Model Mouse Of Human Cancer

Mouse models of human cancer

Marco Giovannini, MD-PhD

INSERM U674 «Génomique Fonctionnelle des Tumeurs Solides» CEPH - IFR105-Saintt Louis - Université Paris 7 IUH

 

 

Mouse models of human cancer Cancer in the mouse should look and act like the human disease Same gene and/or pathways should be affected in tumor initiation and progression Same or similar histological features of human tumors and they should progress through the same stages The response of a given tumor to a particular therapy in the mouse should accurately reflect the response in human patients van Dyke & Jacks, Cell, 2002

 

 

Tumor predisposition

 

Gene

human +/-

mouse +/-

RB

retinoblastoma

pituitary adenoma

APC

familial adenomatous polyposis

intestinal polyps

p53

Li-Fraumeni syndrome

Sarcoma, lymphoma others

NF1

neurofibromatosis type 1 (neurofibroma)

Myeloid leukemia, Pheochromocytoma

NF2

neurofibromatosis type 2 (schwannoma, meningioma)

Sarcoma (osteo-, fibro-)

WT1

Wilms’ tumor

None

Brca 1

familial breast /ovarian cancer

None

Brca 2

familial breast cancer

None

 

 

Gene

murine homozygous mutant phenotype

Rb

Embryonic lethality (E13.5-15.5)

Apc

Embryonic lethality (E5-6)

p53

10% embryonic lethality, viable mice develop lymphoma, sarcoma and other tumors

Nf1

Embryonic lethality (E13.5)

Nf2

Embryonic lethality (E6.5-7.5)

wt1

Embryonic lethality (E13.5)

Brca 1

Embryonic lethality (E5-6-E10-13)

 

model of human Schwann cell tumorigenesis

NF2 loss

schwannoma WHO grade I

Schwann cells neurofibroma

MPNST

WHO grade I

NF1 loss

 

WHO grade III-IV

P53 mutation  

Understanding NF2: insights from mouse models

 

 

Neurofibromatosis type 2

NF2-/-

meningioma

NF2 +/bilateral vestibular schwannoma

NF2-/-

ependymoma

 

 

The NF2 gene product: merlin/schwannomin Sequence divergent in isoform 1&2 304

100%

NH3+

COO- 595aa

a-helical domain

FERM-domain

prolin rich seq.

Ezrin

NH3+

61%

COO- 586 aa

Radixin

NH3+

63%

COO- 583 aa

Moesin

NH3+

63%

COO- 577 aa

N-terminal domain:

C-terminal domain :

Interaction with membrane  

Interaction with cytoskeleton  

Growth factor receptors

Stress fibers

Membrane  ruffling

Rho 

F­actin

Ras exchange Factors (SOS)

GAPs: p120RasGAP Neurofibromin (NF1)

Raf Rac

F­actin

PI3 kinase Akt

Sch/merlin (NF2) P P

 

Ras

McClatchey and Giovannini Genes & Development 2005

PAK

 

Nucleus

MEK ERK MAP kinase

1. Nf2+/- mice don’t show manifestations of human NF2

Nf2 +/- mouse

NF2 patient  

 

Giovannini et al.  Genes & Development 2000

Cre/loxP system

loxP

ATAACTTCGTATAATGTATGCTATACGAAGTTAT TATTGAAGCATATTACATACGATATGCTTCAATA loxP

loxP

Cre

 

loxP  

loxP

Neural crest cell

p75+++ P0+

Minimal rat P0 promoter (1,1kb) axons

p75+++ P0+

Schwann cell precursor Committed Schwann cell

p75+++ P0+

axon

p75++ P0+

Premyelinating Schwann cell Promyelinating Schwann cell

p75++ Krox20+ P0+

Myelinating Schwann cell

 

 

myelin p75+ Krox20+ P0+++

Nf2-related Schwann cell tumorigenesis in the mouse

90% Schwann cell hyperplasia

30%

schwannoma

P0-Cre;Nf2flox/flox

 

 

Giovannini et al., Genes & Dev. 2000

[CANCER RESEARCH 64, 3718–3724, May 15, 2004] Meeting Report Comparative Pathology of Nerve Sheath Tumors in Mouse  Models and Humans A O. Stemmer­Rachamimov, D N. Louis, G P. Nielsen, C R. Antonescu,  A D. Borowsky, R T. Bronson, D K. Burns, P. Cervera, M E.  McLaughlin, G. Reifenberger, M C. Schmale, M. MacCollin, R. C. Chao,  K. Cichowski, M. Kalamarides, S. M. Messerli, A. I. McClatchey, M.  Niwa­Kawakita, N. Ratner, K. M. Reilly, Y. Zhu,  and  M. Giovannini.

 

 

Preparing clinical trials NF2 Natural History Consortium

Coordinator: House Ear Institute, Los Angeles, CA Financed by a Dept. Of Defense, U.S. Army grant Responsable de l'étude française: Michel Kalamarides Attaché de Recherche Clinique: Marco Giovannini

Participants: Mass. Gen. Hosp., Mount Sinai NYU, U. Texas Houston, Ohio State U., Melbourne, Hamburg, Manchester, Paris (Hôpital Beaujon, AP-HP) Goals: • To determine the growth rate and standardize volumetric analysis of NF2 tumors • To associate the tumor growth rate with the genotype of NF2 patients • To standardize clinical data collection by providing an infrastructure mechanism for potential clinical trials

 

 

Laboratory of Functional Genomic of Solid Tumors Inserm U674 Marie-Claude Jaurand, Ph.D., DR1 Inserm Jessica Zucman, M.D., Ph.D., DR2 Inserm Dominique Lallemand, PhD, CR1 Inserm Michel Kalamarides, M.D, Ph.D., PU-PH, AP-HP Marco Giovannini, M.D., Ph.D., DR2 Inserm

Fondation Jean Dausset - CEPH IFR 105 Saint-Louis Université Paris 7 - IUH

 

 

Plateau Technique d'Expérimentation Animale Responsable: Michiko Niwa-Kawakita, PhD, IR (CEPH-Inserm*) Zhi-Yan Han, PhD, IR (Inserm*.) Fabrice Chareyre, AI (Inserm*) Martine Mauchauffé, IE1 (Univ. Paris VII)

Key interactions: Dr. Marika Pla (Dept. Exp. Animale, IUH/INSERM) Pr. Anne Janin (Service d’Anatomopathologie/INSERM) Pr. Hugues de Thé (Dept. Exp. Therap., ARECA, CNRS)

 

 

Drug sensitivity

Study of genetic and chromosomal instability

Lignées Mouse models cellulaires of human tumorales cancer

Development of new therapeutic vectors  

 

Study of tumorigenic pathways

Mouse Models of Human Cancer: from stem cells to therapeutic intervention Principal Investigator: Marco Giovannini, 26 teams (IUH, I. Curie, IGR, I. Pasteur, I. Cochin) Projet is part of axe 2 du « Cancéropôle I.d.F » Coordinator: H. de Thé

12 13

 

mouse mutant strains produced by consortium teams

 

Objective .1 Generation of new mouse models of cancer •

For cognitive studies

•

As preclinical tools

 

 

Technological Innovation A new protocol for the selection, cloning, and identification of targeted ES cell clones.

loxP

pgk prom

Hyg

EGFP

polyA

loxP

Advantages: •Sorting of ES cells in 96-well plates in a few minutes •M. Niwa-Kawakita, F. Chareyre, J Bayer  

 

Objective .2

Cancerepositoire : Collection of mouse models of general interest in cancer research

 

 

Objective.3 Génotyping « Congenissimo »

Speed Congenics : marker-assisted technique of selection allowing faster production of congenic mouse strains

 

 

Objective .4 Establishment of in vitro ex vivo cell systems

 Mouse tumors  tissues of GEM mice

 

 

Objective .5 Development of imaging protocols for monitoring tumor growth in preclinical trials

 

 

• • • • • • • • • • • • • • • • •

Michel Kalamarides*, MD, PhD Dominique Lallemand, PhD Marie Claude Jaurand, PhD Michiko Niwa-Kawakita, PhD Cosimo Martinelli, PhD Zhi Yan Han, PhD Odette Mariani, PhD Pascale Cervera, MD Joceline Fleury Feith, MD Françoise Barthes Le Pimpec, MD Jan Manent Stephane Goutagny Martine Mauchauffé Fabrice Chareyre Aurélie Lampin Aurélie Wattilieux Annie Renier

INSERM Unit674 «Functional Genomics of Solid Tumors » Human Polymorphism Study Center -CEPH- Paris7 IUH Hôpital Saint-Louis, *Hôpital Beaujon, Paris, France

Collaborations •

Consortium "preclinical mouse models of neurofibromatoses": A. McClatchey, T. Jacks, L. Parada, K. Shannon

•

NexGenix Pharma, Burlingame, CA

•

A. Berns, NKI, Amsterdam, NL

•

Rona Carroll, Peter Black Brain tumor lab, Brigham & Women's Hospital, Harvard, Boston, USA

•

David Gutmann Department of Neurology, Washington University, St Louis, MO

U.S. Army grants DAMD17-02-1-0645, DAMD17-02-1-0638? DAMD17-02-1-0653 James S. McDonnell Foundation French Neurofibromatosis Foundation Ligue Nationale contre le Cancer Association de Recherche contre le Cancer Canceropôle Ile de France Inserm