Advances in Adoptive Cellular Therapy of Cancer. Melanoma Bridge Meeting December 5, 2014
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1 Advances in Adoptive Cellular Therapy of Cancer Melanoma Bridge Meeting December 5, 2014 David Stroncek, MD Chief, Cell Processing Section, DTM, CC, NIH Bethesda, Maryland, USA
2 Disclosures None
3
4 Focus of Recent Work has Been to Improve Several Areas of Adoptive Cellular Therapies Potency Safety Manufacturing Types of patients treated
5 Potency Critical in vivo biological function Measure of the Mechanism of Action of Cellular and Gene Therapies
6 Why is Potency Important? Clinical: Improve the clinical effectiveness of cellular therapies Manufacturing: Critical for product quality control Regulatory: FDA requirement
7 Factors Effecting TIL Potency
8 Results of TIL Therapy of Metastatic Melanoma: How Do We Improve them? SA Rosenberg, ME Dudley Curr Opin Immunol. 2009;21:233-40
9 Young TIL Telomere Length, CD27 Expression and Persistence Affect Patient Outcome Mean telomere length, the number of CD27+CD8+ cells, and the percentage persistence of the infused cells in peripheral blood at 1 month after cell infusion are significantly different in objective responders (CR + PR) compared with nonresponders (all P2 < 0.001) Rosenberg S A et al. Clin Cancer Res 2011;17:
10 TIL and Nonmyeloablative Lymphocyte Depleting Chemotherapy for Metastatic Melanoma: Overall Survival (Sheba Medical Center) 80 patients enrolled and 57 were treated Besser M J et al. Clin Cancer Res 2013;19:
11 Shorter Duration of Expansion and Infusion of more CD8+ Cells is Associated with Better Clinical Outcomes (Sheba Medical Center) Besser M J et al. Clin Cancer Res 2013;19:
12 TIL Therapy at MD Anderson: Change in Tumor Size in 31 Patients 15 responders; 4 complete responders Radvanyi L G et al. Clin Cancer Res 2012;18: by American Association for Cancer Research
13 TIL From Responders Contain More CD8+ T Cells Total TIL Infused % CD8+ Cells in TIL Total CD8+ Cells Infused Radvanyi L G et al. Clin Cancer Res 2012;18: by American Association for Cancer Research
14 TIL From Responders Contain More CD8+ T Cells Total CD8+ Cells Infused vs Change in Tumor Size Quantities of CD4+ Cells Did Not Affect Clinical Outcome Radvanyi L G et al. Clin Cancer Res 2012;18: by American Association for Cancer Research
15 Summary Better clinical outcomes of TIL therapy are associated with Greater percent of CD8+ T cell Greater numbers of CD8+ T cells Shorter time in culture
16 Manufacturing More Potent TIL
17 Manufacturing of Tumor Infiltrating Lymphocytes (TIL) Tumor Tumor Digest + IL-2 Patient with Metastatic Melanoma Transfuse 30 to 60 x 10 9 cells and administer IL 2 Expanded TIL Bulk TIL Isolation REP: IL-2 + Anti-CD3 + Feeder Cells
18 Addition of 4-1BB antibody in the TIL REP increases CD8+ T-cell frequency and yield in a large cohort of patient samples (n = 34) Percent of CD8+ Cells in TIL Total CD8+ Cells Total TIL Cells Chacon JA, Wu RC, Sukhumalchandra P, Molldrem JJ, et al. (2013) Co Stimulation through 4 1BB/CD137 Improves the Expansion and Function of CD8+ Melanoma Tumor Infiltrating Lymphocytes for Adoptive T Cell Therapy. PLoS ONE 8(4): e doi: /journal.pone
19 Addition of anti-4-1bb antibody to the REP led to increased post- REP TIL tumor antigen-specific CTL activity Chacon JA, Wu RC, Sukhumalchandra P, Molldrem JJ, et al. (2013) Co Stimulation through 4 1BB/CD137 Improves the Expansion and Function of CD8+ Melanoma Tumor Infiltrating Lymphocytes for Adoptive T Cell Therapy. PLoS ONE 8(4): e doi: /journal.pone
20 K562 Cells as Feeder Cells for T cell Expansion K562-Derived Artificial APCs Leukemia cell line Engineered to express Co-stimulatory molecules CD86 and 4-1BBL (CD137 ligand) Membrane-bond IL-15 High affinity Fc receptor CD64 CD86 4-1BBL mil-15 CD64
21 APC-Expanded TIL Contain More CD8+ T Cells Than Those Expanded with PBSC Feeder Cells %CD3+ Cells %CD8+ Cells %CD4+ Cells Forget, Marie-Andree; Malu, Shruti; Liu, Hui; Toth, Christopher; Maiti, Sourindra; Kale, Charuta; Haymaker, Cara; Bernatchez, Chantale; Huls, Helen; Wang, Ena; Marincola, Francesco; Hwu, Patrick; Cooper, Laurence; Radvanyi, Laszlo. Journal of Immunotherapy. 37(9): , November/December Advantages of the Use of APCs as Feeder Cells Better final phenotype product Off the shelf reagent More consistent product 2
22 Producing Large Numbers of Antigen Specific T Cells
23 Melanoma and Colorectal Cancers have Large numbers of Somatic Mutations MS Lawrence et al. Nature 000, 1 5 (2013) doi: /nature12213
24 Identification of Somatic Mutations That Induce T Cell Reponses Tumor Sample Whole Genome Sequencing Culture Tumor Fragments Somatic Mutations Peptide Pools Mini Genes TIL Clones Antigen Presenting Cells Peripheral Blood Monocytes TIL/APC Co- Culture Identify T Cell Responses and Reactive Genes or Peptides Isolate and expand T cells or Isolate TCR for Insertion into autologous T cells
25 CD4+ T helper (T H 1) Cells Recognize a mutation in erbb2 interacting protein (ERBB2IP): Adoptive transfer of TIL containing 25% mutationspecific T H 1 cell stabilize metastatic cholangiocarcinoma Published by AAAS E Tran et al. Science 2014;344:
26 CAR T Cells Directed to CD19 are Very Effective for Treating Acute Lymphocytic Leukemia
27 Genetic Engineering of T cells: High Affinity T Cell Receptors (TCRs) and Chimeric Antigen Receptor (CAR) T Cells High Affinity TCRs CAR Michael Kalos, Carl H. June Immunity Volume 39, Issue
28 Production of Autologous CAR T Cells CD3/CD28 Dynabeads CD19 CAR + Vector 1 st Transduction CD3 + activated T Cells OriGen PermaLife Bags Aphaeresis Peripheral Blood Mononuclear cells ClinExVivo MPC Selection of CD3+ Cells Stimulation of T cell Expansion CD19 CAR + transduced T Cells Final product CD3/CD28 Dynabeads ClinExVivo MPC CD19 CAR + Dynabead Removal Expanded T Cells OriGen PermaLife Bags = CD19 CAR T Cell = Untransduced T Cell 2 nd Transduction Tumaini B, Lee DW, Lin T, Castiello L, Stroncek DF, Mackall C, Wayne A, Sabatino M. Cytotherapy Nov;15(11):
29 Results of Treating 21 Pediatric ALL Patients with Anti-CD19 CAR T Cells by the Pediatric Oncology Branch, NCI, NIH 14 CRs Lee DW et al, Lancet 2014, Oct 10.
30 Anti-CD19 T Cells Persist for Approximately 28 Days Percentage of T cells Expressing CAR Absolute Number of CAR T Cells Number of Copies of CAR Gene Lee DW et al, Lancet 2014, Oct 10
31 Greater Numbers of Circulating Anti-CD19 CAR T Cells is Associated with Clinical Responses Maximum Circulating CAR T Cells Maximum Copies of CAR T Gene Lee DW et al, Lancet 2014, Oct 10
32 CD19 CAR T Cells are Associated with 3 Types of Toxicities Tumor lysis syndrome Cytokine release syndrome Neurotoxicity
33 Cytokine Release Syndrome (CRS) in more likely to Occur in Patients with Great Numbers of Circulating CAR T cells and Greater Tumor Burden Maximum Circulating CAR T Cells Percent Marrow Blasts at Enrollment Lee DW et al, Lancet 2014, Oct 10
34 IL-6 and IFN Levels are Associated with Grade 3 or 4 Toxicity IL-6 IFN- CRS CRS Lee DW et al, Lancet 2014, Oct 10
35 C-Reactive Protein Levels are Associated with IL-6 Levels and Toxicity Relationship between IL-6 and CRP levels in 2 Patients Maximum CPR vs Maximun IL-6 CRP Levels and CRS Lee DW et al, Lancet 2014, Oct 10
36 Standardizing the Assessment and Treatment of CRS Blood July 2014
37 Improving Adoptive Cell Therapy by Using Central Memory or Stem Memory T Cells Restifo et al. Nature Reviews Immunology 2012; 12:
38 Production of Memory Stem T Cells (TSCM) From Peripheral Blood Leukocytes Apheresis Peripheral Blood Mononuclear Cells Anti-CD3 Selection CD3+ Anti-CD62L Selection Anti-CD45RA Depletion CD3+ CD62L+ CD62L+ CD45RA- TCM Cells
39 Cell Selection/Depletion CD3+ Cell CD3 Antibody Binding Selection of CD3+ cells N S Release of CD3+ cells by turning off the magnet N S
40 New Cell Selection Method: Low affinity binding Fab and Streptamers Selected Cell Streptamer-Fab Biotin Releases Streptamers Low Affinity Fabs are Released Stage Biotechnology
41 Conclusions Potency Cell phenotype and specificity are important factors In vivo cell persistency and expansion are associated with increased potency and toxicity More potent cells are in some cases more toxic Manufacturing Manufacturing methods are being modified to produce cells with more desirable phenotypes T cell products with high concentrations of tumor-specific are being manufactured
42 Thank you for your attention!
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