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NormalTissue

Tumor inductionCarcinogenGEMM

GENETIC BACKGROUND

Heterogeneous mousepopulation

BenignTumor

InvasiveTumor

Metastasis Therapy Relapse

Exposure toCarcinogens, promoters

Modelling Human Cancer in the Mouse

Angiogenesis, stromal cellsImmune systemInflammation

Somatic mutations

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Chemical carcinogen models

Radiation models

Lifestyle models

Susceptibility

Summary

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Initiation

Promotion

Epidermal Cell

Initiated Cell

Benign Tumor

MetastaticTumor

MalignantTumor

Progression

DMBA TPA papilloma

Multistage skin carcinogenesis

Hras mutMutant Ras amplificationLoss of wt Ras

Invasion

Epithelial-mesenchymalTransition (EMT)

Metastasis

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HRAS NRAS KRAS

Breast 1/200 (~0%) 0/147 (0%) 11/270 (4%)

Large Intestine 2/501 (~0%) 11/404 (3%) 4283/13564 (32%)

Lung 7/1355 (1%) 21/1868 (1%) 1568/8427 (19%)

Melanoma 89/1585 (6%) 491/2702 (18%) 31/1288 (2%)

Pancreas 0/191 (0%) 4/215 (2%) 2637/4388 (60%)

Salivary Gland 22/111 (20%) 0/30 (0%) 1/45 (2%)

Ovary 0/94 (0%) 4/109 (4%) 209/1234 (17%)

Haematopoietic and lymphoid tissue

8/2345 (~0%) 756/6077 (12%) 204/3913 (8%)

Endometrium 4/293 (1%) 1/277 (~0%) 224/1548 (14%)

Ras Mutations in Human Cancer

Sanger Center COSMIC database (www.sanger.ac.uk/genetics/CGP/cosmic/)

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p c c c c c c c

normal

mutant

H-ras

CARCINOGEN-INDUCED H-RAS MUTATIONS IN SKIN TUMORS

Quintanilla et al

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H-ras and skin cancer

1. Skin tumors induced by the same carcinogen carry the same oncogene: H-ras

2. Skin tumors induced by DMBA have the same activating mutation in H-ras

3. Mutations occur early – at time of initiation

4. Amplification of mutant ras or loss of wild type ras occurs during progression

(circa 1982- 1986)

5. Tumours from F1 hybrid mice show preferential mutation of one parental allele

6. Some mouse strains are resistant to ras-initiated tumours.

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UrethaneDMBA

Lung Adenomas

90% K-ras mutations

Tissue-specific Ras mutations

Conclusions: H-ras is required for skin, and K-ras for lung tumors

Skin papillomas/carcinomas

90% H-ras mutations

TPA

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ATMAtaxia telangiectasia

BRCA1Familial breast and

ovarian cancerp53

Li-Fraumeni syndrome

NBS1-MRE11-RAD50Nijmegen breakage syndrome

Ataxia-telangiectasia-like disorderCDS1/CHK2

Li-Fraumeni syndrome

Cell-cycle checkpoints DNA repair

Double-stranded breaks in DNARadiation

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Mouse genetic models for radiation-induced cancers

4Gy

NO tumors

Kemp et al, p53 deficient mice are extremely susceptible to radiation-induced tumorigenesis. Nature Genetics, 8(1):66-69 (1994)

P53+/-

LymphomasSarcomas

4Gy

Delta P p53m/m

Solid tumors

OR

Fbxw7+/-

4Gy

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0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

0 10 20 30 40 50 60 70 80 90 100

Age (weeks)

Pro

po

rtio

n o

f S

urv

ivin

g

Spon 129/Sv (52)

Rad 129/Sv (73)

p53+/- mice are extremely susceptible to radiation-induced tumorigenesis

Kemp et al, Nat.Genet, 1994

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0

0.2

0.4

0.6

0.8

1

0 10 20 30 40 50 60

Age (weeks)

Pro

port

ion

of T

umor

-Fre

e

(Spretus x 129/Sv)F1 (70)

(B6 x 129/Sv)F1(95)

129/Sv (73)

Genetic background affects radiation-induced lymphoma susceptibility

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Gct4

Genetic changes in radiation-induced tumors

0

10

20

30

40

70

50

60

80

90

7 8 9 10 11 121 2 3 4 5 6 13 14 15 16 17 18 19

PctmHcf2Hcs7Pas8

Pas6

Sluc3

Pas1

Sluc4

Par1

Lts1

Par2

Pas3

Sluc1

Hcs4

Hcr1Hcs5

Hcs1Skts1

Skts2

Skts3

Skts5

Skts11

Skts10

Hcs2

Hcr2

Hcs3

Hcs6

Scc3

Scc1

Scc2

Ccs1 Scc4

Scc5

Ssic1Mom1

Ter

Tli1

Tlag1

Tlsr2

Tlsm1

Foc1

Esl1

Msmr1

Esl1

Fv1

Rmcf

Fv2

Fv4

Rfv3Rfv1,2

Ril3

Rvil1

Lyr1

Ril1

Rmv1,2,3

Pctr1

Pctr2

Pctr1

Rrs

Sluc6

Sluc7

Sluc8

Sluc9

Sluc10

Skts6Pas6

Sluc11Sluc12

Sluc13

Hcf1Sluc14

Msmr2

Papg1

Par4

Liver

Liver

Ots1

Rgv1

Lyr2

Psl1

Skts8

Skts7

Skts12

Skts9

Scc7Ccs2

Pas9

Gct1

Skts4

Scc8Pas4

Scc9

Pas5b(Pas5)

Par3

Gct2

Scc6

Pgct1

Pas7

Tgct1

X

Skts13

Gct3

Foc1

Lyr3

Rfv3

Ikaros

STK15: Mitotic checkpoint

Fbxw7/Cdc4

Notch1

Myc

P53-dependent

HipK2 P53-independent

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The Circadian Clock

Fu and Lee, Nature Reviews Cancer, 2003

Clock genes and cancer?

Female shift workers have increasedRisk of breast and/or colon cancer

Mouse knockouts of Per2Have increased colon tumors andlymphoma

A polymorphism in human PER3 isAssociated with breast cancer

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Ko, C. H. et al. Hum. Mol. Genet. 2006 15:R271-277R; doi:10.1093/hmg/ddl207

Control of the Mammalian Circadian Clock

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“GOOD PROGNOSIS” Breast cancer

Van’t Veer et al, 2002

NETWORK ANALYSIS OF HUMAN BREAST CANCER

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Aurora-A

Aurora-B

CyclinB

Blooms

Mitotic spindleprotein

Never-in mitosisGene a

cytokinesis

Mitotic spindle checkpoint

Anaphase spindleElongation 1

Mitotic-specificUbiquitin-conjugating

Spc25Kinetochore protein

UBE2T(Fanc)

PER3

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Per3Per3

Tumor suppressor genes on human 1p36 (mouse chr.4)

Chd5 Bagchi et alCell 2007Kif1b- Schlisio et al

Genes Dev. 2008

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0 5 10 15 200.0

0.2

0.4

0.6

0.8

1.0

Time (Years)

Dis

ea

se

Fre

e S

urv

iva

l

p= 0.0009

DFS at 10 years ± SE (%) 69 ± 3 vs. 56 ± 5

ALL PATIENTSN= 413

122

PER3 LOW

291

PER3 NORMAL/HIGH

39% RECURRENCE 27% RECURRENCE

P=0.013

LOW EXPRESSION OF PER3 IS ASSOCIATED WITHPOOR PROGNOSIS

Van de Vijver et al, NEJM 2002Chin et al Cancer Cell 2006

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0.0

0.2

0.4

0.6

0.8

1.0

0 5 10 15 20

Time (Years)

Pro

bab

ility

of

Dis

ease

Fre

e S

urv

iva

l

p= 0.0003

74 ± 3 vs. 54 ± 5

0.0

0.2

0.4

0.6

0.8

1.0

0 5 10 15 20

Time (Years)

Pro

bab

ility

of

Dis

ease

Fre

e S

urv

iva

l

p= 0.9

56 ± 5 vs. 57 ± 6

BUT ONLY IN ER-POSITIVE TUMORS

ER+ ER-

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Per3 null mice are susceptible to breast cancer

5 10 15 20 250.0

0.2

0.4

0.6

0.8

1.0

0

Time (Months)

Pro

babi

lity

of T

umor

Fre

e S

urvi

val

p= 0.003

B

0%

MMTV-neu per3

Genotype

05

10152025303540

WT (n=17) Het (n=33) Null (n=28)B

reas

t ca

nce

r in

cid

ence

(%

)

A

12%

36%p= 0.005

DMBA gavage

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Conclusions

PER3 is a tumor suppressor gene on 1p36

Deletion of human PER3 is associated with increasedRisk of recurrence in ER positive tumors

Loss of Per3 has a causal effect in increasing breast cancer in mice.

Make sure you get plenty of sleep…