CAR-modified T cell Therapy: How do we get there?

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CTL019 cell Lentiviral vector Anti-CD19 CAR construct CD19 Dead tumor cell Native TCR CAR-modified T cell Therapy: How do we get there? Shannon Maude MD PhD Center for Childhood Cancer Research Children s Hospital of Philadelphia ISCT 2014 Annual Meeting: Quality and Operations Track 12 Managing the CAR-T Before the Horse: A Cell Therapy Success Story

Disclosures No financial conflict of interest Advisory role in clinical trial development for Novartis CART19 licensed by Novartis Sponsor holds patent for technology

Redirecting the Specificity of T cells Gene transfer technology is used to stably express CARs on T cells, conferring novel antigen specificity 1,2 CTL019 cells can thus be directed against any tumor cell that expresses the CD19 surface antigen T cell CTL019 cell CTL019 therapy takes advantage of the cytotoxic potential of T cells thereby killing tumor cells in an antigendependent manner 1,3 Persistent CTL019 cells consist of both effector (cytotoxic) and central memory T cells 3 Native TCR CD19 Dead tumor cell Anti-CD19 CAR construct Tumor cell 1. Milone MC, et al. Mol Ther. 2009;17:1453-1464. 2. Hollyman D, et al. J Immunother. 2009;32:169-180. 3. Kalos M, et al. Sci Transl Med. 2011;3:95ra73.

(Not your typical) Enrollment process Manufacture Screening Leukapheresis Enrollment Stabilization Infusion CTL019

Patient Selection Population: Patients with multiply relapsed or highly refractory ALL Time from screening to treatment weeks to months Need to stabilize and maintain eligibility

Case #1 9 year old female with multiply relapsed ALL ALL History MUD PSCT for induction failure Relapsed post SCT Responded to reinduction Timing Month -2 Key events T cells collected (one week after completing reinduction chemotherapy) 6 Presentation Title Presenter Name Date Subject Business Use Only

Case #2 22 year old male with 1 st relapse of ALL ALL History 1 st relapse in maintenance therapy Refractory to reinduction chemotherapy and clofarabine/cytarabine Timing Month -2 Key notes T cells collected after failed reinduction 7 Presentation Title Presenter Name Date Subject Business Use Only

Timing of Leukapheresis Known eligible: Potentially eligible: Multiply relapsed/r efractory Relapsed after allo SCT Not eligible for allo SCT Refractory to multiple intensive therapies High-risk disease Primary induction failure Early BM relapse High-risk cytogenetics

Timing of Leukapheresis Known eligible: Potentially eligible: Multiply relapsed/r efractory Reserve for patients likely to need and benefit High-risk disease Better collection Better T cell growth Intensive therapy may limit collection and T cell expansion May be difficult to stabilize patient Usually minimal May delay therapy May not be used

Screening Failures Inadequate ALC for collection Poor collection Poor T cell expansion Inability to maintain eligibility Organ function Infection Rapidly progressive disease Proceed to SCT

Patient Selection Population: Patients with multiply relapsed or highly refractory ALL Time from screening to treatment weeks to months Need to stabilize and maintain eligibility Conditions likely to preclude enrollment and/or infusion Pre-existing organ dysfunction Uncontrolled infection Rapidly progressive disease

Patient Stabilization Time from screening to treatment weeks to months Goals: Prevent rapid progression Avoid organ toxicity and infectious complications NOT to induce remission or reduce disease burden Options: Maintenance chemotherapy (steroids, monthly or weekly VCR, +/- oral 6-MP and MTX) Hydroxyurea No chemotherapy if low white count and asymptomatic

Case #1 9 year old female with multiply relapsed ALL ALL History MUD PSCT for induction failure Relapsed post SCT Responded to reinduction Timing Month -2 Month -1 Key events T cells collected (one week after completing reinduction chemotherapy) No intervening chemotherapy 13 Presentation Title Presenter Name Date Subject Business Use Only

Case #2 22 year old male with 1 st relapse of ALL ALL History 1 st relapse in maintenance therapy Refractory to reinduction chemotherapy and clofarabine/cytarabine Timing Month -2 Month -1 Key notes T cells collected after failed reinduction Started hydroxyurea 14 Presentation Title Presenter Name Date Subject Business Use Only

Lymphodepleting chemotherapy Given 1 week prior to infusion Purpose: Induce lymphopenia to facilitate engraftment and homeostatic expansion of CTL019 T cells Disease control Agents: Cyclophosphamide 500 mg/m 2 IV daily Days 1-2, Fludarabine 30 mg/m 2 IV daily Days 1-4

Case #1 9 year old female with multiply relapsed ALL ALL History MUD PSCT for induction failure Relapsed post SCT Responded to reinduction Timing Month -2 Week -1 Day -1 Key events T cells collected (one week after completing reinduction chemotherapy) Received lymphodepleting chemotherapy (fludarabine/cyclophosphamide) Disease evaluation showed 5% blasts in bone marrow, no peripheral blasts 16 Presentation Title Presenter Name Date Subject Business Use Only

Case #2 22 year old male with 1 st relapse of ALL ALL History 1 st relapse in maintenance therapy Refractory to reinduction chemotherapy and clofarabine/cytarabine Timing Month -2 Month -1 Week -1 Day -1 Key notes T cells collected after failed reinduction Started hydroxyurea Received lymphodepleting chemotherapy (fludarabine/cyclophosphamide) Disease evaluation showed 97% blasts in BM, no peripheral blasts 17 Presentation Title Presenter Name Date Subject Business Use Only

CTL019 Infusion Premedication: Tylenol and Benadryl Infusion: Cell product thawed per Stem Cell Lab SOPs Outpatient infusion center Infused over 2 minutes by trained staff Vital signs monitored every 15 minutes for 1 hour Acute infusional toxicities rare

Cytokine Release Syndrome CRS is related to T cell expansion and is likely necessary for efficacy Symptoms typically occur 1-14 days after CTL019 cell infusion in ALL Fever Myalgias Nausea/Vomiting Hypotension Respiratory insufficiency Renal insufficiency Coagulopathy Severity scales with disease burden

CRS management Symptom management Antipyretics Analgesics (often narcotics required) Antiemetics, TPN Blood products (FFP, cryo) O2, CPAP, ventilation Vasopressors

Case #1 9 year old female with multiply relapsed ALL ALL History MUD PSCT for induction failure Relapsed post SCT Responded to reinduction Timing Month -2 Week -1 Day -1 Key events T cells collected (one week after completing reinduction chemotherapy) Received lymphodepleting chemotherapy (fludarabine/cyclophosphamide) Disease evaluation showed 5% blasts in bone marrow, no peripheral blasts 21 Presentation Title Presenter Name Date Subject Business Use Only

Temperature, C Case #1 CRS Timeline 42 CTL019 Infusion Temperature D/C home 41 40 39 38 37 0 1 2 3 4 5 6 7 8 Days (post-infusion) Narcotics (days 3-6) Myalgias (days 2-7) Antibiotics (days 1-7) 22 Presentation Title Presenter Name Date Subject Business Use Only

Severe CRS management Lympholytics Steroids tried with some effect but potential to reduce efficacy Cytokine-directed therapy IL-6 noted to be very elevated Anti-IL-6 therapy highly effective with no apparent effect on efficacy Grupp et al. NEJM 2103

Macrophage Activation Syndrome (MAS) /Hemophagocytic Lymphohistiocytosis (HLH) Systemic inflammatory response with vascular leak, hypotension, respiratory and renal insufficiency Extraordinarily high ferritin levels suggest MAS/HLH 16,000 to 415,000 ng/ml Coagulopathy Elevated D-dimer and low fibrinogen HSM, Transaminitis Genetic w/u patients 100 and 125, both gr 4 CRS, have a potentially predisposing perforin mutation Entirely reversible with cytokine blockade IL-6R blocking agent tocilizumab Abbreviations: MAS/HLH, macrophage activating syndrome/hemophagocytic lymphohistiocytosis

Tocilizumab IL-6 receptor antagonist Blocks IL-6 mediated effects Indicated in: juvenile idiopathic arthritis (JIA) Rheumatoid arthritis (RA) In Japan, indication for Castleman s Disease Typically given monthly Rare side effects of transaminitis and neutropenia Not required long-term, but a few patients have required 2 doses

Case #2 22 year old male with 1 st relapse of ALL ALL History 1 st relapse in maintenance therapy Refractory to reinduction chemotherapy and clofarabine/cytarabine Timing Month -2 Month -1 Week -1 Day -1 Key notes T cells collected after failed reinduction Started hydroxyurea Received lymphodepleting chemotherapy (fludarabine/cyclophosphamide) Disease evaluation showed 97% blasts in BM, no peripheral blasts 26 Presentation Title Presenter Name Date Subject Business Use Only

Temperature, C LDH, IU/Liter Case #2 CRS Timeline CTL019 infusion Day 0: 10% of dose Day 1: 30% of dose 42 41 40 39 38 37 36 Tocilizumab (days 5 and 8) Temp 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 Days (post-infusion) LDH 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Confusion (day 2-11) 27 Rasburicase (Day 9) Presentation Title Presenter Name Date Subject Business Use Only High dose steroids (days 7-11) Respiratory support (days 3-10) High-dose vasopressors (days 2-9) Transfusion support (days 2-15) Cryoprecipitate (days 10-15) FFP (days 2 and 8)

PT/ PTT, seconds Fibrinogen, mg/dl Case #2 - Coagulopathy CTL019 infusion Day 0: 10% of dose Day 1: 30% of dose 70 60 50 FFP (days 2 and 8) PT PTT Fibrinogen 700 600 500 40 30 20 10 0 Fibrinogen PTT PT 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 Days (post-infusion) 400 300 200 100 0 FFP, Platelets (days 2-15) Cryoprecipitate (days 10-15) 28 Presentation Title Presenter Name Date Subject Business Use Only

Neurotoxicity issues Seen in several CD19 trials: NCI CHOP/UPenn MSKCC Blinatumomab In CHOP/UPenn experience with CTL019: Self-limited, generally untreated, fully resolves??? Related to CRS Not prevented by tocilizumab

High Rates of Remission Induction in ALL Over 80 patients have been treated with CTL019 (CLL, ALL, and B-cell Lymphomas) Pediatric ALL cohort (N=25): 22/25 CRs (88%) 6 relapses, including 2 CD19(-) relapses All ALL (N=30) (pediatric plus adult): 27/30 CRs (90%) 7 relapses Short followup (median 6 months, range 2-21 months)

Probability Probability Event-free and Overall Survival Event free Survival Overall Survival 1.0 1.0 0.8 0.8 6 month OS: 78% (95% CI: 64,95) 0.6 6 month EFS: 66% (95% CI: 49,87) 0.6 0.4 0.4 0.2 0.2 0.0 0 90 180 270 360 450 540 630 N: 30 18 13 5 1 1 1 1 Days since infusion 0.0 0 90 180 270 360 450 540 630 N: 30 24 18 10 4 2 1 1 Days since infusion

Disease Burden at Time of Infusion Patient population 2 nd relapse Majority refractory to multiple prior therapies Abbreviations: BM, bone marrow; CR, complete response, MRD, minimal residual disease; NR, no response

Most patients treated POST allo 18 patients post-allo SCT T cells collected from patient No evidence of GVHD 6 months post-sct Median donor chimerism 100% No GVHD to date Abbreviations: GVHD, graft-versus-host, disease; SCT, stem cell transplant

Long-term Management B cell aplasia: Monthly IVIG Follow-up: 1 st year Close follow-up for CTL019 persistence and relapse Discuss SCT (particularly for patients in CR2) Long-term 15-year follow-up for gene therapy

ACC TRP Carl June Anne Chew Michael Milone Yangbing Zhao John Scholler Elizabeth Veloso Dana Hammill Grupp Lab David Barrett David Teachey Alix Seif Shannon Maude Sarah Tasian Junior Hall Jessica Hulitt Terry Ryan Yueh Chang Yong Li Jessica Lee It really does take a village CVPF Bruce Levine Julio Cotte Zoe Zheng Alexey Bersenev U Penn Clinical David Porter Noelle Frey CHOP Clinical/Study Staff Richard Aplenc Sue Rheingold Shannon Maude Christine Strait Colleen Callahan Manuel Litchman Angela Shen Patricia Wood Michele Sharr-McMahon TCSL Simon Lacey Michael Kalos Yolanda Mahnke Jos Melenhorst Minnal Gupta Irina Kulikovskaya Jeff Finklestein Frazana Nazimuddin Vanessa Gonzalez Patients and Families CHOP Nursing CTO/IND Office CHOP Stem Cell Lab Giuliana Pierson Yongping Wang CHOP Vector Core Frasier Wright Adaptive TcR