Slide 1: Safety and efficacy of multiTAA-T cells for Myeloma
Safety and efficacy of multi - TAA - T cells for Myeloma Premal Lulla , Ifigeneia Tzannou , George Carrum , Carlos A . Ramos, Rammurti Kamble , Mrinalini Bilgi , Adrian P . Gee Shivani Mukhi , Betty Chung, Ayumi Watanabe, Manik Kuvalekar , Bambi Grilley , Malcolm K . Brenner, Helen E . Heslop , Juan
Slide 2: Problems with myeloma therapy
Therapy Problems Dexamethasone Infections, osteoporosis Melphalan Immunosuppression, second cancers Thalidomide Neuropathy , Clots, anemia Lenalidomide Clots, anemia, second cancers Bortezomib Neuropathy, viral infections ASCT Immunosuppression, infections Problems with myeloma therapy New therapies
Slide 3: MultiTAA T cell therapy for MM
MultiTAA T cells MultiTAA T cell therapy for MM MAGEA4 PRAME Survivin NYESO1 SSX2
Slide 4: Our approach
• Simultaneously target multiple TAAs • Target multiple epitopes (CD4 and CD8) within each antigen • T cells with native T cell receptor specificity (non - engineered) Our approach
Slide 5: MultiTAA-T Cell Generation
MultiTAA - T Cell Generation Activation DC Overlapping pepmixes PBMCs MultiTAA T cells Expansion
Slide 6: Profile of MultiTAA-T cells
n=20 0% 20% 40% 60% 80% 100% CD3+ CD4+ CD8+ DR+/ CD83+ Profile of MultiTAA - T cells RO+/ 62L+ RO+/ 62L - -10% 0% 10% 20% 30% 40% 50% % Positive cells Phenotype Safety % Specific lysis
Slide 7: MultiTAA T cell specificity/polyclonality
1 10 100 1000 MultiTAA T cell specificity/ polyclonality SFC/2x10 5 PRAME NYESO1 SSX2 MA4 Survivin 1 10 100 1000 10000 100000 # of clones mean = 4,597 clones n=10 Clonal diversity (v deep sequencing)
Slide 8: Clinical trial design - Dose escalation
Clinical trial design - Dose escalation (ARM A and B) PRAME/SSX/MAGE/NYESO1/ Survivin - specific T cells : 2 - 4 pts at each level, 2 infusions 14 days apart Dose Level 1 : Day 0 and 14: 5x10 6 cells/m 2 Dose Level 2 : Day 0 and 14: 1x10 7 cells/m 2 Dose Level 3 : Day 0 and 14: 2x10 7 cells/m 2
Slide 9: Clinical Trial - Eligibility
• Any patient > 18 yrs with myeloma diagnosis (post completion of at least 1 treatment regimen) Group A: >90 days post autologous or syngeneic transplant Group B: <90 days post autologous or syngeneic transplant Clinical Trial - Eligibility • No lymphodepletion Group B: <90 days post autologous or s
Slide 10: Group A: Patients Enrolled
ID Age/G Disease DL Prior Treatments 1 53/M IgG - kappa 1 Bor / Dex ASCT 6 61/M IgG - kappa 1 RVD ASCT 7 44/M IgG - kappa 1 CyBorD ASCT 14 47/M IgG - kappa 2 RVD ASCT 3* 65/F IgG - kappa 1 RVD ASCT CyBorD Carf /D ASCT 13 31/F IgG - kappa 2 VD 10 69/F IgG - kappa 2 VD ASCT R Pom
Slide 11: Group A: Patients Infused
ID Age/G Disease Marrow Prior Treatments 1 53/M Active 10% Bor / Dex ASCT 6 61/M In remission 0% RVD ASCT 7 44/M In remission 0% CyBorD ASCT 14 47/M Active 0% (MRD+) RVD ASCT 3* 65/F Active 90% RVD ASCT CyBorD Carf /D ASCT 13 31/F Active 4% VD 10 69/F Active 10% VD ASCT R Pom /
Slide 12: Group B: Patients Enrolled
ID Age/G Disease DL Prior Treatments 2 40/M Free lambda 1 RVD ASCT Pom / Carf /D ASCT 3 65/F IgG - kappa 1 RVD ASCT CyBorD Carf /D ASCT 5 76/M IgG - kappa 1 CyBorD ASCT 8 57/M IgA - kappa 2 VTD ASCT Rd Cy/ Carf /D ASCT 9 50/F IgG - kappa 2 RVD ASCT 11 53/M IgG - lambda 2 VD
Slide 13: Group B: Patients Infused
ID Age/G Disease Marrow Prior Treatments 2 40/M Active 20% RVD ASCT Pom / Carf /D ASCT 3 65/F Active 15% RVD ASCT CyBorD Carf /D ASCT 5 76/M Active 20% CyBorD ASCT 8 57/M In remission 0% VTD ASCT Rd Cy/ Carf /D ASCT 9 50/F In remission 0% RVD ASCT 11 53/M In remission 0% VD
Slide 14: Active Disease: Clinical Outcomes
Active Disease: Clinical Outcomes ID Age/G Disease Marrow Week 6 Wk 6 Mo12 1 53/M Active 10% Unknown SD PR 14 47/M Active 0% (MRD+) 0% (MRD+) SD SD 3* 65/F Active 90% 85% SD PD (2m) 13 31/F Active 4% 0% SD SD 10 69/F Active 10% 10% SD PD (7m) 15 70/M Active 80% 80% SD PD (3m) 2* 40/M Active 15% 15%
Slide 15: In remission: Clinical Outcomes
In remission: Clinical Outcomes Only one patient has relapsed at a median f/u of 21 months ID Age/G Disease Marrow Week 6 Wk 6 Mo12 8 57/M In remission 0% 0% CCR CCR 9 50/F In remission 0% 0% CCR CCR 11 53/M In remission 0% 0% CCR Relapse (7m) 12 54/M In remission 0% 0% CCR CCR 6 61/M In remission 0
Slide 16: Correlating clinical benefit with infused multiTAA T cells
Correlating clinical benefit with infused multiTAA T cells
Slide 17: How can we track non-gene-modified multiTAA T cells in vivo?
How can we track non - gene - modified multiTAA T cells in vivo? Rationale: • In PBMCs (pre - infusion) tumor - specific T cell frequency v. low • below TCR v deep sequencing detection threshold (1/100,000) • Tumor - directed clones enriched in multiTAA T cells • Detectable by v deep sequencing
Slide 18: How many trackable clones are present in our multiTAA T cells?
How many “trackable” clones are present in our multiTAA T cells? 0 20 40 60 80 100 % unique clones 1 10 100 1000 10000 100000 # of clones mean = 4,597 clones n=10 Clonal diversity in multiTAA T cells Unique clones 59% 95% Avg = 80% Pt#2
Slide 19: What drives in vivo multiTAA expansion?
• Patients enrolled on different arms depending on proximity to transplant [> ( Grp A ) or < ( Grp B ) 90 days] • Does post - transplant lymphodepletion impact expansion? What drives in vivo multiTAA expansion? • Patients with and without disease enrolled on study • Does presence of antigen influenc
Slide 20: Antigen drives multiTAA expansion - TCR tracking
0 0.2 0.4 0.6 0.8 1 1.2 Pre Early Late Antigen drives multiTAA expansion – TCR tracking 0 0.2 0.4 0.6 0.8 1 1.2 1 Production frequency Peripheral blood Marrow Post infusion Production frequency Post infusion Active disease No disease
Slide 21: Antigen drives multiTAA expansion - ELIspot
0 10 20 30 40 50 60 70 80 Preinf Week 6 Mo 6 Mo 12 Antigen drives multiTAA expansion – ELIspot Active disease No disease SFC/5x10 5 PBMCs Peripheral blood pre wk6 mo6 mo12 0 20 40 60 Pre wk6 mo6 mo12 0 20 40 60 pre wk6 mo6 mo12 Active disease No disease Survivin NYESO1 MAGEA4 SSX2 PRAME SFC/5x10 5 P
Slide 22: T cell kinetics in responders
T cell kinetics in responders
Slide 23: Clinical Response - Pt#2
Clinical Response – Pt#2 20% Clonal plasma cells Diagnosis Pre 1st ASCT ASCT +6m ASCT +22m Pre 2nd ASCT +27m Pre-T cells (day+61) Post-T cells (day+113) Month 6 2 nd line 2 nd ASCT 1 st line 1 st ASCT 3 rd line 100 1000 10000 0 MultiTAA T cells Free lambda mg/L
Slide 24: Clinical Response - Pt#2
Clinical Response – Pt#2 <1% Clonal plasma cells Cyclin D1 neg 20% Clonal plasma cells Diagnosis Pre 1st ASCT ASCT +6m ASCT +22m Pre 2nd ASCT +27m Pre-T cells (day+61) Post-T cells (day+113) Month 6 2 nd line 2 nd ASCT 1 st line 1 st ASCT 3 rd line 100 1000 10000 0 Normal MultiTAA T cells Free lambd
Slide 25: Clinical Response - Pt#2
0 50 100 150 200 250 300 Pre Wk4 Wk6 Mo3 SFC/5x10 5 60 120 180 0 6 Wk Post SFC/5x10 5 MAGE A4 SSX2 Survivin NYESO - 1 PRAME Targeted antigens PBMC Marrow Clinical Response – Pt#2 MAGE A4 SSX2 Survivin NYESO - 1 PRAME
Slide 26: Clinical Response - Pt#2
60 120 180 0 6 Wk Post SFC/5x10 5 MAGE A4 SSX2 Survivin NYESO - 1 PRAME Marrow Clinical Response – Pt#2 MAGE - A4 3+ Pre - infusion 1+ 8wk post T cells
Slide 27: In vivo T cell tracking - Pt#2
Targeted antigens SFC/5x10 5 0 50 100 150 200 250 300 Pre Wk4 Wk6 Mo3 60 120 180 0 6 Wk Post MAGE A4 SSX2 Survivin NYESO - 1 PRAME MAGE A4 SSX2 Survivin NYESO - 1 PRAME SFC/5x10 5 PBMC Marrow In vivo T cell tracking – Pt#2 0 0.1 0.2 0.3 0.4 0.5 Pre Wk4 Mo6 % Production Frequency 0 0.1 0.2 0.3 Wk 6 P
Slide 28: Immune escape post multiTAA T cells
Immune escape post multiTAA T cells
Slide 29: Clinical Course - Pt#3
0 1 2 3 4 5 6 7 8 Diagnosis 1st line PD+3y Pre 1st HSCT HSCT +2y Pre 2nd HSCT Pre - T cells (d+47) Post - T cells (d+89) IgG kappa M - spike (g/dl) Clinical Course - Pt#3 2 nd line 3 rd line 1 st HSCT 2 nd HSCT 1 st line MultiTAA T cells 15% Clonal plasma cells
Slide 30: Clinical Course - Pt#3
0 1 2 3 4 5 6 7 8 Diagnosis 1st line PD+3y Pre 1st HSCT HSCT +2y Pre 2nd HSCT Pre - T cells (d+47) Post - T cells (d+89) IgG kappa M - spike (g/dl) Clinical Course - Pt#3 2 nd line 3 rd line 1 st HSCT 2 nd HSCT 1 st line 15% Clonal plasma cells 10% Clonal plasma cells MultiTAA T cells
Slide 31: Clinical Course - Pt#3 - ELIspot
0 20 40 60 80 100 Pre Wk4 Wk6 Mo3 SFC/2x10 5 MART1 MAGE C1 WT1 AFP MAGE A3 MAGE A2B MAGE A1 Non - targeted antigens 0 20 40 60 80 100 120 Pre Wk4 Wk6 Mo3 SFC/5x10 5 MAGE A4 SSX2 Survivin NYESO - 1 PRAME Targeted antigens PBMC SFC/5x10 5 PRAME Marrow 0 200 400 600 6 wk POST Clinical Course - Pt#3 - E
Slide 32: Clinical Course - Pt#3
0 20 40 60 80 100 Pre Wk4 Wk6 Mo3 SFC/2x10 5 MART1 MAGE C1 WT1 AFP MAGE A3 MAGE A2B MAGE A1 Non - targeted antigens 0 20 40 60 80 100 120 Pre Wk4 Wk6 Mo3 SFC/5x10 5 MAGE A4 SSX2 Survivin NYESO - 1 PRAME Targeted antigens PBMC SFC/5x10 5 PRAME Marrow 0 200 400 600 6 wk POST Clinical Course - Pt#3 mul
Slide 33: Clinical Course - Pt#3
0 1 2 3 4 5 6 7 8 Diagnosis 1st line PD+3y Pre 1st HSCT HSCT +2y Pre 2nd HSCT Pre - T cells (d+47) Post - T cells (d+89) Month 6 Month 8 Clinical Course - Pt#3 2 nd line 3 rd line 1 st HSCT 2 nd HSCT 1 st line MultiTAA T cells 10% Clonal plasma cells 15% Clonal plasma cells IgG kappa M - spike (g/dl
Slide 34: Clinical Course - Pt#3
0 1 2 3 4 5 6 7 8 Diagnosis 1st line PD+3y Pre 1st HSCT HSCT +2y Pre 2nd HSCT Pre - T cells (d+47) Post - T cells (d+89) Month 6 Month 8 Clinical Course - Pt#3 2 nd line 3 rd line 1 st HSCT 2 nd HSCT 1 st line 90% Clonal plasma cells 10% Clonal plasma cells 15% Clonal plasma cells IgG kappa M - spik
Slide 35: Clinical Course - Pt#3
0 20 40 60 80 100 Pre Wk4 Wk6 Mo3 Mo6 SFC/2x10 5 MART1 MAGE C1 WT1 AFP MAGE A3 MAGE A2B MAGE A1 Non - targeted antigens 0 20 40 60 80 100 120 Pre Wk4 Wk6 Mo3 Mo6 SFC/5x10 5 MAGE A4 SSX2 Survivin NYESO - 1 PRAME Targeted antigens PBMC 6 mo POST SFC/5x10 5 Marrow 0 200 400 600 6 wk POST MAGE A4 SSX2 S
Slide 36: Mechanism of Escape
Pre Wk8 mth6 50 40 30 20 10 0 Pre Wk8 mth6 80 60 40 20 0 SFC/5x10 5 Pre Wk8 mth6 20 0 15 10 5 Pre Wk8 mth6 100 80 60 40 20 0 2 1 0 IHC Score MAGE - A4 2+ PRAME 2+ Survivin 1+ NYESO1 neg Pre Mechanism of Escape
Slide 37: Mechanism of Escape
Pre Wk8 mth6 SFC/5x10 5 100 80 60 40 20 0 Pre Wk8 mth6 50 40 30 20 10 0 2 1 0 Pre Wk8 mth6 80 60 40 20 0 Pre Wk8 mth6 20 0 15 10 5 IHC Score MAGE - A4 2+ PRAME 2+ Survivin 1+ NYESO1 neg Pre Mechanism of Escape
Slide 38: Mechanism of Escape
Pre Wk8 mth6 SFC/5x10 5 100 80 60 40 20 0 Pre Wk8 mth6 50 40 30 20 10 0 2 1 0 Pre Wk8 mth6 80 60 40 20 0 Pre Wk8 mth6 20 0 15 10 5 IHC Score 0 20 40 60 80 100 120 Pre Wk4 Wk6 Mo3 Mo6 SFC/5x10 5 MAGE A4 SSX2 Survivin NYESO - 1 PRAME Targeted antigens MAGE - A4 PRAME Survivin NYESO1 Mechanism of Escap
Slide 39: Mechanism of Escape
variable genes (n=1,828) MS4A1 IL1B CD86 CTLA4 LAG3 Pre wk6 mo3 mo6 IL6 PD1 SD mo3 MM cell proliferation (n=407) Fulciniti M et al, Blood Cancer J, 2016 Mitchell JS et al, Nat commun , 2016 Mechanism of Escape Pre wk6 mo3 mo6 SD PD SD PD Immune activating genes Immune inhibitory genes PD mo6 Linghua
Slide 40: MultiTAA T cells for myeloma
• Safe to date (DL3 – Arm A & B) • Feasible • In vivo expansion of tumor - specific T cells directed to target antigens • Antigen spreading • Clinical benefit MultiTAA T cells for myeloma
Slide 41: Acknowledgements
TRL Lab PIs Helen Heslop Cliona Rooney Malcolm Brenner Juan Vera Ann Leen QA/QC Laboratory Adrian Gee Sara Richman Natasha Lapteva Debbie Lyon April Durett Suzanne Poole Zhuyong Mei Crystal Silva - Lentz GMP Laboratory Huimin Zhang Birju Mehta Funding : Leukemia Texas Reseach grant, Leukemia and Lym