Genetically modified anti-third party central memory T cells and use of same in immunotherapy
An isolated cell having a central memory T-lymphocyte (Tcm) phenotype, the cell being tolerance-inducing cell and capable of homing to the lymph nodes following transplantation, the cell being transduced to express a cell surface receptor comprising a T cell receptor signaling module is disclosed. Methods of generating same and using same are also disclosed.
1. An isolated cell having a central memory T-lymphocyte (Tcm) phenotype, said cell being a non-graft versus host (GVHD)-inducing anti-third party cell, being tolerance-inducing and capable of homing to the lymph nodes following transplantation, said cell being transduced to express a cell surface receptor, said cell surface receptor comprising a T cell receptor signaling module and an extracellular domain directed against a disease antigen.
2. An isolated cell having a central memory T-lymphocyte (Tcm) phenotype, said cell being a non-graft versus host (GVHD)-inducing anti-third party cell, being tolerance-inducing and capable of homing to the lymph nodes following transplantation, said cell being transduced to express a chimeric antigen receptor (CAR) comprising an extracellular domain directed against a disease antigen.
3. An isolated cell having a central memory T-lymphocyte (Tcm) phenotype, said cell being a non-graft versus host (GVHD)-inducing anti-third party cell, being tolerance-inducing and capable of homing to the lymph nodes following transplantation, said cell being transduced to express a chimeric antigen receptor (CAR), wherein said CAR comprises a co-stimulatory domain and comprises an extracellular domain directed against a disease antigen.
4. The isolated cell of claim 1 , wherein said cell surface receptor comprises a transgenic T cell receptor (tg-TCR) or a chimeric antigen receptor (CAR).
5. The isolated cell of claim 2 , wherein said CAR comprises:
an antigen binding domain being an antibody or an antigen-binding fragment; or
a CD3ζ; or
at least one co-stimulatory domain selected from the group consisting of CD28, CD134/OX40, CD137/4-1BB, Lck, ICOS and DAP10; or
at least two co-stimulatory domains selected from the group consisting of CD28, CD134/OX40, CD137/4-1BB, Lck, ICOS and DAP10.
6. The isolated cell of claim 5 , wherein the antigen-binding fragment is a Fab or a scFv.
7. The isolated cell of claim 1 , wherein said disease antigen is selected from the group consisting of a tumor antigen, a viral antigen, a bacterial antigen, a fungal antigen, a protozoa antigen, and a parasite antigen.
8. The isolated cell of claim 7 , wherein:
said tumor antigen is associated with a solid tumor; or
said tumor antigen is associated with a hematologic malignancy; or
said tumor antigen is selected from the group consisting of CD19, CD20, CD22, ROR1, mesothelin, CD33/IL3Ra, c-Met, PSMA, Glycolipid F77, EGFRvIII, Her2, GD2, gp100, p53, carcinoembryonic antigen (CEA), MART-1, Telomerase reverse transcriptase (TERT), Claudin-6, Receptor tyrosine-protein kinase extracellular domain (ErbB2-ECD), Receptor tyrosine-protein kinase intracellular domain (ErbB2-ICD), Histone H1.2, Histone H4, Tyrosinase, alphafetoprotein (AFP), MAGE A3, AIM-2a, AFP, ART-4, CLCA2, Cyp-B, EphA2, hTERT, iCE, FGF-5, G250, GnT-V, HST-2 (FGF-6), Livin (ML-IAP), MUC1, MUC2, PRAME, PSMA, P15, RAGE, RU1, RU2, SART-1, SART-3, SART-2, SOX10, Survivin, Survivin-2Bg, TRG, Neo-PAP, CAMEL and NY-ESO-1; or
said viral antigen is of a virus selected from the group consisting of human immunodeficiency virus (HIV), T-cell leukemia virus type 1 (TAX), influenza virus, herpes virus, papilloma virus, hepatitis viruses, ebola virus, human T-lymphotropic virus (HTLV), rubella virus, measles virus, rabies virus, lymphocytic choriomeningitis (LCM), rotavirus, mumps virus, adenovirus, BK polyomavirus (BKV), and Epstein-Barr virus (EBV).
9. The isolated cell of claim 1 , wherein said cell is further genetically modified to repress expression of at least one endogenous immunological checkpoint gene in said cell.
10. The isolated cell of claim 9 , wherein said immunological checkpoint gene is selected from the group consisting of a PD or CTLA gene.
11. The isolated cell of claim 1 , wherein said Tcm phenotype comprise a CD3 + , CD8 + , CD62L + , CD45RA − , CD45RO + signature, and optionally wherein at least 50% of the isolated cells are CD3+CD8+ cells of which at least 50% have said signature.
12. A population of cells comprising the isolated cell of claim 1 .
13. A pharmaceutical composition comprising the population of cells of claim 12 and a pharmaceutically active carrier.
14. The isolated cell of claim 1 , wherein said non-graft versus host (GVHD)-inducing anti-third party cell, being tolerance-inducing and capable of homing to the lymph nodes following transplantation, is generated by a method comprising:
(a) contacting peripheral blood mononuclear cells (PBMC) with a third party antigen or antigens in the presence of IL-21 so as to allow enrichment of antigen reactive cells; and
(b) culturing said cells resulting from step (a) in the presence of IL-21, IL-15 and IL-7 so as to allow proliferation of anti-third party cells comprising said central memory T-lymphocyte (Tcm) phenotype.
15. The isolated cell of claim 14 , wherein said method further comprises depleting CD4+ and/or CD56+ cells prior to step (a).