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
The rise of engineered T cells as cancer drugs
• Safety • Efficacy
• Potency • Specificity • Long-acting
• 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”.
Brentjens et al, Nat Med, 2003
Human CD19 CAR-targeted peripheral blood T cells eradicate systemic B lymphoma and ALL in mice
Br
B B
K
Br
H H V
C
Tumor Free Untreated 4 weeks
Day ≥ 5 Inoculation
WAVE Bioreactor
Selection ClinExVivo MPC
CAR+ expanded T cells
Day 3- 4 Transduction
SFG- CAR Vector
Apheresis Patient T cells
Incubation Dynabeads CD3/28
CD3+ enriched activated
T cells
DAY ≥ 1 0 Debeading
ClinExVivo MPC
Wash (Cell Saver5)
CAR transduced
T cells
Cryopreservation
Biosafety/ QC release tests
Patient Infusion
Validation In vivo antitumor activity
in SCID mice, CTL
CAR T cell Manufacturing Flow
Hollyman et al, J. Immunother, 2009
Przybylowski et al, Gene Ther, 2006
Day 0 Apheresis Thaw/Wash (Cell Saver5)
Isabelle Riviere Xiuyan Wang
Number of Patients N=45 (%) [95% CI]
Overall CR Rate
Morphologic disease (≥5% blasts) Minimal disease (<5% blasts)
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)
Relapsed, chemo-refractory adult ALL Summary of Clinical Outcomes (MSKCC, ASH 2015)
Overall Survival By MRD Status After CAR T Cell Treatment
Time Since CAR T Cell Infusion (Months)
Bone Marrow Transplant
Graft vs
Leukemia
Tumor Immunology
Adoptive Immunity
Natural ACTs • LAK • TIL • DLI • VST
Define and Isolate Natural T Cells
Gene Transfer
(Gene editing)
Chimeric antigen
receptors
(Synthetic biology)
Engineered ACT • The CD19 Paradigm
• Armored CARs
Design and Manufacture T Cells
Sadelain, Curr Opin Immunol, 2016
3rd gen 2nd gen – armored (CL)
1st gen 2nd gen
• 2nd gen CARs • CAR + X (armored CARs)
• CD4/CD8 ratio (defined composition) • T cell subset (eg TCM)
Sadelain and Mulligan, ICI, 1992
• Retroviral vectors (gRV, LV) • Transposons (Sleeping Beauty)
Meganuclease CRISPR-Cas9
Zinc-finger nuclease (ZFN) TALE nuclease (TALEN)
T-iPS-derived CAR-targeted T cells
Engineered T-iPS
Engineered antigen-specific T
lymphocytes
T-iPS
+
Chimeric Antigen
Receptor (CAR)
Hematopoietic specification and expansion
T lymphoid commitment
day 0 day 1
BMP-4
day 4
BMP4+bFGF
Mesoderm formation
VEGF, bFGF, SCF, FLT3L, IL-3
day 8 day 10
SCF, FLT3L, IL-3 OP9-DLL1 co-culture
SCF, Flt3L, IL-7
CAR+ T cell day 30
CAR+ T-iPS Day 22
TSP
ETP
β DJ
Pre-T
CD7 CD5
γδ VDJ γδ-T
TCRγδ CD4- CD8-/CD8αα
β VDJ Pre-Tα
ISP DP
Pre-TCR CD4 CD8α-/+
TCRαβ CD3 CD4 CD8αβ
α VJ
CD4SP
CD8 SP
TCRαβ CD3 CD4
TCRαβ CD3 CD8αβ
apoptosis
CD4SP
T helper
CD4SP T regulatory
CD8 SP
CD8 SP
T effector
T memory
MHC positive
selection
How T cells are made (in the thymus)
Themeli, Cell Stem Cells, 2015
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
Pre-
T ce
ll Da
y 7
Day
14
Day
18
Day
22
Day
35
Day
50
p=0.042
p=0.0038
p=0.0005
In vivo
CD19+ Raji-FFLuc i.p. Themeli, Nature Biotechnology, 2013
A perspective for large-scale generation of potent histocompatible therapeutic T cells
Themeli, Riviere & Sadelain, Cell Stem Cells, 2015
From idea to proof-of-concept to clinical benefit CD19 CAR therapy 1992-2013
Mouse T cell Engineering Sadelain and Mulligan, 1992
CD19 CAR therapy POC Brentjens et al, 2003
Clinical trials FDA Breakthrough design., 2014
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
Public education and network building
TNS9.3.55 collaborative network for the cure of beta-thalassemia
Seattle New York
MSKCC - NYPH NIH
Palermo Cagliari Rome Thessaloniki
Beirut
Farid Boulad New York
Isabelle Rivière New York
Aurelio Maggio Palermo
John F. Tisdale Bethesda
Vip Viprakasit Bangkok
Renzo Galanello Caglari
Bangkok
