CAR T CELLS - A PERSONALIZED ATTACK ON CANCER. Xiuli Wang, PhD

Transcription

1 CAR T CELLS - A PERSONALIZED ATTACK ON CANCER Xiuli Wang, PhD

2 Disclosures I do not have anything to disclose

3 Evolution of CAR T Cell Therapy CD28 41BB Levine et al Cancer Gene Therapy 2015 Mackall et al 2014

4 CD19: An Ideal Target for Immunotherapy Blanc et al. Clin Cancer Res :6448

5 FDA Approval for CD19-CAR T Therapy KYMRIAH August 30, 2017 Novartis Receives Approval For Pediatric Acute Lymphoblastic Leukemia KYMRIAH May 01, 2018 Large B-Cell Lymphoma YESCARTA October 18, 2017 Kite/Gilead Receives Approval For Adult Non-Hodgkin Lymphoma

6 Building Better and New CAR T Cell Therapy

7 Variables Required for Effective CAR T Cell Therapy Starting population: unselected PBMC, memory subsets, defined CD4/CD8 Activation: CD3/CD28, OKT3 Ex vivo expansion: cytokines CAR design: scfv, spacer Clinical management Priceman et al COO, 2015

8 The T Cell Compartment Consists of T Cell Subsets at Different Stages of Differentiation Naïve (T N ) Memory Stem Cell (T SCM ) Central Memory (T CM ) Effector Memory (T EM ) Effector (T E ) Longevity Self-renewal Proliferative potential Memory cell potential Cytotoxicity INFγ production Short lived Gattinoni et al. Nat Rev. Cancer 2012

9 T CM Derived Human Effectors Exhibit Superior Persistence Anti-tumor Activity Following Adoptive Transfer RO bleeds Wang et al Blood, 2011 Wang et al Oncoimmunology, 2015

10 CD19R(EQ)28z/EGFRt+ T in cgmp Vuelife Bag 1 - Leukapheresis CD3/CD28 Stimulation Lentitransduce with CD19R(EQ)28Z-T2A-EGFRt_epHIV Lentiviral Transduction Initiate Expansion 2 - CliniMACS Selection; Option to Freeze Cells; Cryopreservation 7+ - Dynabead Removal TM Day - Bioprocess: Platform Development for the Generation of CAR T Cells from Defined T Cell Subsets for Clinical Trial Deplete Deplete1) Negative selection Deplete CD25+ CD25+ CD25+ CD14+ CD14+ CD14+ CD45RA+ CD45RA+ CD45RA+ PBMC CM PBMCs CD25+ Deplete CD45RA+ CD14+ using biotin-dreg56 CD62L+ for Select and anti-biotin microbeads CD62L+ T CM CD3/CD28 with Stimulate Dynabeads with Expand IL-2/IL-15 PBMC PBMC CD25- CD14- CD45RA- Deplete CD25+ CD14+ CD25- CD25- CD45RA+ CD14- CD14- CD45RA- CD45RA- Day - Bioprocess: Day - Bioprocess: Day - Bioprocess: 1 - Leukapheresis PBMC 1 - Leukapheresis 1 - Leukapheresis Day - Bioprocess: TM TM 2 - CliniMACS Selection; 2 - CliniMACS Selection; TM PBMC 1 - Leukapheresis 2 - CliniMACS Option to Freeze Cells; Option to Freeze Selection; Cells; Option CD3/CD28 Stimulation CD3/CD28 to Freeze Stimulation Cells; TM 2 - CliniMACS Selection; CD3/CD28 Stimulation CD25- Option to Freeze Cells; CD14- CD3/CD28 Stimulation CD45RA- CD25- CD14- CD45RA- Select for CD62L+ Select for CD62L+ Select 2) Positive for CD62L+ selection using biotin-dreg56 using biotin-dreg56 CD62L+ Memory T cells using biotin-dreg56and anti-biotin microbeads and anti-biotin microbeads Select for CD62L+ and anti-biotin microbeads using biotin-dreg56 and anti-biotin microbeads 3) Stimulate with CD62L+ T CD62L+ T CM CD3/CD28 Dynabeads CD62L+ T CM Stimulate CM with Stimulate with Stimulate with CD3/CD28 CD62L+ T CD3/CD28 CM CD3/CD28 Dynabeads Dynabeads Stimulate with Dynabeads 4) Lentitransduce with CD3/CD28 CD19R(EQ)28ζ-T2A-EGFRt_epHIV7 Dynabeads Lentitransduce with Lentitransduce with Lentiviral Transduction; Lentitransduce CD19R(EQ)28Z-T2A-EGFRt_epHIV7 with Lentiviral Transduction; Initiate Expansion 5) Expand with CD19R(EQ)28Z-T2A-EGFRt_epHIV Lentiviral Transduction; CD19R(EQ)28Z-T2A-EGFRt_epHIV7 IL-2/IL-15 Initiate Expansion Expand Lentitransduce Initiate Expansion with Lentiviral Transduction; Expand Expand withcd19r(eq)28z-t2a-egfrt_ephiv Dynabead Initiate Expansion with Removal 6) CD19R(EQ)28ζ-T2A-EGFRt+ with IL-2/IL Dynabead Removal IL-2/IL Dynabead Expand Removal Tcm or Tn/scm/cm IL-2/IL-15 with 7+ - Dynabead Removal in cgmp Vuelife bag CD19R(EQ)28z/EGFRt+ IL-2/IL-15 TCM CD19R(EQ)28z/EGFRt+ T Cryopreservation CD19R(EQ)28z/EGFRt+ in cgmp T CMVuelife Bag Wang et al, J Immunother Cryopreservation in cgmp Vuelife Bag CM Cryopreservation in cgmp Vuelife Bag CD19R(EQ)28z/EGFRt+ TCM Cryopreservation in cgmp Vuelife Bag

11 IRB13447 Therapeutic Activity of CD19-CAR T CM (CD19R (EQ)28z EGFRt) for Patient with ALL UPN yr; female Relapse ALL; BCR-ABL positive Post 3x hyper-cvad (Cytoxan) chemotherapy; Imatinib/Dasatinib/Ponatinib; 2x Allo-transplant (double cord blood);blinitumomab Treated with CD19-CAR T cells - Day -3: 1.5 gm/m2 Cytoxan; 2 days - Day 0: 200M CD19-CAR T CM

12 Therapeutic Activity of CD19-CAR T CM (CD19R (EQ)28z EGFRt) for Patient with ALL Anti-leukemic Activity of CD19CAR T Cells In vivo Persistence of CD19CAR T Cells

13 The T Cell Compartment Consists of T Cell Subsets at Different Stages of Differentiation Naïve (T N ) Memory Stem Cell (T SCM ) Central Memory (T CM ) Effector Memory (T EM ) Effector (T E ) Longevity Self-renewal Proliferative potential Memory cell potential Cytotoxicity INFγ production Short lived Gattinoni et al. Nat Rev. Cancer 2012

14 S u p B 1 5 -E G F P :ffl u c F lu x (p h o to n s /s e c ) Naïve/Memory-Derived CAR T Cells Show Improved Antitumor Efficacy Day 23 SupB15 CD19+ ALL S u p B 1 5 P B M C M o c k P B M C -C A R T C M -C A R T N / M E M -C A R PBMC-mock PBMC-CAR T CM -CAR D a y p o s t T c e ll in je c tio n T N/MEM -CAR

15 % C A R T c e l l s % C A R T c e l l s Tn/mem-derived CAR T Cell Exhibit Improved Expansion Pre-T cell Day 7 Day 14 Day 21 Day 28 Tcm CAR (UPN084) 0.4% 1.9% 0.8% 0.3% 0 Tn/mem CAR (UPN137) 0.3% 23.4% 31.4% 6.9% 1.1% Tcm-derived CD19-CAR T cells (200M) U P N U P N U P N Tn/mem-derived CD19-CAR T cells (200M) U P N U P N U P N U P N U P N U P N ( P r e ) D a y P o s t - i n f u s i o n 0 0 ( P r e ) D a y P o s t - i n f u s i o n Clinical trials: Dr. Leslie Popplewell Dr. Samer Khaled

16 CNS Lymphoma and Treatment Primary CNS Lymphoma (PCNSL) Tumor from lymph tissue starts in the brain, spinal cord, and/or meninges 1900 new cases per year in U.S. The incidence has been increasing over the past 20 years It represents between 2% and 3% of all primary brain tumors. Unlike peripheral lymphomas, PCNSL is seen exclusively in advanced disease Solitary lesions in 50% and multicentric lesions in 50% Deep lesions are common 13% 44% 28% 14% 6% 6% 6% <1% Multiple lesions Deep lesions Fallah et al., Blood Adv, 2016

17 CAR T Cell Therapy for CNS Lymphoma Brown et al, N Engl J Med, 2016 Priceman et al, Clin Cancer Res 2017 Intracerebroventricular (i.c.v) CAR T Cell Therapy Feasible Safe Highly effective We want to evaluate and compare the efficacy of intracerebroventricularly (i.c.v) and intravenously (i.v.) delivered CAR T cells against CNS lymphoma

18 CD8 CD8 APC-Cy7-A CD8 APC-Cy7-A CD8 i.c.v. Delivered CD19-CAR T Cells Exhibit Superior Anti-CNS Lymphoma Activity Untreated CNSL (Daudi) i.c.v. Tumor Mock i.c.v % 27.27% 5 10 CD19CAR 1.2x10^ i.v. i.c.v.: Intracerebroventricular Mock i.v. CD19CAR 3.0x % 18.97% CAR EGFR APC-A 6.96% 5.70% % 30.70% CAR EGFR APC-A CAR T cells migrate from the brain to periphery blood Can i.c.v. CAR control systemic lymphoma?

19 P e rc e n t s u rv iv a l i.c.v. Delivered CD19-CAR T Cells Completely Eradiate Brain and Systemic Lymphoma IV 2x10^6 CD19-CAR T cells ICV Untreated mock endpoint * Tumor irrelevant death i.c.v. U n tre a te d CD19-CAR T 5 0 P<0.001 M o c k i.c.v. C D 1 9 C A R i.c.v. Daudi lymphoma s.c. & i.c. mock i.v. CD19-CAR T M o c k i.v. C D 1 9 C A R i.v. D a y s p o s t T c e ll in fu s io n Potential application: Primary CNS lymphoma and B cell malignancies with CNS involvement Clinical trials: Dr. Tanya Siddiqi Dr. Jana Portnow

20 Multiple Myeloma (MM) Multiple myeloma (MM), a plasma cell malignancy, accounts for slightly more than 10 percent of hematologic malignancies in the United States. Estimated new cases in 2015 are 26,850; Current therapies for MM often induce remission, 5 year survival rate 46.6%: ; nearly all patients eventually relapse. Over 11,000 people die from this disease/year. T-cell mediated anti-tumor therapies using genetically modify T cells with specific chimeric antigen receptors (CARs) have non-overlapping activity, toxicity and tumor resistance profiles compared to conventional chemotherapeutic agents.

21 CS1 (SLAMF7) CS1 is a cell surface glycoprotein of SLAM receptor family CS1 is uniformly expressed on plasma cells and in over 95% cases of MM, irrespective of cytogenetic abnormalities and throughout the disease process Low expression on T cells, NK cells and no expression on HSC and other normal tissues

22 % p o s itiv e c e lls CS1 (SLAMF7): A Target for T Cell Therapy of Multiple Myeloma (MM) Multiple Myeloma Cells are CS1 Positive C S 1 s u r fa c e a n tig e n MM cells T cells NK cells CS1 scfv-fc

23 P e rc e n t s u rv iv a l huigg4 hinge-fc CS-1 CAR Re-directed T Cell Immunotherapy for Multiple Myeloma CS1 CAR hucd4tm hucd3-z cyto Day Untreated Mock U n tre a te d M o c k C S 1 C A R P=0.003 CS1 CAR D a y s p o s t T c e ll in fu s io n Wang et al Clin Cancer Res Clinical vector:

24 CS1 CAR T Cell Therapy for Multiple Myeloma Deplete Body s Immune cells Dose level -1 Dose level 0 (Starting dose) Dose level 1 Dose level 2 Cell Dose million million million million CAR DNA 06/15 GMP CAR 11/15 CAR Cert. of Analysis 04/16 FDA proceed 11/18 Feb Clinical trials: Dr. Myo Htut

25 Mutilfunctional CAR T Cells for More Controllable T Cell Therapy Tumor targeting Expansion Ablation/selection/tracking CAR T cells are ablated in vivo pp65xcmv bi-specific T cells with EGFR for selection/ablation Wang et al Blood 2011 Bispecific T cells CMV specific T cells CMVxCD19CAR CMVxCD19 CAR CMVxCD19 CAR CMVxCD19 CAR CMVxCD19 CAR CMV vaccine Augmented anti-tumor activity

26 Improved Anti-lymphoma Efficacy of Adoptively Transferred CMV-CD19CAR Bi-specific T Cells by CMV Vaccine CD19 + ffluc + tumor CMVxCD19CAR T cells pp65 vaccination (pp65peptide pulsed T cells) Day: Untreated Euthanized CMV-specific T pp65 vaccine Euthanized Prior Vaccine Post CMVxCD19CAR T pp65 vaccine CMVxCD19CAR T mp1 vaccine Wang et al Clin Cancer Res Clinical trials: Dr. Ryotaro Nakamura Lymphoma SPORE

27 % C A R (o f to ta l h u C D c e lls ) % C y to to x ic ity CD8 APC-Vio770-A CMV-HIV CAR T Cell Therapy for Patients with HIV/AIDS HIV gp Response to HIV (gp120) C M V x N 6 C M V LL_CMVxN6_v pp65+ PBMC_C fcs CD % 20.06% Response to CMV :1 1 :1 1 :5 E :T R a tio % 67.45% IFNg APC-A * * * * * B e fo r e V a c c in a tio n A fte r V a c c in a tio n Clinical trials: Dr. Elizabeth Budde

28 Future Directions : Building a New and Better CAR T Cell Therapy Increase ratio of efficacy/toxicity Suboptimal potency CRS/ Neurotoxicity Durability of response On target off tumor effects Antigen escape Immunosuppressive tumor microenvironments

29 Acknowledgements Stephen Forman, MD TCTRL Research Xiuli Wang team: Laura Lim Christian Huynh Vibhuti Vyas TCTRL Translational Team Jamie Wagner and Team Araceli Naranjo and Team Wen-Chung Chang and Team Jinny Paul and Team Julie Ostberg Kirsten Rood James Sanchez Jennifer Shepphird Achini Bandara Sandra Thomas Hematologic Malignancy Amrita Krishnan Myo Htut Michael Rosenzweig Leslie Popplewell Elizabeth Budde Khaled Tanya Siddiqi Larry Kwak Hong Qin Ryo Nakamura John Zaia Don Diamond Angelo Cardoso Kevin Morris John Burnett Samer Joseph Cohen Ryan Urak Miriam Water Christine brown and Team Saul Priceman and Team Elizabeth Budde and Team Vanessa Jonsson and Team Larry Stern Zhiqiang Wang All Forman Members Funding Agencies NCI Lymphoma SPORE SCOR Mantle Cell Lymphoma Research Initiative Award CIRM Borstein Family Foundation Steven Gordon & Briskin Family Innovation Grant MM Research Foundation AL Amyloid Foundation