Methods and compositions for cellular immunotherapy
The present disclosure relates to genetically modified T cells comprising a transgene encoding an engineered antigen specific receptor, wherein expression of an endogenous gene selected from MNK1, MNK2, or both are inhibited in the genetically modified T cell in order to enhance central memory T cell subsets in cellular immunotherapy compositions.
1. A modified T cell, comprising a chromosomal MNK gene knock out, wherein the MNK gene comprises the MNK1 gene, the MNK2 gene, or both; optionally further comprising a transgene encoding an engineered antigen specific receptor.
2. The modified T cell of claim 1 , wherein the encoded engineered antigen specific receptor is a chimeric antigen receptor (CAR), a T cell receptor (TCR), a TCR-CAR, or any combination thereof.
3. The modified T cell of claim 2 , wherein the encoded CAR comprises a binding domain comprising an scFv that specifically binds to the antigen.
4. The modified T cell of claim 2 , wherein the encoded CAR comprises at least one signaling domain.
5. The modified T cell of claim 2 , wherein the encoded T cell receptor is an enhanced affinity TCR.
6. The modified T cell of claim 2 , wherein the encoded TCR is an αβTCR or a γδTCR.
7. The modified T cell of claim 1 , wherein both MNK1 and MNK2 are knocked out.
8. The modified T cell of claim 1 , wherein the antigen is a tumor antigen, a pathogenic microorganism antigen, or an autoimmune disease antigen.
9. The modified T cell of claim 8 , wherein the tumor antigen is selected from the group consisting of human immunodeficiency virus (HIV) antigens, hepatitis C virus (HCV) antigens, hepatitis B virus (HBV) antigens, cytomegalovirus (CMV) antigens, Epstein Barr virus (EBV) antigens, parasitic antigens, ROR1, EGFR, EGFRvIII, HPV E6, HPV E7, L1-CAM, Lewis A, Lewis Y, MUC1, MUC16, PSMA, CD19, CD20, CD22, CD56, CD23, CD24, CD30, CD33, CD37, CD38, CD44v6, CD72, CD79a, CD79b, CD97, CD123, CD171, CD179a, CA125, c-MET, FcRH5, WT1, VEGF-α, VEGFR1, VEGFR2, IL-13Rα2, IL-11Rα, MAGE, MAGE-Al, ephrin A2, ephrin B2, NKG2D ligands, NY-ESO-1, TAG-72, mesothelin, glioma-associated antigen, carcinoembryonic antigen (CEA), IL-13Rα, FAP, B7H3, Kit, CA-IX, CS-1, BCMA, bcr-abl, β-human chorionic gonadotropin, α-fetoprotein (AFP), ALK, cyclin B1, lectin-reactive AFP, Fos-related antigen 1, ADRB3, thyroglobulin, RAGE-1, SSX2, AKAP-4, LCK, OY-TES1, PAX5, SART3, CLL-1, fucosyl GM1, GloboH, MN-CA IX, EPCAM, EVT6-AML, TGS5, human telomerase reverse transcriptase, PLAC1, RU1, RU2 (AS), intestinal carboxyl esterase, sLe, LY6K, M-CSF, MYCN, RhoC, TRP-2, CYP1B1, BORIS, prostase, prostate-specific antigen (PSA), PAX3, PAP, LAGE-1a, LMP2, NCAM, p53, p53 mutant, Ras mutant, gp100, prostein, OR51E2, PANX3, PSCA, hTERT, HMWMAA, HAVCR1, survivin, telomerase, legumain, sperm protein 17, SSEA-4, tyrosinase, TARP, ML-IAP, MAD-CT-1, MAD-CT-2, MelanA/MART1, XAGE1, ELF2M, ERG (TMPRSS2 ETS fusion gene), NA17, neutrophil elastase, sarcoma translocation breakpoints, NY-BR-1, androgen receptor, insulin growth factor (IGF)-I, IGF-II, IGF-I receptor, GD2, o-acetyl-GD2, GD3, GM3, GPRC5D, GPR20, CXORF61, folate receptor alpha (FRα), folate receptor beta, Tie 2, TSHR, UPK2, Tn Ag, FLT3, PRSS21, PDGFR-beta, ERBB2 (Her2/neu), CAIX, TEM1/CD248, TEM7R, CLDN6, polysialic acid, PCTA-1/Galectin 8, mut hsp70-2, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, IGLL1, and psor.
10. The modified T cell of claim 1 , further comprising an inhibited TCR gene, an inhibited HLA gene, an immunosuppression component gene, or any combination thereof.
11. The modified T cell of claim 10 , wherein the inhibited TCR gene is TRA, TRB, or both.
12. The modified T cell of claim 10 , wherein the inhibited HLA gene is a HLA class I gene or an HLA class II gene, or both.
13. The modified T cell of claim 10 , wherein the inhibited immunosuppression component gene is PD-1, CTLA-4, VISTA, BTLA, KIR, LAG3, TIM-3, A2AR, CD244 (2B4), CD160, TIGIT, PVRIG (CD112R), arginase, indoleamine 2,3-dioxygenase (IDO), IL-10, IL-4, IL-1RA, IL-35, or any combination thereof.
14. The modified T cell of claim 10 , wherein the TCR gene, the HLA gene, the immune checkpoint molecule gene, or any combination thereof is knocked out, or is inhibited at the gene level, transcriptional level, translational level or any combination thereof.
15. The modified T cell of claim 10 , wherein the inhibited TCR gene, the inhibited HLA gene, the immunosuppression component gene, or any combination thereof comprise an inhibitory nucleic acid.
16. The modified T cell of claim 15 , wherein the inhibitory nucleic acid is an antisense oligonucleotide, a double stranded RNA (dsRNA) molecule, a siRNA molecule, an endoribonuclease-prepared siRNA (esiRNA), or an shRNA molecule.
17. The modified T cell of claim 1 , wherein the T cell is a CD4+ T cell or a CD8+ T cell.
18. The modified T cell of claim 1 , wherein the T cell is a CD62L Hi T cell.
19. The modified T cell of claim 18 , wherein the modified T cell is a modified CD4+ or CD8+ T cell made from:
(a) a CD45RA Hi CD62L Hi naïve T cell-enriched CD4+ or CD8+ population, respectively;
(b) a CD45RO Hi CD62L Hi central memory T cell-enriched CD4+ or CD8+ population, respectively;
(c) a CD62L Hi naïve and central memory T cell-enriched CD4+ or CD8+ population, respectively; or
(d) a bulk CD4+ or CD8+ T cell population, respectively, in some other embodiments, a population of modified T cells is enriched for modified CD4+ or CD8+ T cells, respectively, wherein at least 50%, at least about 60%, at least about 70%, at least about 80% or at least about 90% of the modified CD4+ or CD8+ T cells, respectively, are CD62L Hi or CD62L Hi CD45RO Hi .
20. The modified T cell of claim 1 , wherein the T cell is a human T cell.