Reducing immune tolerance induced by PD-L1
The present disclosure relates to compositions and methods for reducing immune tolerance associated with CAR T cell therapy. Embodiments of the present disclosure include isolated nucleic acid sequence comprising a nucleic acid sequence that encodes modified programmed cell death protein 1 (PD-1) and a nucleic acid sequence that encodes chimeric antigen receptor (CAR).
1. A modified cell that comprises a modified programmed cell death protein 1 (PD-1), wherein:
the modified PD-1 comprises an extracellular domain of PD-1 and a cytoplasmic domain of the PD-1 polypeptide that is truncated, or
the modified cell comprises a partial deletion of the PD-1 gene and a reduced amount of PD-1 as compared to the corresponding wild-type cell, or comprises a non-functional PD-1 gene, and
wherein the modified PD-1 comprises an extracellular domain and
(i) one of SEQ ID NOs: 12-14; or
(ii) does not comprise SEQ ID NO: 15 or 16.
2. The modified cell of claim 1 , wherein the modified cell further comprises a chimeric antigen receptor (CAR) comprising an antigen recognition domain of a specific antibody and an intracellular domain, or a modified or wild-type T cell receptor.
3. The modified cell of claim 2 , wherein the CAR binds a tumor antigen.
4. The modified cell of claim 3 , wherein the tumor antigen comprises HER2, CD19, CD20, CD22, Kappa or light chain, CD30, CD33, CD123, CD38, ROR1, ErbB3/4, EGFR, EGFRvIII, EphA2, FAP, carcinoembryonic antigen, EGP2, EGP40, mesothelin, TAG72, PSMA, NKG2D ligands, B7-H6, IL-13 receptor α 2, IL-11 receptor α, MUC1, MUC16, CA9, GD2, GD3, HMW-MAA, CD171, Lewis Y, G250/CAIX, HLA-AI MAGE A1, HLA-A1, NY-ESO-1, PSC1, folate receptor-α, CD44v7/8, 8H9, NCAM, VEGF receptors, 5T4, Fetal AchR, NKG2D ligands, CD44v6, TEM1, or TEM8.
5. The modified cell of claim 3 , wherein the tumor antigen comprises a glioma-associated antigen, carcinoembryonic antigen (CEA), β-human chorionic gonadotropin, alphafetoprotein (AFP), lectin-reactive AFP, thyroglobulin, RAGE-1, MN-CA IX, human telomerase reverse transcriptase, RU1, RU2 (AS), intestinal carboxyl esterase, mut hsp70-2, M-CSF, prostase, prostate-specific antigen (PSA), PAP, NY-ESO-1, LAGE-1a, p53, prostein, PSMA, Her2/neu, survivin and telomerase, prostate-carcinoma tumor antigen-1 (PCTA-1), MAGE, ELF2M, neutrophil elastase, ephrinB2, CD22, insulin growth factor (IGF)-I, IGF-II, IGF-I receptor, or mesothelin.
6. The modified cell of claim 2 , wherein the tumor antigen comprises CD19.
7. The modified cell of claim 2 , wherein the intracellular domain of CAR comprises a domain of 4-1BB or CD28.
8. The method of claim 1 , wherein the modified cell further comprises a TCR.
9. The modified cell of claim 1 , wherein the modified cell comprises a nucleic acid sequence that encodes the modified PD-1 comprising substitution or deletion of one or more nucleotides as compared to a nucleic acid sequence encoding an intracellular part of wild-type PD-1.
10. The modified cell of claim 1 , wherein the modified cell is a NK cell.
11. The modified cell of claim 1 , wherein the modified cell is a T cell.
12. The modified cell of claim 1 , wherein the modified cell further comprises a cytotoxic T lymphocyte-associated protein 4 (CTLA-4) receptor polypeptide, or B and T lymphocyte attenuator (BTLA) receptor polypeptide having a truncated cytoplasmic domain.
13. The modified cell of claim 1 , wherein the modified cell comprises a nucleic acid sequence that encodes the modified PD-1 comprising deletion of multiple nucleotides as compared to a nucleic acid sequence encoding an intracellular part of wild-type PD-1.
14. The modified cell of claim 1 , wherein the binding between the modified PD-1 and PD-L1 adversely affects the binding between wild-type PD-1 of the modified cell and PD-L1.
15. The modified cell of claim 2 , wherein the CAR is specific for a tumor antigen that is present on a cancer cell, and wherein the cancer cell or a cell associated with the cancer cell expresses PD-L1.
16. The modified cell of claim 2 , wherein the modified cell comprises a nucleic acid encoding the CAR and the modified PD-1.
17. A vector comprising the nucleic acid of claim 16 .
18. A pharmaceutical composition comprising a modified cell comprising the vector of claim 17 .
19. A pharmaceutical composition comprising a modified cell as claimed in claim 1 .
20. The modified cell of claim 1 , wherein the modified PD-1 comprises one of SEQ ID NOs: 12-14.
21. The modified cell of claim 1 , wherein the modified PD-1 does not comprise SEQ ID NO: 15 or 16.