree. We developed a decision
tree for predicting prognosis from the 328 points of lymph node-negative breast
cancer patients and 38 recurrences using clinicopathological characteristics such as
age, tumor size, grade, and ER, PR, p53, c-erbB-2, and Ki-67 levels after adjuvant
treatments. The results were remarkably applicable (Fig. 1.3) [17].
1 Translational Research in Surgical Oncology: Introduction and My Own Experience. . . 9

Table 1.1 Common regions showing gain or loss in more tnan 50% of all 77 samples
Clone Number
Region No. Cytoband Start (kb) End (kb) (%) Cancer related genes
Gain-1 c5784 1p36.33 552,910 563,807 75 (97.4)
Gain-2 c5242 8q24.3 145,647,141 145,761,879 46 (59.7) CYHR1, KIFC2,
FOXH1, PPP1R16A,
GPT, MFSD3,
RECQL4, LRRC14,
LRRC24,
MGC70857,
KIAA1688
Gain-3 c4824 8q24.13 126,947,484 127,030,285 42(54.5)
Gain-4 c1394 8q23.3 116,937,688 117,027,644 41 (53.2)
Gain-5 c1437 8q24.12 119,396,534 119,465,174 41 (53.2) SAMD12
Gain-6 c1433 8q24.21 131,335,147 131,416,013 41 (53.2) DDEF1, DDEF1IT1
Gain-7 c2733 20q13.33 61,387,393 61,535,237 39 (50.6) ARFGAP1,
COL20A1,
CHRNA4, KCNQ2
Loss-1 c5256 8p23.1 7,323,700 7,428,919 57 (74.0) DEFB106B,
DEFB105B,
DEFB107B,
LOC645489,
FAM90A6P,
FAM90A7.
LOC729339,
FAM90A22,
FAM90A23
Loss-2 c4589 8p23.1 7,334,384 7,420,885 56 (72.7) DEFB105B,
DEFB107B,
LOC645489,
FAM90A6P,
FAM90A7,
LOC729339,
FAM90A22
Loss-3 c5126 8p23.1 7,647,665 7,716,751 55 (71.4) FAM90A19,
LOC729394,
FAM90A9.
FAM90A10,
DEFB107A
Loss-4 c5189 10q11.22 46,320,705 46,408,357 43 (55.8) RHEBP1, SYT15
Loss-5 c710 14q32.33 105,821,330 105,907,464 39 (50.6) IGHVIII-25-1,
IGHV2–26, IGHVIII-
26-1. IGHVII-26-2,
IGHV7–27,
IGHV4–28, IGHVII-
28-1, IGHV3–29
kb kilobase