Compositions and methods for improved T cells
The invention provides compositions and methods for generating improved T cells having increased Ezh2 activity and methods of use thereof in the treatment of cancer and chronic infection.
1. A composition comprising an improved T-cell, wherein the T cell has increased EZH2 activity;
wherein the improved T cell has at least about a 5-fold decrease in phosphorylated EZH2 and about a 4-fold increase in H3K27me3;
wherein the T cell has been contacted with an inhibitor of a regulator of at least one of the level and activity of EZH2; and
wherein the regulator of EZH2 is selected from the group consisting of phosphatidylinositol 3-kinase (PI3K), calcineurin, JMJD3, AKT, AP-1, CDK1, CDK4/6, DNA-PK, and a combination thereof;
wherein the inhibitor is selected from the group consisting of CsA, Tacrolimus, GSK-J4, 5-Carboxy-8-hydroxyquinoline, MEK2206, SB203580, MK2206, SC79, AZD5363, LM22B-10, GDC-0068, GSK-690693, Afuersertib, AKT inhibitor VIII, A-443654, TIC10, Honokiol, Triciribine, A-674563, Prifosine, Miltefosine, SR11302, SP100030, c-JUN peptide, TanIIA, E3330, NNGH, Sulfaphenoazole, U0126 monoethanolate, IQ-1S, SM-7368, NIN-43, MEK inhibitor VII, TNAP inhibitor, FR180204, APE1 inhibitor III, (S)-AR-TURMERONE, 4-O-METHYLHONOKIOL, 5-(9-ISOPROPYL-2-MORPHOLINO-9H-PURIN-6-YL)PYRMIDIN-2-AMINE, A66, Arenobufagin, Bay80-6946, Benidipine Hydrochloride, BEX235, BKM120, BYL719, CAL-101, CH5132799, CUDC-907, GDC-0980, GSK-2126458, GSK-2334470, GSK-2636771, IPI-145, Ly-294002, PF-04691502 Dihydrate, Piceatanol, PKI-402, PP-121, PX-866, R547, Dinaciclib, BMS-265246, JNJ-7706621, AZD5438, Alvocidib, SU9516, PHA-793887, P276-00, AT7519, PHA-7677491, Milciclib (PHA-848125), SNS-032, NU6027, LDC000067, Palbociclib, Everolimus, Ly3023414, KU-57788, NU 7026, PIK-75, LTURM34, CC-115, Compound 401 and a combination thereof.
2. A composition comprising an improved T cell, wherein the T cell has increased EZH2 activity;
wherein the improved T cell has at least about a 5-fold decrease in phosphorylated EZH2 and about a 4-fold increase in H3K27me3; and,
wherein the T cell expresses an EZH2S21A protein.
3. The composition of claim 1 , wherein the inhibitor is an inhibitory RNA molecule targeting PI3K.
4. The composition of claim 1 , wherein the T cell is an anti-tumor T cell selected from the group consisting of a CART cell, a TCR-transgenic T cell, a Tumor infiltrating T cell (TIL), an autologous T cell, and an allogeneic T cell.
5. The composition of claim 1 , wherein the T cell is an anti-tumor CART cell comprising a chimeric antigen receptor (CAR) comprising at least one sequence selected from the group consisting of a binding domain, a co-stimulatory signaling domain, a cytoplasmic signaling sequence and a combination thereof.
6. The composition of claim 5 , wherein the CAR comprises at least one selected from the group consisting of a CD19 binding domain, a 4-1BB co-stimulatory signaling domain, and a CD3ζ cytoplasmic signaling sequence.
7. A composition comprising an improved T cell, wherein the T cell has increased EZH2 activity;
wherein the improved T cell has at least about a 5-fold decrease in phosphorylated EZH2 and about a 4-fold increase in H3K27me3; and
wherein the cell comprises an mRNA molecule encoding a variant of EZH2 or a mimetic of EZH2 that does not get phosphorylated by AKT.
8. The composition of claim 7 , wherein the cell is an anti-tumor T cell selected from the group consisting of a CART cell, a TCR-transgenic T cell, a TIL, an autologous T cell, and an allogeneic T cell.
9. The composition of claim 2 , wherein the T cell further expresses a modulator of at least one of the level and activity of Ezh2, wherein the modulator increases the amount of polypeptide of EZH2, increases the amount of mRNA of EZH2, increases the activity of EZH2, modulates the phosphorylation state of EZH2, decreases the level or activity of a negative regulator of EZH2, or any combination thereof.