The promise of T cell engineering CD19 CAR therapy A prologue to immune regenerative medicine Vast potential, patience, public education

Transcription

1 The promise of T cell engineering CD19 CAR therapy A prologue to immune regenerative medicine Vast potential, patience, public education Academies Cell Therapy Workshop Washington, October 13, 2016 Michel Sadelain, MD, PhD Director, Center for Cell Engineering Immunology Program, Sloan Kettering Institute Departments of Medicine and Pediatrics Memorial Sloan Kettering Cancer Center New York, NY

2 The rise of engineered T cells as cancer drugs A major limitation of most existing cancer therapeutics is the lack of specificity or curative potential. Engineered T cells offer the prospect of combining specificity, potency and persistence from an optimized living drug. Safety Efficacy Potency Specificity Long-acting

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4 Human CD19 CAR-targeted peripheral blood T cells eradicate systemic B lymphoma and ALL in mice Br Br C K H V H B Tumor Free B Untreated 4 weeks Brentjens et al, Nat Med, 2003

5 CAR T cell Manufacturing Flow Incubation Dynabeads CD3/28 Day 3-4 Transduction SFG- CAR Vector CD3+ enriched activated T cells Przybylowski et al, Gene Ther, 2006 Day 0 Apheresis Thaw/Wash (Cell Saver5) Cryopreservation Apheresis Patient T cells Selection ClinExVivo MPC CAR transduced T cells Day 5 Inoculation WAVE Bioreactor Biosafety/ QC release tests Patient Infusion Validation In vivo antitumor activity in SCID mice, CTL Wash (Cell Saver5) DAY 1 0 Debeading ClinExVivo MPC Hollyman et al, J. Immunother, 2009 CAR+ expanded T cells Isabelle Riviere Xiuyan Wang

6 Relapsed, chemo-refractory adult ALL Summary of Clinical Outcomes (MSKCC, ASH 2015) Overall CR Rate Morphologic disease ( 5% blasts) Minimal disease (<5% blasts) Number of Patients N=45 (%) [95% CI] 37/45 (82%) [68 92] 18/24 (75%) [53 90] 19/21 (91%) [70 99] Overall MRD Negative CR Rate * 30/36 (83%) Mean Time to CR (SD) 22 days (9.4) * Assessed among those patients who achieved CR and evaluable for MRD analysis (n=36)

7 Overall Survival By MRD Status After CAR T Cell Treatment Time Since CAR T Cell Infusion (Months)

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9 Define and Isolate Natural T Cells Design and Manufacture T Cells Bone Marrow Transplant Graft vs Leukemia Tumor Immunology Adoptive Immunity Gene Transfer (Gene editing) Chimeric antigen receptors (Synthetic biology) Natural ACTs LAK TIL DLI VST Engineered ACT The CD19 Paradigm Armored CARs Sadelain, Curr Opin Immunol, 2016

10 1 st gen 2 nd gen 3 rd gen 2 nd gen armored (CL) 2 nd gen CARs CAR + X (armored CARs) CD4/CD8 ratio (defined composition) T cell subset (eg TCM) Zinc-finger nuclease (ZFN) TALE nuclease (TALEN) Meganuclease CRISPR-Cas9 Sadelain and Mulligan, ICI, 1992 Retroviral vectors (grv, LV) Transposons (Sleeping Beauty)

11 T-iPS-derived CAR-targeted T cells + T-iPS Chimeric Antigen Receptor (CAR) Engineered T-iPS Engineered antigen-specific T lymphocytes CAR + T-iPS Mesoderm formation BMP-4 BMP4+bFGF day 0 day 1 day 4 Hematopoietic specification and expansion VEGF, bfgf, SCF, FLT3L, IL-3 SCF, FLT3L, IL-3 day 8 day 10 T lymphoid commitment OP9-DLL1 co-culture SCF, Flt3L, IL-7 Day 22 day 30 CAR + T cell

12 How T cells are made (in the thymus) TSP β DJ ETP Pre-T CD7 CD5 γδ VDJ β VDJ Pre-Tα γδ-t TCRγδ CD4- CD8-/CD8αα ISP Pre-TCR CD4 CD8α-/+ α VJ DP TCRαβ CD3 CD4 CD8αβ TCRαβ CD3 CD4 CD4 SP MHC positive selection CD8 SP CD4 SP CD4 SP TCRαβ CD3 CD8αβ T helper T regulatory CD8 SP CD8 SP T effector T memory apoptosis Themeli, Cell Stem Cells, 2015

13 1928z-T-iPS-T cells lyse CD19+ tumor cells in vitro and in vivo In vitro No treatment 1928z-αβ 1928z-γδ 1928z+ T-iPS-T Day 50 Day 14 Day 35 Day 7 Pre-T cell In vivo p= Day 18 p= Day 22 p=0.042 Themeli, Nature Biotechnology, 2013 CD19+ Raji-FFLuc i.p.

14 A perspective for large-scale generation of potent histocompatible therapeutic T cells Themeli, Riviere & Sadelain, Cell Stem Cells, 2015

15 From idea to proof-of-concept to clinical benefit CD19 CAR therapy Mouse T cell Engineering Sadelain and Mulligan, 1992 CD19 CAR therapy POC Brentjens et al, 2003 Clinical trials FDA Breakthrough design., 2014

16 Cell Therapy and Cell Engineering Facility Isabelle Rivière, PhD, Director Non-profit; Grants, philanthropy, foundations Current GMP Facility: 6 cleanrooms New GMP facility opening 2014: 13 cleanrooms Cellular products (CAR T cells, HSCs, DCs, NK cells, cord blood, hes/da neurons, T-iPS), RNA vectors, plasmid DNA, CRISPR/Cas9 Phase I/II clinical trials Multicenter trials with other academic centers

17 Public education and network building TNS collaborative network for the cure of beta-thalassemia Seattle New York MSKCC - NYPH NIH Bangkok Rome Thessaloniki Cagliari Palermo Beirut Farid Boulad New York Isabelle Rivière Aurelio Maggio John F. Tisdale Renzo Galanello Vip Viprakasit New York Palermo Bethesda Caglari Bangkok