Cancer Genetics

Chapter 23

http://www.labnews.co.uk/news_archive.php/761/5/visions-of-science-winners-chosen/
1 out of 2 men
1 out of 3 women
group of diseases characterized by cells that do not
respond to the normal controls on cell division

-caused by multiple
acquired somatic
mutations that occur
over time

-separate inherited
susceptibility in
some cases
one mutation
already in all cells
-tumor
localized benign
invasive malignant

What is cancer?
Lung tumor
http://science.nationalgeographic.com/science/enlarge/lung-tumor.html
http://science.education.nih.gov/supplements/nih1/cancer/activities/activity2_animations.htm
Lung tumor
http://science.nationalgeographic.com/science/enlarge/lung-tumor.html
What is cancer?
group of diseases characterized by cells that do not
respond to the normal controls on cell division

-caused by multiple
acquired somatic
mutations that occur
over time

-separate inherited
susceptibility in
some cases
one mutation
already in all cells

-tumor
localized benign
invasive malignant

Relies upon
signals from
other cells
Cancer cells
disconnect from
normal regulation
as part of multicellular
Organism
Renegade Cell
Tapping into early
developmental pathways
http://www.hhmi.org/biointeractive/media/angiogenesis-sm.mov
http://www.hhmi.org/biointeractive/media/angiogenesis-sm.wmv
per child = 10
-8
mutation rate
35 & 49 new germ-line mutations
in 1000 genomes project
Production of sperm and eggs
~23 cell divisions to
make each gamete
1 fertilized cell to ~10
14
somatic cells
in body
1,586 new mutations in single somatic cells
measured in 1000 genomes project

Mutation rate ~1x10
-10
per bp per cell division
so ~ 0.6 new mutations each time a cell divides in humans
Germ-line mutations
somatic mutations
Cancer normally results from somatic mutations
if have 10
10
cell divisions, some cell has
likely mutated one of each base pair in genome
NCI
Quickly proliferating cells- e.g. epithelial cells
Cells whose growth is controlled by hormones
Cells exposed to environmental carcinogens
Clonal Evolution of Tumors
Cancer begins with a mutation in a single cell
leads to abnormal cell replication
extra mutations occur in subsequent cells
cells carrying both take over since grow faster
new mutations occur that enhance proliferation
further
Cells evolve to grow faster and less impeded by
controls
Cancer-related genes
control the cell cycle

Mutations in these can disrupt
normal cell cycle controls
Cell Cycle
ensure DNA is replicated only once per cycle
ensure DNA is intact and cell is ready
ensure equal distribution of DNA to daughter cells








Checkpoints control progress to next stage
Controlled by cyclins and cyclin-dependent kinases

Cell cycle game:http://www.nobelprize.org/educational/medicine/2001/cellcycle.html
Mutated Genes that contribute to
Cancer
Genes associated with the regulation
of the cell cycle
stimulate growth and cell division
mutation makes it hyperactive or active at
inappropriate times
usually dominant, so just need one mutation
ONCOGENES/ PROTOONCOGENES

inhibit growth and cell division
mutation makes gene inactive
usually recessive, so need two mutations
TUMOR-SUPPRESSOR GENES
Oncogenes
mutation stimulates cell growth
Exist in normal cells as protooncogene
when mutated to oncogene, stimulates growth
Can test by inserting an oncogene into other
cells and see growth stimulation

1910 Peyton Rous
Rous sarcoma virus
v-src is a viral gene that can cause cancer when added to
cells
1975 Bishop and Varmus
found cellular proto-oncogene
c-src is an unmutated copy that functions normally
SRC- protooncogene
Biochim Biophys Acta 1602(2), M. Frame, Src in Cancer, 114-130, 2002
Oncogene version vSRC loses tail with Y527
Loses inhibitory part so always active
http://en.wikipedia.org/wiki/Apoptosis
Many proto oncogenes are involved in signaling
external stimulatory signal increase in cell growth
Cell-cell
contact
Stimulatory signals
G1/S
http://en.wikipedia.org/wiki/Apoptosis
Many proto oncogenes are involved in signaling
external stimulatory signal increase in cell growth
mutations - over express protein
-stick protein in on position
Cell-cell
contact
G1/S
RAS protein is a GTP/GDP switch like translation factors
Mutation that affects ability to hydrolyzed GTP sticks in ON position
Dominant acting

http://courses.washington.edu/conj/gprotein/monomericgp.htm
Mutations can stick signaling protein in an always ON position
Normal Myc function
Signals that regulate Myc
Cell contact
Growth factors
Cytokines

Differentiation of cell
Contact Inhibition

Seminars in Cancer Biology
Volume 16, Issue 4, August 2006, Pages 318-330
Cancerous Growth of Cell
Example: Myc/Max/Mad
-Activation of MYC by translocation
Translocation of Myc gene at
breakpoint onto one of three other
chromosomes
-fuses to immunoglobin promoter so
turns on myc in B-cells

Burkitt B-cell carcinoma
http://8e.devbio.com/article.php?ch=5&id=42
Ways Myc gene expression can be increased:
- Mutations in regulatory proteins and pathways e.g. NFAT
- Duplication of myc gene
c-myc gene
c-myc gene promoter
Strong promoter
on in white blood cells
Myc promoter
immunoglobin
promoter
promoter
Tumor Suppressor Genes
normally limit cell growth
Normal gene limits progression through cell cycle or
checkpoints unless certain conditions are met
Normally inhibits cancerous growth

Mutation of the gene removes this control and allows
faster or unhindered cell growth

examples
p53
rb
APC
BRCA1

Cell Cycle Regulation
Checkpoints
G
1
/S checkpoint
DNA damage
Environmental
suitability
external growth factors
G
2
/M checkpoint
centrosome/DNA duplication, DNA damage
Spindle checkpoint
Chromosome alignment in metaphase
http://www.cellsalive.com/cell_cycle.htm
http://en.wikipedia.org/wiki/Apoptosis
Cell-cell
contact
Apoptosis
cell death pathway
-fail safe if
something goes
wrong
Self
destruct
Tumor suppressor genes often act as
inhibitors that signaling overcomes,
or part of error recognition that halts
or triggers apoptosis
Familiar risk if inherit one defective allele,
then mutation just needs to hit the second allele to start road to cancer
Starts S phase
E2F is transcription
factor that turns on
growth-related genes

RB binds to E2F to prevent
the cell from moving into S
unless all of the cellular
components are available
and the cell is ready for
growth.


Mutation of RB allows cell
to immediately enter S
phase

RB- tumor suppressor gene
RB- tumor suppressor gene

Alfred Knudson (1971)
retinoblastoma

Unilateral
Retinoblastoma
Sporadic-not inherited
Bilateral
Retinoblastoma
Runs in families
tumor
two hits in a single cell
would happen only rarely;
results in a tumor in one eye
Single eye tumor (Unilateral)
No family history
Tumors in both eyes (Bilateral)
Usually runs in families
genetic
predisposition
because only a single
mutation is needed, the
likelihood of it occurring in
both eyes is increased
Many inherited bilateral retinoblastoma families have
deletion on Chromosome 13 13q14- includes RB gene
RB
RB
RB
G
1
/S checkpoint
Two important questions at this
checkpoint:
IS THE ENVIRONMENT PERFECT FOR
CELL DIVISION?
IS DNA DAMAGED IN ANY WAY?

BRCA1 & 2 and P53 detect damaged
DNA
BRCA1 and BRCA2
Associated with breast and ovarian cancer
inherited defective BRCA1 allele is associated
with only 5% of breast cancer cases
Heterozygote female has 80% risk of developing
breast cancer by age 80

links detection of DNA damage to control of
cell cycle G2/M
Part of a DNA repair complex
arrest division until damage is fixed

You tube cartoon: http://www.youtube.com/watch?v=G8kFs2lMdfY
Also- bulky blocks to DNA replication or transcription trigger ATM
Trends in Molecular Medicine
Volume 8, Issue 12, 1 December 2002, Pages 571-576
National Cancer Institute http://www.cancer.gov/cancertopics/pdq/genetics/breast-and-ovarian/HealthProfessional/page1
Missense
Cys61Gly
Arg814Trp
Ser1040Asn
Met1775Lys
Met1775Arg

Deletions
185delAG Ashkenazi Jews
1294del40
1675delA Norway
2800delAA
3121delA
3888delGA
4153delA Russia
4184del4
5085del19
Insertions
943ins10 Africa
1135insA Norway
3171ins5 Sweden
5382insC
5438insC
5677insA
Splice site variants
330A-G 5 splice
exon 5
Arg71Gly
BCRA1 Mutations linked to familial risk
http://www.nytimes.com/2007/09/16/health/16gene.html?pagewanted=all&_r=0
BRCA1 Previvor video: http://video.on.nytimes.com/video/2007/08/31/health/1194817106561/the-story-of-a-previvor.html
http://www.mun.ca/biology/desmid/brian/BIOL2060/BIOL2060-19/1939.jpg
P53
Guardian (or Tattletale)
of the genome




50% of all human cancers are associated
with defects in p53



If a cell has incurred DNA damage:
-turns on genes to repair the damage
-arrest division until damage is fixed
-initiate apoptosis (cell death) if not
fixed
-P53 mutations in red in DNA-binding residues- makes it act
as a dominant-negative . Unusual for a tumor suppressor gene
-Mutations in the promoter, frameshift, nonsense mutations eliminate function-
those act as typical recessive tumor-suppressor gene.
http://www.mun.ca/biology/desmid/brian/BIOL2060/BIOL2060-19/1939.jpg
P53
Guardian (or Tattletale)
of the genome
If can not bind, it can not
trigger arrest/apoptosis
Korean J Lab Med. 2008 Dec;28(6):493-497
Li-Fraumeni syndrome
One copy of p53 mutated so only inherits one functional copy

Dominant autosomal inheritance of early cancer in many different organs

P53 misense mutation 13203G>A Arg175His

APC - a tumor suppressor gene
controls pathway regulating cell-cell adhesion


Trends in Biochem Sci Volume 26, Issue 11,
Non-cancerous
cancer
Colon Epithelial cells
Clinical Cancer
Research July
15, 2006; 12
(14)
cadherin
-catenin
APC - a tumor suppressor gene
also in mitotic spindle checkpoint

checks on spindle attachment
Binds microtubles attached to
kinetochore of chromosomes

mutations lead
to aneuploidy
Nature Reviews Cancer 1, 55-67 (October 2001)
doi:10.1038/35094067
http://berkeley.edu/news/media/releases/2007/06/26_drugresistance.shtml
Karyotypes
http://www.nature.com/labinvest/journal/v80/n7/full/3780108a.html
Two different lung cancer cell lines
Changes in Other Regulatory Types
Increased chromosomal abnormalities
Aneuploidy, translocations, deletions

Changes in DNA methylation
Decreased or increased methylation of CpG
e.g. APAF1 (required for apoptosis) is hypermethylated in leukemia

Changes in miRNAs up regulate other protooncogenes or turn off other
tumor suppressor genes
Database: http://www.mir2disease.org/
e.g. mir15.1-16.1 cluster on chrom 13q14- deletion linked to B cell and
prostate cancer

Selection for changes that produce faster growth:
~Delete or silence more tumor suppressor genes

~Multiply copies or enhance expression of
-Oncogenes
-Genes for Angiogenesis (blood vessel growth)
-Genes for Cell Migration- allows metastasis



Progressive mutations
accumulate to allow:
Uncontrolled cell growth
Genomic instability
Immortality (telomerase activated)
Angiogenesis- recruiting blood vessels
Invasiveness (metastasis)
Personalized tumor genetics:
What genes have changed in MY tumor?

DNA chip- detect genes that are duplicated or lost in tumor cells
Transcript DNA chip- measure mRNA transcripts of genes- overexpressed or silenced
Breast Cancer in tumor cells
Tumors that overexpress Her2 (human epidermal growth factor receptor 2 tyrosine kinase)?
Can use drugs trastuzumab (Herceptin) and lapatinib (Tykerb) Her2 & EGFR
induce RAS
Tumors that overexpress epidermal growth factor receptor (EGFR)? & angiogenesis
Can use drugs cetuximab (Erbitux) and erlotinib (Tarceva)

Tumor overexpresses estrogen receptor (ER) ? Progesterone Receptor (PR)?
If yes, can use antiestrogens tamoxifen (Valodex) an estrogen antagonist,
if no, these will not work

Tumors caused by BRCA mutations?
Can use drugs PARP inhibitors (poly ADP ribose polymerase) e.g. Veliparib
Relevant Genes in whole body, not just tumor:
Mutations in CYP2D6 that reduce activity?
Do not prescribe tamoxifen- drug can not be activated in body





Colorectal Cancer
arises in cells lining the colon and
rectum
statistics
~150,000 new cases/yr
Progressive mutations
Familial adenomatous polyposis coli
-Inherits one defective copy APC
autosomal (5q22.2) dominant
-hundreds of mini polyps
if not removed, one usually becomes malignant

Polyp- benign
Sporadic tumors
75% have APC mutated

Familial APC- one copy
already mutated
70% tumors
have P53 mutated
Environmental Risks of Cancer
mutagenic conditions increase chance of mutation
Chemical exposure
Air pollution, food, environmental toxins
(smoking)
Diet
- Carcinogens (plant and microbial toxins), fat
Radiation exposure
Sunlight, radon
Infectious agent
Viruses: HPV, SV40, Hepatitis B, Epstein-Barr
virus, Retrovirus
Medical treatments
estrogen replacement therapy
http://en.wikipedia.org/wiki/Human_papillomavirus
Human Papilloma Virus
Vaccine- Gardasil targets 6,11,16 & 18
HeLa cancer cell line
from Henrietta Lacks cervical
cancer 1951
HPV infection
http://www.itmhrt.ca/course2.html
Human Papilloma Virus (Cervical Cancer strains)
- 2 genes E6 & E7 interact with tumor suppressors to assure viral
take over of cell cycle
http://www.cancer-therapy.org/CT3B/HTML/46.%20Jo%20and%20Kim,%20419-434.html
E6- binds to P53 E7- binds to RB
Hepatitis B
Aflatoxin- mutagen
Aspergillus
Liver Cancer
China has world cases
http://www.niehs.nih.gov/health/impacts/aflatoxin.cfm
Burkitt's Lymphoma

Epstein-Barr Virus EBV
95% of US adults have been infected
if not infected until adolescent or later- develop mono

In Africa, early EBV infection + recurrent childhood malaria
Malarial Infection
+
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Xu PNAS 2013 vol. 110 no. 26 10759