RIP-Tag2 mouse model as a Paradigm for Target. Search in NETs

Transcription

1 RIP-Tag2 mouse model as a Paradigm for Target Search in NETs Oriol Casanovas, Ph.D. Tumor Angiogenesis Group INSTITUT CATALÀ d ONCOLOGIA IDIBELL Barcelona (SPAIN)

2 Therapeutic Targeting of the Tumor Stroma Joyce J, Cancer Cell 2005

3 Mechanisms of Tumor Angiogenesis

4 First Preclinical Models of NET NET Cell culture Injection into mice BON cells (CJ Thompson 1991) GOT1 cells (Kölby & Nisson 2000) Develop Tumors

5 First Preclinical Models of NET BON, GOT1 cells injected subcutaneously into Nude Mice: GOOD PRECLINICAL MODEL OF CARCINOID TUMORS! CAVEATS: - Only models of Carcinoids are available - Artificiality of cell culture growth - Subcutaneous growth of tumors is a limitation: - Different tumor growth kinetics - Lack of proper tumor stroma Limitations of their use for target discovery!

6 Another model of NET: Transgenic Mouse Model of Insulinoma RIP-Tag2 Transgene: Rat Insulin Promoter SV40 Large + Small T antigen prb p53 Insulinoma Tumors Acinar Tumor Hanahan D. Nature 1985

7 RIP-Tag2: Multi-Stage Development of NET Normal (onc+) Hyperplastic/ Dysplastic Angiogenic Tumor <5 wks 100% 5-77 wks ~50% 7-12 wks ~10% wks 2-4% Hanahan D. Nature 1985

8 RIP-Tag2: Distinct Neoplastic Lesions Distinct neoplastic lesions in RIP-Tag pancreas: Hanahan D. Nature 1985

9 RIP-Tag2: Quantitation of Tumorigenesis RIP1-Tag2 Pancreas Normal Pancreas 10wk 12wk Normal Islets Angiogenic Islets Tumors

10 Stage-Specific Experimental Therapeutic Trial Designs Prevention 5-8 wks of age Intervention wks of age Regression wks of age Hyperplastic/ Dysplastic Angiogenic Tumor Prevention trial: Can angiogenic switching be prevented? Intervention trial: Can tumor formation be prevented or slowed? Regression trial: Can tumor growth be stabilized or regressed d and lifespan be extended?

11 Antiangiogenic VEGFR inhibitors are effective in the RIP-Tag2 model DC101: Anti-VEGFR2 Ab specifically blocks activation of mvegfr2 Casanovas et al. Cancer Cell 2005

12 Anti-VEGFR2 Specific Blocking Antibody is fairy effective DC101: Anti-VEGFR2 Ab specifically blocks activation of mvegfr2. 35 Angiogenic Switch 180 Tumor formation 160 Tumor Growth Number of Angiogenic "red" Islets * Tumor Burden (mm3) * Tumor Burden (mm3) * 0 Ctrl Anti-R2 0 Ctrl Anti-R2 0 Ctrl Ctrl Anti-R2 t=0 10 days Casanovas et al. Cancer Cell 2005

13 Perivascular cells, PERICYTES, in tumor vasculature PDGFR β / CD31 PDGFR β / lectin

14 The concept of multi-targeted inhibition of tumor angiogenesis Bergers, et al. (2003). Benefits of targeting both pericytes and endothelial cells in tumor vasculature with kinase inhibitors. J.C.I., 111:

15 The concept of multi-targeted inhibition of tumor angiogenesis Bergers, et al. (2003). Benefits of targeting both pericytes and endothelial cells in tumor vasculature with kinase inhibitors. J.C.I., 111:

16 Antiangiogenic VEGFR small molecule inhibitors Sunitinib, Sorafenib, Pazopanib, PTK787 Favine et al. NRDD 2007

17 Vascular Regression Kinetics on Sunitinib Treatment Antiangiogenic effects in the RIP-Tag2 islet cell cancer tumor model: Control Sunitinib 4d Sunitinib 7d Sunitinib 14d. Sunitinib 28d. Pericyte EC association After Sunitinib Treatment: Sunitinib 7d Sunitinib 14d Sunitinib 28d McDonald Lab Virginia Yao UCSF

18 Anti-Angiogenic (Sunitinib) therapies for Cancer Sunitinib treated Transgenic multistage mouse model of cancer: RIP-Tag2 (D.Hanahan) Resistance? Untreated, 14.5 w Sunitinib, 18 w

19 Anti-Angiogenesis: a non-resistant therapeutic approach? Chemotherapy AntiAngiogenesis Target: Tumor Cell Genetic Instability High adaptation Resistance Target: Endothelial Cell Genetically Stable Less Adaptation No Resistance?

20 DC101 anti-vegfr2 Blocking Antibodies in RIP-Tag2 Tumors DC101: Anti-VEGFR2 Ab specifically blocks activation of mvegfr Tumor Burden (mm3) * RESISTANCE TO ANTI-ANGIOGENESIS Control Control Anti-VEGFR2 Anti-VEGFR2 t=0 10 days 4 weeks tt 12 wks of age Casanovas et al. Cancer Cell 2005

21 Test FGF family in the Tumor Regrowth Phase t=0 10 days 4 wks Stasis Phase Regrowth Phase Anti-VEGFR2 blocking antibody FGF TRAP Adenovirus-sFGFR2iiib 160 p= Tumor Burden (mm3) Ctrl t=0 Ctrl 10 d Anti-R2 10 d Anti-R2 4 wks Anti-R2 FGF trap 4 wks

22 Tumor Responses to Anti-Angiogenesis Blocking of VEGFR2 with DC101 mab in the RIP-Tag2 model: VEGF Anti-R2 No Angiogenesis Hypoxia Anti-R2 VEGF other Factors Reactivation of Angiogenesis RESPONSE PHASE with decreased angiogenesis (v. density & permeability) and inhibition of tumor growth (tumor stasis ). RESISTANCE PHASE with reestablishment of the vasculature and tumor re-growth, that implicates FGFs and other pro-angiogenic factors. Casanovas et al. Cancer Cell 2005

23 Resistance to Anti-Angiogenesis therapies for Cancer P.Carmeliet Insight Review, Nature Dec. 2005

24 Tunmor Adaptation to Anti-Angiogeneic therapies ANTIANGIOGENIC THERAPY (inh. VEGF/VEGFR2) Vascularized Tumor with sufficient nutrients and oxygen supply Stop neovascularization, Vessel Pruning Decreased MVD, blood perfusion, Hypoxia anti-tumor effect. Tumor Reaction : - Adaptation - Hypoxia tolerance - Microenvironment adaptation RESISTANCE!!!

25 Responses to anti-vegfr2: Increased Invasive Phenotype Transgenic multistage mouse model of cancer: RIP-Tag2 (D.Hanahan) Antiangiogenic therapy: DC101 anti-mvegfr2 Blocking Antibody (1 mg/mouse twice a week IP) Control (end-stage) Anti-VEGFR2 1 week Anti-VEGFR2 4 weeks H&E 150 µm 150 µm 150 µm Ac Ac Ac Anti-T antigen T 50 µm T 50 µm 50 µm T Pàez-Ribes et al. Cancer Cell 2009

26 Adaptive Responses to anti-vegfr2: Increased Dissemination & Mets Pàez-Ribes et al. Cancer Cell 2009

27 Enhanced Metastasis in Intermitent and Continuous Sunitinib treatment! Sunitinib 40 mg/kg daily t=0 5 wks Anti-CD31 Control Sunitinib 5 weeks Invasion Incidence (%) * ** Vehicle Sunitinib 10 Anti-T antigen Anti-T antigen 0 Incidence of metastasis (%) Non invasive tumors Liver metastasis * Microinvasive tumors Highly invasive tumors LN metastasis Control Sunitinib Liz Allen & Douglas Hanahan (UCSF)

28 Conclusions and Perspectives In several animal models: -PNET tg -GBM orthotopic -BrCa human orthotopic -RCC human orthotopic -PanCa human orthotopic In some patients? ONGOING at ICO-IDIBELL: Which patients suffer this effects?

29 Acknowledgements External Collaborators Marta Páez* Mar Martínez Lozano Gabriela Jiménez Lidia Moserle Francesc Viñals Mariona Graupera Gabriel Capellà Univ. California San Francisco (US) Douglas Hanahan Gabriele Bergers Karolinska Institutet (Sue) Kristian Pietras Ins. Richerca e Cura del Cancro (IT) Enrico Giraudo Clinical Collaborators Ramon Salazar Àlex Teulé Margarita García