NW Symposium in Melanoma May 22, 2010 Adoptive T Cell Therapy

1. NW Symposium in MelanomaMay 22, 2010Adoptive T Cell Therapy Cassian Yee MD Member Program in Immunology Fred Hutchinson Cancer Research Center cyee@fhcrc.org

2. Melanoma Cell T Cell

4. Melanoma Cell T Cell

5. Tumor Cell T Cell TCR Target Antigen

7. WBCs WBCs RBCs platelets T cells B cells

8. Tumor Cell T Cell CD8+ TCR Target Antigen 1 in 100,000

9. CD4 CD8 DC DC DC DC Tumor Immune Surveillance Tumor Normal

10. Possible Reasons For Failure of Tumor Immune Surveillance • Immune evasion • T cell frequency : Tumor burden Highly functional T cells More of them

11. Vaccine Therapy Adoptive Therapy

12. CD4 CD8 DC DC DC DC Adoptive T Cell Therapy Tumor

13. LEUKAPHERESIS WBCs

17. Study in Patients with Advanced Melanoma Treated with T Cell Therapy • Recruited patients with • metastatic disease • progressive • failing 2nd and 3rd attempts to treat cancer

18. Before After T cell infusion Patient 1017-6 Patient 1017-8

19. Study in Patients with Advanced Melanoma Treated with T Cell Therapy • 30% Failed • 30% Partial Response • 30% Stabilized Response • 10% near Complete Response 3 to > 30 months

20. other growth signals other cytokines IL-15 IL-7 Lymphoid Homeostasis

21. Lymphodepletionbuilding a better environment Increase 'space' for transferred T cells Eliminate 'suppressor cells' Supply Growth Factors

22. PROTOCOL # 2140 Adoptive T Cell Therapy following Cyclophosphamide Lymphodepletion Objectives :Eligibility Criteria: - Evaluate Safety - Stage IV (Metastatic) - Evaluate T Cell Persistence - HLA-A2 - Evaluate anti-tumor efficacy T Cell Infusion: - Antigen-specific CD8+ T cell clones - Targeting MART-1, gp100 - Dose: 1010 cells / m2 CY 60 mg/kg x 2 Low-Dose IL-2 (250,000 U s.c q12 h)

24. T cell persistence in vivo 2140-1 5.3% 1.1% D18 2140-2

27. Clinical Response

28. Future Directions

29. Future of Adoptive T Cell Therapy • LATE-STAGE DISEASE • EARLY-STAGE DISEASE • COMBINATION THERAPY • WHICH CANCER TYPES?

32. Adoptive Therapy following Lymphodepletion TBI FLU 25 mg/m2 x 5 CY 60 mg/kg x 2 HD IL-2 720K u/kg TID High-Dose IL-2 (600,000 u./kgq8) Low-Dose IL-2 (250,000 U s.c q12 h)

33. Adoptive Therapy following Cytoxan Lymphodepletion • Protocol 2140 CY 60 mg/kg x 2 HD IL-2 720K u/kg TID High-Dose IL-2 (600,000 u./kgq8) Low-Dose IL-2 (250,000 U s.c q12 h)

34. Cytokine modulation • Phenotype • - CD8/CD4 • - Memory phenotype • TCR • Chimeric receptor • Costimulatory/Inhbitory modification • Suicide gene • Lymphodepletion • - Chemotherapy/TBI • Cytokine help • - Low-dose IL-2 • - High-dose IL-2 • - Other γ-chain receptor cytokines • Vaccine + adoptive therapy Translational Strategies to Augment the CD8 Effector Response Isolate/Enrich Clone/Select Genetically Modify Ê CD4 Intrinsic Tet-PE CD8-FITC Expand Post-infusion Immunomodulation Pre-infusion Immunomodulation Extrinsic Fludarabine Hematol Oncol Clin North Am. 2006 Jun;20(3):711-3

35. Chimeric TCR + zeta Chimeric Ig + zeta Receptor Transfer T Cell Expansion & Infusion • Adoptive Therapy Using Antigen-Specific T cells • Transferred Receptor Tumor Cell