12BraytonCancerPhenoModel

Mouse Cancer Models & Phenotypes  Cory Brayton, D.V.M. Diplomate, A.C.L.A.M., A.C.V.P. Director, Phenotyping Core Associate Professor, Molecular and Comparative Pathobiology Johns Hopkins University, School of Medicine Baltimore, MD 21205 TEL: 410 502 3050 -- FAX: 443-287-2954 [email protected]

http://www.hopkinsmedicine.org/mcp/PHENOCORE/

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Aims

Relevance to you ?

1. Learn more about our model mice to

Do you work with – Spontaneous mouse cancer models? – Induced cancer models?

help our cancer modeling  Nature & Nurture impact phenotypes in

• Carcinogens (potential carcinogens)? • GEM Genetically engineered mice ?

ALL experimental models.

2. Pathology can help our Research.  Confirm & characterize expected

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model phenotypes & results.  Diagnose unexpected phenotypes or results.

– Xenografts? – Acute or chronic studies? Do you use – Immunodeficient mice ? – Antibiotics ? 4 – BJC or ARRIVE or NAS guidelines?

Mouse Cancer Quiz

Mouse Cancer Quiz cont.

NATURE Match the cancer phenotype to the strain 1. Thymic Lymphoma 2. B cell Lymphoma 3. Plasma cell tumors (myeloma) (Harderian,

myoepithelioma, Rhabdomyosarcoma etc )

4. Lung tumors 5. Mammary tumors 6. Mammary Hyperplasia,

EMT with pituitary tumors

7. Liver tumor 8. Sarcoma, lymphoma, NET 9. Skin tumors, erythroleukemia

10.Cancer resistant

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a) A b) AKR, NODscid c) BALB/c d) e) f) g)

C3H, GR C57BL/6 DBA/2 FVB/N

h) SJL/J i) Tg.Ac j) Trp53 Tm

NURTURE What environmental or husbandry factors have been shown to affect tumor onset, incidence or responses to therapy?

a. b. c. d. e. f. g. h.

Antimicrobials Bedding Carcinogens Diet restriction Diet type Infectious agents Housing density Temperature

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Symbols – official & not so much

Discussion plan I. Cancer Models - Ideal VS Real II. Mouse Cancer phenotypes/models 1. Spontaneous: • •

Terminology Comparison to human tumors

2. Induced Carcinogens •

MORE !

 Famous mice

 Carcinogen  Radiation hazard  Mutagenicity  Reproductive

& strains

FASTER !

Toxicity

3. Induced GEM • 7

 Respiratory

examples of interfering phenotypes 

Ideal model of human cancer Relevant ! – Genetic / molecular – Biology / phenotypes

 Practical (time & $,€,₤) – Fast ($,€,₤) • Progression • Response to Rx • But not too fast (die too soon)

• What it looks Like & • How it behaves Histomorphology,

• Hi penetrance

– Primary Phenotype($,€,₤)

Metastasis, Response to Therapy

 Bloodborne pathogens (red)

genetically engineered mice

 Target Organ Toxicity

 Aspiration Toxicity

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Mouse Cancer Models

• i.e. not killed by anything else

Wait .. (Spontaneous)

Carcinogens

Genetic Engineering

– Cheap($,€,₤) • Not likely

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Xenografts

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Rodent Cancer Classification MMHCC – http://emice.nci.nih.gov/

Advantages +

Strategy

– Reproducible ($,€,₤)

Sites, distributions Progression,

 GEM

Sensitizer

what you want? When / where you want it ?

4. Xenografts •

 Biohazard

• Identify carcinogens • Identify treatment • Strain Environment influences • • • •

More /faster tumors identify carcinogens Identify treatments Strain Environment influences

Disadvantages

• Useful? Tumor types • Strain Environment influences

• Molecular mechanisms • Model behavior & response to treatment • Strain Environment influences

• Lethal or unexpected phenotypes • Expressed everywhere • Strain Environment influences

• Identify treatments

• Relevance of ‘cloned’ tumors • Infections in Immunodeficient • Strain Environment influences

• Strain Environment influences

Rodent Cancer Classification MMHCC – http://emice.nci.nih.gov/

• mouse models of human cancer consortium • Consensus papers  hematopoietic liver mammary prostate intestine ….

• GEM oriented – molecular mechanisms

INHAND - http://www.toxpath.org/nomen/

INHAND - http://www.toxpath.org/nomen/ • International Harmonization of Nomenclature and Diagnostic Criteria in Toxicologic Pathology

PATHBASE – http://www.pathbase.net/

• RENI Registry Nomenclature Information System

PATHBASE – http://www.pathbase.net/ 11

• Mouse GEM pathology ontology • NOT just cancer

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-

• Too slow ? Lo penetrance? • Not so human • Strain Environment influences

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Spontaneous MOUSE Tumors (Phenotypes to expect) Tumor

Hematopoietic Lymphoma, HS

Spontaneous MOUSE Tumors (Phenotypes to expect)

Sex

Strain predisposition

F>M

AKR, SJL/J, NODscid (C57BL, etc)

Lung

Mice definitely get other tumors.. Pituitary, Harderian, myoepithelioma, adrenal, thyroid, skin …

A (FVB, 129, etc)

– WNL (within normal limits) for your

Mammary

F>>M

C3H (BALB etc)

Liver

M>>F

C3H, CBA, (B6C3 …)

(aging) population? Or – Important • Expected/desired  model phenotype or

 What strains are famous for what tumors?

treatment related result (phenotype) ?

– Could that be useful to your tumor model ? 

 Is your strain/background tumor resistant? 

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• Unexpected/undesired  14

Mouse tumor phenotypes (in this discussion) Emphasize – Hematopoietic – Lung – Mammary – Liver

Hematopoietic Tumors Mouse V Human  Lymphomas

 Because

– Thymic L – Follicular B cell

– Top few spontaneous tumors • You should expect them

– Important human tumors too – Interesting models • Spontaneous + induced • GEM

 Lymphomas

 Retroviruses as

 FCC lymphomas

 Spontaneous tumors impact diverse studies  Good to know something about them …

T-cell lymphoma

B-cell lymphoma

Histiocytic sarcoma

Myeloid leukemia

Systemic (generalized)

Common

Rare

Rare

Rare

Thymus

Primarily

Rare

Rare

Rare

Spleen

Rare

Common–follicular or marginal zone

Occasionally – red pulp origin

Common

Peyer's patches

Rare

Common

Occasional

Rare

Mesenteric Lnodes

Rare

Common

Occasional

Rare

Liver

Rare

Rare

Common

Rare

Uterus

Rare

Rare

Common

Rare

Peritoneum

Rare

Rare

Common

Rare

Skin

Rare

Rare

Occasional

Rare

Bone marrow

Rare

Rare

Rare

Occasional

IHC

CD3

CD45r (B220) Pax5

CD68, F480

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 Activated Oncogenes  Inactivated Tumor Suppressors

Mouse Hematopoietic Tumors Spontaneous #1

Distribution

17From Table 1 - Ward 2006 Lymphomas and leukemias in mice. Experimental and Toxicologic Pathology,

– Diffuse large B cell

• Point mutations 16

Mouse Hematopoietic Tumors Spontaneous

– ALL in children – AML in adult

 Histiocytic sarcoma genetic engineers

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 Leukemias

 Thymic lymphoma (T lymphoplastic Lymphoma) – B cell tumors are rare in thymus

 Early onset often < 1y – Thymus 1st  disseminates

 AKR, C58, scid, NODscid –  Short studies …

 Kras activation  Endogenous retroviruses 18 Common (early) tumor after irradiation 3 of 15

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Mouse Hematopoietic Tumors Spontaneous #1  Clinical Signs:  Gross Lesions:

Mouse Hematopoietic Tumors Spontaneous #1 Histology

 Overall ‘blue – Starry sky ?

 Blue cells +  tingible body macrophages’ 19

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Mouse Hematopoietic Tumors Spontaneous #1 Histology

Historical Perspective: Furth 1946 - Ak strain (AKR)

Higher ‘Magnification’

 Overall ‘blue’ dt blue cells = mostly nucleus

 ‘Starry sky’ – Tingible body macrophages’ • Big macrophages with • Crumbs of nuclear debris 21

Historical Perspective: Hoag 1963

Mouse Hematopoietic Tumors Spontaneous #2 Follicular B cell lymphoma – Not exactly FCC -

 Most common tumor or lymphoma in some strains  GALT, nodes, spleen etc  Later onset often > 1y  Expect it in control mice > 1yr  K-Ras activation  Endogenous retroviruses 24

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Mouse Hematopoietic Tumors Spontaneous #2  Clinical Findings:  Gross Lesions:

Mouse Hematopoietic Tumors Spontaneous #2 Lymph node – Blue tumor – Monotonous population of similar cells with not much cytoplasm

Lymphoma. 25

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Mouse Hematopoietic Tumors Spontaneous #3

Yumoto & Dmochowski (1967) SJL/J reticulum cell neoplasms (lymphoma) C-type retrovirus particles

– Leukemia ?

Histiocytic Sarcoma  Most common tumor in some strains  Liver Uterus spleen nodes etc  Later onset often > 1y  Expect it in control mice > 1yr  Not such a blue tumor …  Not so round cells either …

28 Clker.com

Mouse Hematopoietic Tumors Spontaneous #3

Mouse Hematopoietic Tumors Spontaneous #3

Histiocytic Sarcoma – Organomegaly • Liver • Uterus • Invades anywhere

– Pleomorphic sarcomatous – Not so blue – Erythrophagocytosis 29

• Can be prominent

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 HS – not so blue 30

– Very nasty too

– Very Pleomorphic here

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Mouse Hematopoietic Tumors Spontaneous  Lymphomas & histiocytic sarcoma

Mouse Hematopoietic Tumors + Carcinogens  MORE FASTER

– Especially in old mice & susceptible strains

Accelerate/increase tumors in susceptible mice • Point mutations that activate oncogenes • Point mutations that suppress tumor suppressors

 Leukemias not so common except in advanced stage of lymphoma or histiocytic sarcoma with marrow involvement

Ulcerative dermatitis and nasty infections are much more common/likely causes of elevated WBC in mice. 31

 Hematopoietic tumors especially Thymic lymphoma are induced by chemical carcinogens and irradiation

 Increased tumors in strains that don’t get many ….

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Mouse Hematopoietic Tumors Genetically Engineered

– Carcinogens? Helicobacters? … – Genetic contamination by a susceptible mouse strain …. – Someone ordered the wrong strain …

Example of GEM Leukemia Tg.AC

Designer leukemias – Tg – insert human GENE of interest – Tm – knockout or disrupt mouse Gene of interest – Conditional mutants – express where you want, when you want – in theory …

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• • • • •

Tg. AC etc activated RAS or Ras Tp53 BCR-ABL Notch 1 Myc MYC

 Blue tumor  Erythroleukemia in marrow (G) lung (H)

 vs Lymphoma vs myeloid leukemia  IHC etc

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Hematopoietic Tumors Hu  Mouse Xenografts  Cell lines – Or Primary - from a patient – Do you MAP or PCR test for pathogens ?

LUNG

MARROW

 Activated Ras  (Skin tumor susceptible)  Bach & al. 2010 (ToxPath)

Hematopoietic Tumors Hu  Mouse Xenografts  Desired Outcome:  Splenomegaly, leukemia, death  Harvest tissues for FACS etc

 Orthotopic vs ectopic – Which site is best?

 Not Desired outcomes

 Which mouse is best?

 Early death  Prolonged Survival

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Imunodeficient mice made xenografts feasible…

Some Immunodeficient Options T Cells

B Cells

(nude)

D

Prkdcscid (scid)

D

Mutations Foxn1nu

PrkdcscidLystbg (scid-bg) NOD-Prkdcscid (NOD-scid)

 Challenges in immunodeficient mice

NOD, Prkdcscid, Il2rgtm… (NSG, NOG)

– Immunodeficient  Infections … – Leaky ?  B & T cells – NK etc cytotoxic cells – Tumors  mortality

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•

Leaky

N

N

yes

D

N-

yes

Thymic lymphoma

D

D

N

Reduced by bg

Thymic lymphoma

D

D

D

Minimal

Thymic lymphoma

D

D

D

No

N = Normal

D= Deficient

 Leakiness, NK cells & tumors can be problematic  Phenotypes vary with genetic background

 Post Xenograft Mortality   1 week

Innate NK

 Post Xenograft Survival   5 months after irradiation + IV Tail v Xenograft leukemia

post xeno

 Bactrim Diet x5m (trimethoprim sulfa)

Liver  Gram stain  Usually not so obvious  Bacteremia sepsis  Streptococcus? Enterococcus?

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 ‘NOG’ - CIEA Japan – truncated IL2rG (Ohbo et al. 1996),

 ‘NSG’ - Jax? Schultz ? – IL2rG null (Shultz et al. 2005). 40

 Post Xenograft Survival 

Xenograft ? Or mouse? origin  Spine

 Post irradiation + 6m Bactrim Rx.  Could this cause a clinical problem?

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Tumors

12BraytonCancerPhenoModel

Mouse Lung Tumors Spontaneous

Mouse Lung Tumors Terminology MMHCC: “Lung, adenoma or carcinoma” • Withhold the term ‘Bronchioloalveolar’

INHAND/Tox: A/B is still used • ‘Alveolar/Bronchiolar’ - Descriptive

Human BAC = adenocarcinoma with pure bronchioloalveolar growth pattern, no stromal, vascular or pleural invasion. – Lepidic pattern ~ aerogenous 43

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Mouse Lung Tumors Spontaneous

dissemination or lepidic spread

Historical Perspective: Hoag 1963

 Adenomas more common than Carcinoma

 Adenoma - smaller – Patterns: Solid > papillary > mixed

 Carcinomas - bigger – Papillary > mixed – May metastasize to liver  Nikitin, & al. (2004).

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 Wang & al. 2006.  Look like spontaneous but  Kras+/- &TP53 TG

Historical Perspective: Malkinson 1989

Lung Tumors Mouse V Human  Common in A >> 129, FVB/N >> B6, DBA

 Peripheral • Bronchiolar v alveolar v clara cells

 Most common neoplasm?  Most common CA death  NSCLC (80–85%) – Adca increasing – Smoking carcinogens

 Adenoma > Carcinoma  SCLC (15–20%)  Kras activation • Most common mutation in  KRAS activation (30-50%) adenoma & carcinoma  TP53 inactivation  Tp53 inactivation 48

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• Increased in carcinogen induced tumors

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Mouse V Human proliferative lesions of the mouse lung Nikitin 2004 – MMHCC consensus      

Human Lung Tumors Genes Activated

Hyperplasia is better described in the mouse.

Activated

Hyperplasia + adenoma are main proliferative lesions of mouse lung. Induced mouse tumors are usually multiple. Stromal response (fibrosis and inflammation) is weaker in mouse

KRAS in 20%-30% of BAC & ∼20% of all NSCLCs

Mouse tumors rarely metastasize.

(G-T transversions correlated with smoking)

Not in (non GEM) mice – – – –

Inactivated

Spontaneous (primary) squamous cell and neuroendocrine carcinomas Carcinomas with pleomorphic, sarcomatoid, or sarcomatous elements Carcinomas of salivary-gland type Combinations of tumor types, such as adenocarcinoma and Squamous or neuroendocrine carcinoma

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MYCN, MYCL or CMYC in SCLC, NSCLC

• Point mutations that activate oncogenes • Point mutations that suppress tumor suppressors

p16INK4A in CDK pathway

BCL-2 in SCLC> SCC, BAC

‘oncogenes ?’

Mouse Lung Tumors + Carcinogens Accelerate/increase tumors in susceptible mice

(GT transversions especially associated with smoking) RB protein found in >90% of SCLCs and 15%-30% of NSCLCs

Notch3

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 MORE FASTER

TP53 in 75% SCLCs > 50%NSCLC

‘Tumor suppressor genes?

Mouse Lung Tumors & Carcinogens Carcinogen

Human Exposure

Mutations

Human Associations

Cigarette smoke

Cigarette smoke

Kras, Tp53 etc Lung tumors etc

Lung + throat SCC ets

Cumene

Occ hazard

Kras MapK etc Tp53 etc

Lung tumors

Risk ? Risk ?

Mice

AZT

HIV Rx

Kras, Tp53 etc Lung tumors

• Cigarette smoke works well. • B6 & DBA are resistant. • Are these the best strains for your study ?

EtO

Sterilant

Kras

• Kras (& Tp53) mutations common in

1,3Butadiene- or Chloroprene

auto exhaust tobacco smoke

Kras, Tp53 etc Tumors, lymphoma,

induced tumors

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Mouse Lung Tumors & Carcinogens E.g. Cumene

hemangiosarcoma

Leukemia, lymphoma,

Designer lung tumors – Tg insert human GENE of interest – Tm knockout or disrupt mouse Gene of interest – Conditional mutants – express where you want,

 Mouse lung tumors  / inhibiting / downregulating

Kras & Mapk

Tp53

Invasion /metastasis Kr8, Kr18, Lasp1, Scc1, Mm14 Metastasis Sdc1, Ccnd1

Tumor suppressors Ptprd, Igs4a, Fh11, Pdzd2, Cdkn2d, Cdh5, Lox11, Akap2

Inhibit apoptosis Areg, Cks1b Increase angiogenesis Slc2a1, Gnb 211, 53 Ptges

Lung & Brain

Leukemia, lymphoma, Breast Ca, Stomach Ca

Mouse Lung Tumors Genetically Engineered

– assoc w hu lymphoma, leukemia

 / activating / upregulating

Lung, harderian, uterine tumors

Invasion inhibitors Reck, Gsn, Lims2,Cav1, Gpx3

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when you want - in theory … • Lung epithelial specific promoters + ____

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 Kras  Tp53 knockout or Insert mutant TP53  Rb or P16Inka inactivation

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Mouse Mammary Tumors Spontaneous

Lung Tumors Xenograft Hu  Mouse Cell lines e.g.

C3H !! Developed to study these

• Human SCLC Lu-24, Lu-130, H-69. • Human lung adenocarcinoma A549. • Human NSCLC H460 ( bone too)

– Or Primary – from a patient – Do you MAP or PCR test for pathogens ?

– CBA developed as resistant ‘control ‘strain

GR, BALB/c, 129 etc FVB/N get hyperplasia, EMT MMTV Bittner agent

Orthotopic vs ectopic – Which site is best? 55 Which mouse is best?

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Historical Perspective: Hoag 1963

– Reduced incidence by fostering on other dams – Transmitted in milk – cytoplasmic inclusions – Endogenous Mtv’s involved in later tumors too

Genetic Engineering by Retroviruses .. MMTV activates Notch1, Wnt & Fgf3   Characteristic morphologies • Described in 1911 by Haaland • Classified by Dunn 1958-9: A, B

Tg that activate WNT & FGF  morphologically similar tumors – Tumor types and acinar origin are not so Before fostering to eliminate exogenous MMTV

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relevant to Human breast Ca but we can activate more human relevant oncogenes or deactivate more relevant tumor suppressors

MMHC Annapolis Mammary classifications

MMTV Mtv1-55 = ‘genes’ in MGI Mtv proviruses vary in mouse strains Mtv1-3 were 1st associated with gene expression & tumorigenesis  Shed in milk etc secretions + inherited MMTV–like env found in 70% of human Breast cancers? 59 [email protected]

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Cardiff & al. 2000 Cardiff 2001 10 of 15

12BraytonCancerPhenoModel Annapolis (MMHCC) nomenclature 2001 Hyperplasia, Functional, without atypia Acinar Hyperplasia, Low Grade, Focal, Non-GEM

Nomenclature from Dunn 1958 & al Hyperplasia

Alveolar /Ductal

Hyperplastic alveolar nodule (HAN)

Alveolar

Solid Hyperplasia, Low Grade, Focal, Plaque/organoid/pregnancyHormone-induced, Non-GEM dependent "tumor" (Type P) Mammary Intra-epithelial Neoplasia No reports (GEM only) (MIN) Adenoma/Carcinoma* Adenocarcinoma Glandular/Acinar Type A Cribriform Type AB/Type B/Type L Papillary Type B/Type Y Solid Type B/Type P Adenoacanthoma/Pale cell Adenosquamous tumor/Large cell tumor EMT – epitheial mesenchymal Carcino sarcoma transitional tumor * 61

MMHCC Classification / Terminology

Cell origin / Differentiation

‘Preneoplastic’ lesions • HAN Hyperplastic alveolar nodules • Plaques • MIN mammary intraepithelial nodules – GEM

Ductal Alveolar /Ductal Alveolar Alveolar Alveolar Alveolar Alveolar/Ductal

Adenoma or carcinoma, – Pattern descriptors, – Etiology descriptors

Alveolar Alveolar

Modified/updated from Cardiff 2000, http://ccm.ucdavis.edu/bcancercd/contents.html

– E.g. Adenoma, acinar, MMTV (Wnt, Fgf3…) 62

Mouse Mammary Tumors Patterns  MMHCC – Acinar – Glandular – Cribriform – Papillary – Solid – Mixed – Adenosquamous

 Thelma Dunn 1958

Mouse Mammary Tumors Patterns Dunn 1958 Type A – (Acinar)

~ Polanti 1906 ~ Haaland 1911

Patterns A, B, C A acinar – common in spontaneous/MMTV B mixed – common in spontaneous/MMTV C cystic – older MMTV neg mice

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MMHCC: Acinar, glandular MMTV ‘spontaneous’ + Wnt Fgf3 activated GEM http://tgmouse.compmed.ucdavis.edu/preprint/ Kenney/Dunn1958.pdf

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Mouse Mammary Tumors Patterns Dunn 1958 Type B – mixed (Both)

MMHCC: acinar, glandular, papillary, cystic, solid MMTV ‘spontaneous’ + Wnt Fgf3 activated GEM 65

 ttp://tgmouse.compmed.ucdavis.edu/preprint/Kenney/Dunn1958.pdf

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Mouse Mammary Tumors Patterns Dunn 1958 Type C – Cystic

MMHCC: Glandular, Cystic

Not So common Old mice, MMTV neg 66 http://tgmouse.compmed.ucdavis.edu/pr

eprint/Kenney/Dunn1958.pdf

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Mouse Mammary Tumors Patterns

Mouse Mammary Tumors Patterns

Dunn 1958

Haaland 1911, Dunn 1958

Adenoacanthoma MMHCC: adenosquamous

MMHCC 2000:

Carcinosarcoma Carcinosarcoma

Cardiff etc 2010: EMT

Carcinogen associated – E.g. Methylcholanthrene BALB/c ? FVB/N ?  http://tgmouse.compmed.ucdavis.edu/p

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reprint/Kenney/Dunn1958.pdf

– 2013 MMHCC

http://tgmouse.compmed.ucdavis.edu/p 68

Mouse Mammary Patterns Carcinosarcoma now EMT

reprint/Kenney/Dunn1958.pdf

Mice have 10 Mammary Glands

 Spontaneous in FVB/N with pituitary prolactinomas – estrogen receptor alpha (ERalpha)-positive – FVB/N get mammary hyperplastic lesions that could interfere with Tg models

 Induced by hydrocarbons etc  Seen in relapsed or de-induced GEM tumors 69

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Mouse Mammary Tumors + Carcinogens

MOUSE Mammary Tumors Tumors  C3H, etc

Hyperplasia  Pregnant?  FVB/N – pituitary tt?

 MORE FASTER Accelerate/increase tumors in susceptible mice • Point mutations that activate oncogenes • Point mutations that suppress tumor suppressors

– B6 are generally resistant – Is it a great strain for your model?

MMTV

– Helicobacter hepaticus increased mammary

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tumors in mouse model -- Rao & al 2006

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Mouse MMTV-ERB-B2 or Human with ERB-B2 mutation ?

Mouse Mammary Tumors Genetically Engineered Designer MAMMARY tumors

MOUSE

when you want - in theory …

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• • • •

MMTV ltr is a GREAT mammary specific promoter Notch1, Wnt, Fgf3 etc

Cardiff

– Tg insert human GENE of interest – Tm knockout or disrupt mouse Gene of interest – Conditional mutants – express where you want,

HUMAN

HUMAN

MOUSE

Neu, Erb-B2 etc Tumor suppressor inactivation Brca1 & SV40Tag have poorly differentiated morphology-

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Mouse Mammary Tumors Xenograft Cell lines

Mammary Tumors Xenograft Hu  Mouse Foxn1nu/nu hypothymic mice lack T cells

• MCF7 etc require estrogens • MDA236, MDA468 etc

– Or Primary – from a patient – Do you MAP or PCR test for pathogens ?

– Not the only hairless mice…

Ectopic v Orthotopic

Orthotopic vs ectopic – Which site is best? Which mouse is best? 75

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Mammary Tumors Xenograft #1 NODscid MDA 468 xenograft

Ectopic subcutis  lung  77

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Mammary Tumors Xenograft #2 3mo F NODscid Estrogen pellet MCF 7 xenograft – Est dependent – Nice tumor 

Failure to thrive Ear/eye pallor 78Watery blood 13 of 15

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Mouse Liver Tumors Spontaneous

Bone marrow MDA 468 

MCF7 (+ estrogen) 

M >>F C3H, B6C3F1, CBA > >> other strains

Hcs loci – hepatocarcinogen

 Happy hematopoietic marrow

sensitivity

 Too much bone Not much marrow 79

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Mouse Liver Tumors Spontaneous Gross – nodules: single to multifocal coalescing Histology – Foci of cytoplasmic alteration • Eosinophilic + Clear cell > Basophilic or mixed

– Adenomas more common than CA – Carcinomas may arise in adenomas – Hepatoblastomas may arise in Ca 81

Mouse Liver Tumors + Carcinogens MORE FASTER Accelerate/increase tumors in susceptible mice • Point mutations that activate oncogenes • Point mutations that suppress tumor suppressors

Chemically induced models mimic injuryfibrosis-malignancy progression in humans.

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Mouse Liver Tumors + Carcinogens MORE FASTER Accelerate/increase tumors in susceptible mice • Point mutations that activate oncogenes • Point mutations that suppress tumor suppressors

Helicobacters – H hepaticus discovered when tumor resistant A/J mice developed liver tumors 83

 mutations in Hras, Catnb etc – Aflatoxin, CCL4, Choline deficient diet etc

Microbial agents (& Rx) & cancer models ? Helicobacters – liver gut (mammary) tumors, MALTomas ? Parvoviruses – target cells in S – Oncotropic, oncolytic – therapeutic ? ? Mycoplasma CARbacillus – lung lymphoma ? – Lymphocyte mitogens ? Bendazoles – Fenbendazole pinworm Rx – Microtubule inhibitors 84

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Mouse Liver Tumors Genetic Engineering Designer LIVER tumors

Mouse Liver Tumors Xenograft Cell lines e.g. – Hep3b (NB8064); SK-Hep (HTB-52 –

– Tg insert human GENE of interest – Tm knockout or disrupt mouse Gene of interest – Conditional mutants – express where you want,

endothelial features…); Hu-H7 – Or Primary – from a patient – Do you MAP or PCR test for pathogens ?

when you want - in theory …

• Ras, C-myc, Ctnb, Tgfa, Hcv etc  not so much injury/fibrosis 85

Orthotopic vs ectopic – Which site is best? Which mouse is best? 86

Conclusions Aims 1. Human & rodent neoplasms are not Identical –

Conclusions 4. Unexpected findings can be important!

BUT can be: • •

relevant to model mechanisms & treatments..

2. Know the options. 3. Know your model: • •

5. Pathology can help our Research.  Confirm & characterize expected model phenotypes & results.  Diagnose unexpected phenotypes or results.

Similarities to and differences from conditions being modeled. Know expected findings in your model  

•

 modify/improve experimental design

relevant to identify carcinogens..

Including genetic backgrounds & expected tumors Including the environment and factors that can complicate your project

Use Sufficient and relevant concurrent controls

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Is it ? Or isn’t it?

Conclusions Don’t shoot (or fire) the pathologist when its not the tumor you were aiming for …

GREAT TUMOR MODELS ?? $€ $€ Prix ??

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[email protected]

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