Adoptive Cell Therapy: Treating Cancer with Genetically Engineered - PowerPoint PPT Presentation

Adoptive Cell Therapy: Treating Cancer with Genetically Engineered T Cells ith G ti ll E i d T C ll Steven A. Feldman, Ph.D. Surgery Branch S B h National Cancer Institute NCT Conference Heidelberg, Germany September 24 2013 September 24, 2013

Three Main Approaches to Cancer Immunotherapy 1. Non-specific stimulation of immune reactions • Stimulate effector cells (IL-2, IL-12) • • Inhibit regulatory factors (PD 1 CTLA 4) Inhibit regulatory factors (PD-1, CTLA-4) 2. Active immunizations to enhance anti-tumor reactions • Cancer vaccines 3. Passively transfer activated immune cells with anti-tumor 3 P i l f i d i ll i h i activity • • Adoptive cell transfer Adoptive cell transfer

Advantages of Cell Transfer Therapy Advantages of Cell Transfer Therapy 1. High avidity anti-tumor T cell receptors (TCR) can be identified and cloned using in vitro assays. 2. Peripheral blood lymphocytes can be genetically modified to express these high avidity TCRs. p g y 3. Large numbers of tumor-specific lymphocytes can be grown in vitro. i i 4 4. The host can be manipulated to provide a favorable tumor The host can be manipulated to provide a favorable tumor microenvironment prior to administering the cells. 5. ACT can mediate tumor regressions.

Development of Adoptive Cell Transfer Therapy

A Critical Challenge Confronting the Development of Human Cancer Immunotherapy is the of Human Cancer Immunotherapy is the Identification of Antigens to Target 1. Differentiation antigens overexpressed on cancers compared to normal tissue (MART-1, gp100, CEA, Her-2, Mesothelin) 2. 2 Antigens expressed on cancers and on non essential normal Antigens expressed on cancers and on non-essential normal tissues (CD19, thyroglobulin) 3. Shared antigens unique to cancer (cancer-testes antigens, NY- ESO-1, MAGE-A) 4. Mutations unique to each cancer (EGFRvIII) 5. Critical components of the tumor stroma (VEGFR2, FAP)

Surgery Branch Gene Therapy Products (by class) (by class) IL 2 IL-2 Cytokine IL 12 IL-12 murine (gp100, NY-ESO-1, MAGE-A3) murine ( 100 NY ESO 1 MAGE A3) TCR human (DMF5 NY ESO 1) human (DMF5, NY-ESO-1) 2 nd gen 28Z (CD19 Meso) 2 nd gen-28Z (CD19, Meso) CAR 3 rd gen 28BBZ (EGFRvIII VEGFR2) 3 rd gen-28BBZ (EGFRvIII, VEGFR2)

Interleukin 12 • IL-12 is a heterodimeric cytokine composed of a heavy chain IL 12 is a heterodimeric cytokine, composed of a heavy chain (p40) and a light chain (p35), Coordinated production of the two chains lead to the secretion of the biologically active p70 • IL-12 is produced by activated hematopoietic phagocytic cells (monocytes, macrophages, neutrophils) and dendritic cells (DC) • Activates effector cells: CD4+, CD8+, and NK cells

Development of an Inducible Vector to Mediate IL-12 Production Only in the Tumor Microenviroment Production Only in the Tumor Microenviroment MSGV1.NFAT.IL12.PA2 In vitro In vivo No IL ‐ 2, no vaccine , Zhang L, Kerkar SP et al, Molecular Therapy, 2011

Phase I Study of ACT Using TIL Transduced with Gene Encoding IL 12 (9/28 32% OR) Gene Encoding IL-12 (9/28, 32% OR) TIL grown for 2-3 weeks Stimulated with OKT-3, transduced and expanded , p Infuse after Cy/flu preparative regimen No IL-2 administered Cohort Number Result (# cells x 10 -9 ) of patients ___________________________________________________________________________ 0.001 1 1NR 0.003 1 1NR 0.01 7 7NR 0.03 0 03 5 5 1CR (24+ mos); 4NR 1CR (24+ mos); 4NR 0.1 3 3NR 0.3 3 3PR (4, 6, 12+) 1.0 4 1PR (12+); 3NR 3 0 3.0 4 4 1CR (5+) 3PR (9+ 7 1CR (5+) 3PR (9+, 7, 5) 5) In first 5 cohorts 1 of 17 patients responded. At doses greater than 0.1X10 -9 , 8 of 11 patients responded.

IL-12 Gene Therapy (M.S. 3x10 7 IL12Td CD8+ TIL)

Summary TIL IL-12 • Tumor infiltrating lymphocytes transduced with NFAT.IL12 vector T i fil i l h d d i h NFAT IL12 secrete IL-12 upon stimulation. • 28 patients with metastatic melanoma received the autologous TILs genetically modified by NFAT.IL12 vector. • Following IL12 Td TILs infusion, 2 patients experienced dose limiting toxicity correlated with high levels of IFN-g and IL-12 in their g y g g serum. All patients recovered completely. • 9 9 out of 28 ( 32%) patients responded to IL-12 Td-TIL treatment t f 28 ( 32%) ti t d d t IL 12 Td TIL t t t based on RECIST. IL-2 not needed to achieve OR in this setting.

Surgery Branch Gene Therapy Products (by class) (by class) IL 2 IL-2 Cytokine IL 12 IL-12 murine (gp100, NY-ESO-1, MAGE-A3) murine ( 100 NY ESO 1 MAGE A3) TCR human (DMF5 NY ESO 1) human (DMF5, NY-ESO-1) 2 nd gen 28Z (CD19 Meso) 2 nd gen-28Z (CD19, Meso) CAR 3 rd gen 28BBZ (EGFRvIII VEGFR2) 3 rd gen-28BBZ (EGFRvIII, VEGFR2)

T-cell Receptor (TCR) Gene Therapy TCR Cloning IVS Immunize mice TCR Vector (eg, MART1, NY ‐ ESO) TCR receptor α β SD SA Ψ LTR TCR α 2A TCR β LTR CD3 ζ , γ , ε , δ

Cancer/Testes Antigens - Shared Tumor Specific Antigens Antigens Expressed during fetal development Expressed during fetal development Restricted in their expression in adult normal tissues to germ cells Up-regulated in 10-80% of cancers from multiple tissues NY-ESO-1 Family Small family of X-linked genes that includes NY-ESO-1 and LAGE-1 MAGE Family Family of ~ 45 X-linked genes

Cancer/Testis Antigens Expressed in Multiple Tumor Types Types 80 R r by RT ‐ PCR 70 60 MAGE ‐ A3 50 50 tive tumor MAGE ‐ A1 40 30 NY ‐ ESO ‐ 1 % of posit 20 10 0 Bladder r C NSCLC Melanoma a n Ovarian r patocellular a Myeloma a ll carcinoma quamous cel He Tumor Type Tumor Type Sq

Recognition of Non-melanoma Tumors by NY- Recognition of Non melanoma Tumors by NY ESO-1 TCR Transduced PBL ESO A2 MSGIN Glioblastoma LNZAT3WT4 - + APB, NY-ESO-1 TCR + + Ewing’s sarcoma TC-71 + + Glioblastoma LN-18 + + + + SCLC SCLC NCI H526 NCI H526 + + Neuroblastoma SKN-AS-A2 + + Breast cancer MDA453s-A2 + + Saos-2 Osteosarcoma + + SCLC NCI H345 Melanoma + + 624.38mel 0 100 200 300 400 500 600 IFN-  , pg/ml IFN pg/ml

Phase II Study of Metastatic Cancer that Phase II Study of Metastatic Cancer that Expresses NY-ESO-1 Using Lymphodepleting Conditioning Followed by Infusion of Anti-NY- Conditioning Followed by Infusion of Anti NY ESO-1 TCR-Gene Engineered Lymphocytes J. Clinical Oncology, 29:917 ‐ 924, 2011

DC (Melanoma) CR 30+

HK (Synovial Cell Sarcoma) PR (14*)

SUMMARY: NY-ESO-1 TCR Engineered T cells TCR gene therapy targeting CTA antigen NY-ESO-1 can lead to cancer regression in melanoma and synovial cell sarcoma without associated g y toxicities. Total PR CR OR Total PR CR OR _______________________________________________________________ number of patients (duration in months) Melanoma 18 5 (28%) 4 (22%) 9 (50%) (18+,10**, 8, 4, 3) (48+, 37+, 25, 21+**) Synovial Cell 16 10 (63%) 0 10 (63%) Sarcoma (29+**,14*, 12**,10, 8, 6+, 5, 4, 3**,2+) *t *treated twice t d t i **plus ALVAC vaccine (Robbins et al J Clin Oncol29:917-924, 2011)

Limitation of TCR gene transfer 1. HLA-restriction limits ability to treat patients / requires multiple TCRs p 2. Inability to target lipid / carbohydrate molecules y g p y 3. 3. Potential tumor “escape” via MHC loss / alterations in antigen Potential tumor escape via MHC loss / alterations in antigen processing

Surgery Branch Gene Therapy Products (by class) (by class) IL 2 IL-2 Cytokine IL 12 IL-12 murine (gp100, NY-ESO-1, MAGE-A3) murine ( 100 NY ESO 1 MAGE A3) TCR human (DMF5 NY ESO 1) human (DMF5, NY-ESO-1) 2 nd gen 28Z (CD19 Meso) 2 nd gen-28Z (CD19, Meso) CAR 3 rd gen 28BBZ (EGFRvIII VEGFR2) 3 rd gen-28BBZ (EGFRvIII, VEGFR2)

Chimeric Antigen Receptors (CARs) Step 2 Step 3 Step 1 Ig ScFv Linker/TM T cell signaling Antibody Producing Hybridoma Ig Genes Chimeric Antigen Receptor (CAR) CAR (CD19, Meso, EGFRvIII, VEGFR2 CAR receptor   sa sa sd sd V L V L scFv LTR LTR CD28 CD3 zeta Anti-tumor Ag-scFv V H V H  CD28 sa sd LTR LTR Anti-tumor Ag-scFv CD8 CD28 4-1BB CD3 zeta CD3 ζ

B-Cell Malignancies (A ti (Antigens Expressed on Non-Essential Normal Tissues) E d N E ti l N l Ti ) • Approximately 22,000 people die of B-cell malignancies Approximately 22 000 people die of B cell malignancies annually in the U.S. • CD19 is expressed by more than 90% of B cell • CD19 is expressed by more than 90% of B-cell malignancies. • CD19 is expressed by mature B cells B-cell precursors CD19 is expressed by mature B cells, B cell precursors and plasma cells but not any other normal tissues. Anti ‐ CD19 CAR  5’ LTR FMC63 scFv CD28 CD3 ‐ zeta 3’ LTR

Bone marrow biopsies showed extensive CLL before and nearly absent B-lineage cells after treatment y g Before treatment 3 months after treatment CD19 CD19 CD20 CD20 Kochenderfer et al. Blood 2012

Tumor regression and elimination of normal B cells