IMMUNE EFFECTOR CELL THERAPIES WITH ENHANCED EFFICACY

• US 20190000880A1
• Filed: 12/30/2016
• Published: 01/03/2019
• Est. Priority Date: 12/30/2015
• Status: Active Application

• Current Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Motz, Gregory, Mavrakias, Konstantinos John, Liu, Jinbiao, Liu, Lei, Zheng, Qiangang, Xun, Guoliang, Xiao, Qitao
• Application Number:
  US16/066,855
• Publication Number:
  US 20190000880A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 2239/28 Expressing multiple CARs, TCRs or antigens
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 38/005 Enzyme inhibitors protease inhibitors A61K38/55
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464412 CD19 or B4
• CPC Class Codes: A61K 39/464417 Receptors for tumor necrosis factors [TNF], e.g. lymphotoxin receptor [LTR], CD30
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07D 207/34 with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
• CPC Class Codes: C07D 401/14 containing three or more hetero rings
• CPC Class Codes: C07K 16/241 Tumor Necrosis Factors
• CPC Class Codes: C07K 16/2815 against CD8
• CPC Class Codes: C07K 16/2818 against CD28 or CD152
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/3092 against tumour-associated mucins
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/531 Stem-loop; Hairpin
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 5/0634 : Cells from the blood or the immune system
• CPC Class Codes: C12N 5/0636 : T lymphocytes

Abstract

Provided are the use of LSD1 inhibitors in connection with use and manufacture of immune effector cells (e.g., T cells, NK cells), e.g., engineered to express a chimeric antigen receptor (CAR), to treat a subject having a disease, e.g., a disease associated with expression of a tumor antigen.

Claims

1. A method of treating a subject, comprising administering to said subject an LSD1 inhibitor and a population of immune effector cells engineered to express a CAR.

2. The method of claim 1, wherein:

a) the LSD1 inhibitor is administered before the subject is administered said population of immune effector cells;

b) the LSD1 inhibitor is administered concurrently with said population of immune effector cells;

c) the LSD1 inhibitor is administered after the subject is administered said population of immune effector cells;

or

d) any combination of a), b) and/or c).

3. The method of claim 2, wherein the LSD1 inhibitor is administered before the subject is administered said population of immune effector cells, and wherein said administration of the LSD1 inhibitor is continued for a period of time after the administration of said population of immune effector cells.

4. The method of claim 3, wherein the administration of the LSD1 inhibitor is in an amount sufficient to increase an anti-tumor effect of said population of immune effector relative to an equivalent population of said immune effector cells administered in the absence of said LSD1 inhibitor.

7. The method of any of claim 1-4 or 6, wherein the administration or the contacting of the LSD1 inhibitor results in:

1) an increase in the proportion of naive T cells, e.g., T_SCM cells;

2) an increase in the number of naive T cells, e.g., T_SCM cells;

3) a decrease in the number of T_EM cells;

4) a decrease in the proportion of T_EM cells;

5) an increase in the proportion of CD45RA+CD62L+ T cells;

6) an increase in the number of CD45RA+CD62L+ T cells;

7) an increase in the proportion of CD45RA+CCR7+ T cells;

8) an increase in the number of CD45RA+CCR7+ T cells;

9) a decrease in the proportion of PD-1 positive immune effector cells;

10) an increase in the ratio of PD-1 negative immune effector cells/PD-1 positive immune effector cells;

11) a decrease in the proportion of PD-1+/Lag3+/Tim3+ immune effector cells;

12) an increase in the ratio of PD-1−

/Lag3−

/Tim3−

immune effector cells to PD-1+/Lag3+/Tim3+ immune effector cells;

13) an increase in the proliferation of the immune effector cells;

14) an increase in the production of cytokines (e.g., IFNg and/or IL-2) from said population of immune effector cells;

or

15) a combination of two or more of the above;

optionally as compared to cells not contacted with the LSD1 inhibitor.

19. The method or population of immune effector cells of any of the preceding claims, wherein the population of immune effector cells comprise, e.g., consist of, T cells or NK cells.

20. The method or population of immune effector cells of claim 19, wherein the population of immune effector cells comprise, e.g., consist of, T cells.

21. The method or population of immune effector cells of claim 20, wherein the T cells are CD8+ T cells, CD4+ T cells, or a combination thereof.

22. The method or population of immune effector cells of any of the preceding claims, wherein the immune effector cells are human cells.

23. The method of any of claim 1-7, 10, 12-14 or 18-22, or population of immune effector cells of any of claims 15-22, wherein the CAR comprises an antigen binding domain (which is optionally an antibody or antibody fragment, TCR or TCR fragment), a transmembrane domain, and an intracellular signaling domain (which is optionally an intracellular signaling domain comprising a costimulatory domain and/or a primary signaling domain).

24. The method or population of immune effector cells of claim 23, wherein the antigen-binding domain binds to a tumor antigen selected from a group consisting of:

TSHR, CD19, CD123, CD22, CD30, CD171, CS-1, CLL-1, CD33, EGFRvIII, GD2, GD3, BCMA, Tn Ag, PSMA, ROR1, FLT3, FAP, TAG72, CD38, CD44v6, CEA, EPCAM, B7H3, KIT, IL-13Ra2, Mesothelin, IL-11Ra, PSCA, PRSS21, VEGFR2, LewisY, CD24, PDGFR-beta, SSEA-4, CD20, Folate receptor alpha, ERBB2 (Her2/neu), MUC1, EGFR, NCAM, Prostase, PAP, ELF2M, Ephrin B2, IGF-I receptor, CAIX, LMP2, gp100, bcr-abl, tyrosinase, EphA2, Fucosyl GM1, sLe, GM3, TGS5, HMWMAA, o-acetyl-GD2, Folate receptor beta, TEM1/CD248, TEM7R, CLDN6, GPRC5D, CXORF61, CD97, CD179a, ALK, Polysialic acid, PLAC1, GloboH, NY-BR-1, UPK2, HAVCR1, ADRB3, PANX3, GPR20, LY6K, OR51E2, TARP, WT1, NY-ESO-1, LAGE-1a, MAGE-A1, legumain, HPV E6, E7, MAGE A1, ETV6-AML, sperm protein 17, XAGE1, Tie 2, MAD-CT-1, MAD-CT-2, Fos-related antigen 1, p53, p53 mutant, prostein, survivin and telomerase, PCTA-1/Galectin 8, MelanA/MART1, Ras mutant, hTERT, sarcoma translocation breakpoints, ML-IAP, ERG (TMPRSS2 ETS fusion gene), NA17, PAX3, Androgen receptor, Cyclin B1, MYCN, RhoC, TRP-2, CYP1B1, BORIS, SART3, PAX5, OY-TES1, LCK, AKAP-4, SSX2, RAGE-1, human telomerase reverse transcriptase, RU1, RU2, intestinal carboxyl esterase, mut hsp70-2, CD79a, CD79b, CD72, LAIR1, FCAR, LILRA2, CD300LF, CLEC12A, BST2, EMR2, LY75, GPC3, FCRL5, and IGLL1.

25. The method or population of immune effector cells of claim 24, wherein the tumor antigen is CD19 or BCMA.

26. The method or population of immune effector cells of claim 25, wherein:

the antigen-binding domain is an antibody or antibody fragment comprising;

(i) the amino acid sequence of a CD19 binding domain according to Tables 6-9, e.g., the amino acid sequence of CTL019 scFv domain according to Table 9 or an amino acid sequence according to SEQ ID NO;

957, or an amino acid sequence at least 95% identical thereto;

(ii) the amino acid sequence of a humanized CD19 binding domain according to Tables 6-9, e.g., the amino acid sequence of CAR2 scFv domain according to Table 9 or an amino acid sequence according to SEQ ID NO;

898, or an amino acid sequence at least 95% identical thereto;

or

(iii) the amino acid sequence of a BCMA binding domain according to Tables 11A-11B, e.g., the amino acid sequence of 139109 scFv domain according to Table 11A or an amino acid sequence according to SEQ ID NO;

967, or an amino acid sequence at least 95% identical thereto;

or

wherein the CAR comprises;

(i) the amino acid sequence of a CD19 CAR according to Tables 6-9, e.g., the amino acid sequence of CTL019 according to Table 9 or an amino acid sequence according to SEQ ID NO;

956 or an amino acid sequence at least 95% identical thereto;

(ii) the amino acid sequence of a humanized CD19 CAR according to Tables 6-9, e.g., the amino acid sequence of CAR2 according to Table 9 or an amino acid sequence according to SEQ ID NO;

902, or an amino acid sequence at least 95% identical thereto;

or

(iii) the amino acid sequence of a BCMA CAR according to Tables 11A-11B, e.g., the amino acid sequence of 139109 CAR according to Table 11A or an amino acid sequence according to SEQ ID NO;

971, or an amino acid sequence at least 95% identical thereto.

27. The method or population of immune effector cells of any of claims 23-26, wherein the transmembrane domain comprises:

(i) an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of an amino acid sequence of SEQ ID NO;

12, or a sequence with 95-99% identity to an amino acid sequence of SEQ ID NO;

12;

or

(ii) the sequence of SEQ ID NO;

12.

28. The method or population of immune effector cells of any of claim 23-27, wherein the antigen binding domain is connected to the transmembrane domain by a hinge region, wherein said hinge region comprises SEQ ID NO:

2 or SEQ ID NO;

6, or a sequence with 95-99% identity thereof.

29. The method or population of immune effector cells of any of claims 23-28, wherein the intracellular signaling domain comprises a primary signaling domain and/or a costimulatory signaling domain, wherein the primary signaling domain comprises a functional signaling domain of a protein chosen from CD3 zeta, CD3 gamma, CD3 delta, CD3 epsilon, common FcR gamma (FCER1G), FcR beta (Fc Epsilon R1b), CD79a, CD79b, Fcgamma RIIa, DAP10, or DAP12.

30. The method or population of immune effector cells of any of claims 23-29, wherein the primary signaling domain comprises:

(i) an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of an amino acid sequence of SEQ ID NO;

18 or SEQ ID NO;

20, or a sequence with 95-99% identity to an amino acid sequence of SEQ ID NO;

18 or SEQ ID NO;

20;

or

(ii) the amino acid sequence of SEQ ID NO;

18 or SEQ ID NO;

20.

31. The method or population of immune effector cells of any of claims 23-30, wherein the intracellular signaling domain comprises a costimulatory signaling domain, or a primary signaling domain and a costimulatory signaling domain, wherein the costimulatory signaling domain comprises a functional signaling domain of a protein selected from the group consisting of CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, CDS, ICAM-1, GITR, BAFFR, HVEM (LIGHTR), SLAMF7, NKp80 (KLRF1), CD160, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, NKp44, NKp30, NKp46, and NKG2D.

32. The method or population of immune effector cells of any of claims 23-31, wherein the costimulatory signaling domain comprises an amino acid sequence having at least one, two or three modifications but not more than 20, 10 or 5 modifications of an amino acid sequence of SEQ ID NO:

14 or SEQ ID NO;

16, or a sequence with 95-99% identity to an amino acid sequence of SEQ ID NO;

14 or SEQ ID NO;

16.

33. The method or population of immune effector cells of any of claims 23-32, wherein the costimulatory signaling domain comprises a sequence of SEQ ID NO:

14 or SEQ ID NO;

16.

34. The method or population of immune effector cells of any of claims 23-33, wherein the intracellular domain comprises the sequence of SEQ ID NO:

14 or SEQ ID NO;

16, and the sequence of SEQ ID NO;

18 or SEQ ID NO;

20, wherein the sequences comprising the intracellular signaling domain are expressed in the same frame and as a single polypeptide chain.

35. The method or population of immune effector cells of any of claims 23-34, wherein the CAR comprises a leader sequence comprising, e.g., consisting of, SEQ ID NO:

2.

36. The method or population of immune effector cells of any of the preceding claims, wherein the LSD1 inhibitor is:

(1) a gene editing system targeted to one or more sites of the LSD1 gene, or its corresponding regulatory elements;

(2) a nucleic acid (e.g., an siRNA or shRNA, or antisense oligonucleotide) comprising sequence complementary to a target sequence of the LSD1 gene;

(3) a protein (e.g., a dominant negative LSD1, e.g., catalytically inactive LSD1, or a dominant negative binding partner of LSD1);

(4) a small molecule;

(5) a nucleic acid encoding any of (1)-(3);

or (6) any combination of (1)-(5).

37. The method or population of immune effector cells of claim 36, wherein the LSD1 inhibitor is an shRNA or siRNA comprising sequence complementary to a target sequence of the LSD1 gene listed in Table 1, e.g., selected from SEQ ID NO:

[43] to SEQ ID NO;

[82].

38. The method or population of immune effector cells of claim 36, wherein the LSD1 inhibitor is an shRNA encoded by nucleic acid comprising any sequence encoding an anti-LSD1 shRNA of Table 1, e.g., encoded by nucleic acid comprising a sequence selected from SEQ ID NO:

[83] to SEQ ID NO;

[122].

39. The method or population of immune effector cells of claim 36, wherein the LSD1 inhibitor is nucleic acid comprising any sequence encoding an anti-LSD1 shRNA of Table 1, e.g., a sequence selected from SEQ ID NO:

[83] to SEQ ID NO;

[122].

40. The method or population of immune effector cells of claim 39, wherein said nucleic acid is disposed on a vector.

41. The method or population of immune effector cells of claim 40, wherein the vector further comprises a U6 or H1 promoter operably linked to said nucleic acid.

42. The method or population of immune effector cells of any of claims 40-41, wherein the vector is a retroviral vector, a lentiviral vector, an adenoviral vector, an adeno-associated viral (AAV) vector, a herpes simplex virus (HSV) vector, a plasmid, a minicircle, a nanoplasmid, or an RNA vector.

43. The method or population of immune effector cells of any of claims 40-42, wherein the vector further comprises sequence encoding a CAR.

44. The method or population of immune effector cells of claim 36, wherein the LSD1 inhibitor is a genome editing system specific for a sequence of the LSD1 gene (KDM1A) or its regulatory elements selected from a CRISPR genome editing system, a zinc finger nuclease genome editing system, a TALEN genome editing system and a meganuclease genome editing system.

45. The method or population of immune effector cells of claim 44, wherein the LSD1 inhibitor is a CRISPR genome editing system comprising a gRNA molecule comprising a targeting domain complementary to a sequence of the LSD1 gene (KDM1A) or its regulatory elements, e.g., comprising any one of SEQ ID NO:

[132] to [862].

46. The method or population of immune effector cells of claim 36, wherein the LSD1 inhibitor is a small molecule.

47. The method or population of immune effector cells of claim 46, wherein the small molecule is a reversible or irreversible LSD1 inhibitor.

48. The method or population of immune effector cells of claim 46 or 47, wherein the LSD1 inhibitor is:

a) GSK2699537;

b) rel-2-[[(1R,2S)-2-[4-[(4-chlorophenyl)methoxy]phenyl]cyclopropyl]amino]-1-(4-methyl-1-piperazinyl)-ethanone;

c) (R)-4-(5-(pyrrolidin-3-ylmethoxy)-2-(p-tolyl)pyridin-3-yl)benzonitrile;

d) (1S,2R)—

N-((2-methoxypyridin-3-yl)methyl)-2-phenylcyclopropan-1-amine;

e) N,N-dimethyl-1-((4-(4-(4-(piperidin-4-yl)phenyl)-1H-indazol-1-yl)phenyl)sulfonyl)piperidin-4-amine;

f) 5-(6-chloro-4′

-(methylsulfonyl)-[1,1′

-biphenyl]-3-yl)-2-(piperazin-1-yl)-1H-pyrrole-3-carbonitrile;

g) rel-N-[(1R,2S)-2-Phenylcyclopropyl]-4-Piperidinamine;

h) 2-(1R,2S)-2-(4-(Benzyloxy)phenyl)cyclopropylamino)-1-(4-methylpiperazin-1-yl)ethanone;

i) Trans-3-(3-amino-2-methylphenyl)-1-(4-hydroxycyclohexyl)-6-methyl-1H-indole-5-carbonitrile;

or

j) a pharmaceutically acceptable salt of any of the foregoing.

49. The method or population of immune effector cells of any of claims 46-48, wherein the LSD1 inhibitor is conjugated to an antibody or antigen-binding fragment thereof.

50. The method or population of immune effector cells of claim 49, wherein the antibody or antigen-binding fragment thereof recognizes an antigen on the surface of a T cell.

51. The method or population of immune effector cells of claim 50, wherein the antigen on the surface of a T cell is CD3.

52. The method or population of immune effector cells of claim 36, wherein the LSD1 inhibitor is a protein, e.g., is a dominant negative binding partner of LSD1 (e.g., a histone deacetylase (HDAC) that interacts with LSD1 or other member of the Co-REST or AR co-activator complex), or nucleic acid encoding said dominant negative binding partner of LSD1.

53. The method or population of immune effector cells of claim 36, wherein the inhibitor of LSD1 is a protein, e.g., is a dominant negative (e.g., catalytically inactive) LSD1, or nucleic acid encoding said dominant negative LSD1.

59. The method of any of claim 1-15 or 19-58, wherein the subject has a disease associated with expression of a tumor antigen, e.g., a proliferative disease, a precancerous condition, a cancer, and a non-cancer related indication associated with expression of the tumor antigen.

60. The method of claim 59, wherein the cancer is a hematologic cancer chosen from one or more of chronic lymphocytic leukemia (CLL), acute leukemias, acute lymphoid leukemia (ALL), acute myeloid leukemia (AML), B-cell acute lymphoid leukemia (B-ALL), T-cell acute lymphoid leukemia (T-ALL), chronic myelogenous leukemia (CML), B cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt'"'"'s lymphoma, diffuse large B cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell- or a large cell-follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin'"'"'s lymphoma, Hodgkin'"'"'s lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, or pre-leukemia.

61. The use or method of claim 59, wherein the cancer is selected from the group consisting of colon cancer, rectal cancer, renal-cell carcinoma, liver cancer, non-small cell carcinoma of the lung, cancer of the small intestine, cancer of the esophagus, melanoma, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin'"'"'s Disease, non-Hodgkin'"'"'s lymphoma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, solid tumors of childhood, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi'"'"'s sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers, combinations of said cancers, and metastatic lesions of said cancers.

62. The method or population of immune effector cells of any of the preceding claims, wherein the subject is a human.

63. The method or population of immune effector cells of claim 62, wherein the human has a disease associated with expression of a tumor antigen, e.g., a cancer.

67. The compound of any of claims 64-65, for use in the method of any of claim 1-14, 19-36, 46-51, or 54-63.

5. A method of increasing the therapeutic efficacy of a population of immune effector cells engineered to express a CAR, e.g., a CART19 (e.g., CTL019), comprising a step of decreasing the activity or expression of LSD1 in said cells, transiently or permanently.

6. The method of claim 5, wherein the step of decreasing the activity or expression of LSD1 in said cells comprises contacting the cells with an LSD1 inhibitor.

8. A method of treating a subject, comprising:

a) administering an LSD1 inhibitor to a subject;

b) collecting a population of immune effector cells from the subject of step a), after said administration of the LSD1 inhibitor;

c) providing said population of immune effector cells ex vivo;

d) contacting said ex vivo population of immune effector cells with the LSD1 inhibitor, wherein the contacting with the LSD1 inhibitor causes one or more of the following to occur;

1) an increase in the proportion of naive T cells, e.g., T_SCM cells;

2) an increase in the number of naive T cells, e.g., T_SCM cells;

3) a decrease in the number of T_EM cells;

4) a decrease in the proportion of T_EM cells;

5) an increase in the proportion of CD45RA+CD62L+ T cells;

6) an increase in the number of CD45RA+CD62L+ T cells;

7) an increase in the proportion of CD45RA+CCR7+ T cells;

8) an increase in the number of CD45RA+CCR7+ T cells;

9) a decrease in the proportion of PD-1 positive immune effector cells;

10) an increase in the ratio of PD-1 negative immune effector cells/PD-1 positive immune effector cells;

11) a decrease in the proportion of PD-1+/Lag3+/Tim3+ immune effector cells;

12) an increase in the ratio of PD-1−

/Lag3−

/Tim3−

immune effector cells to PD-1+/Lag3+/Tim3+ immune effector cells;

13) an increase in the proliferation of the immune effector cells;

14 an increase in the production of cytokines (e.g., IFNg and/or IL-2) from said population of immune effector cells;

or

15) a combination of two or more of the above;

as compared to a non-contacted ex vivo population of immune effector cells; and

e) administering the population of immune effector cells to a subject.

9. The method of claim 8, wherein the step of e) further comprises administering the LSD1 inhibitor to the subject of step e).

10. The method of claim 8 or 9, further comprising the step of inserting nucleic acid that encodes a CAR into cells of the ex vivo population of immune effector cells.

11. A method of making a population of immune effector cells comprising:

a) contacting a population of immune effector cells with an LSD1 inhibitor;

thereby making a population of immune effector cells,

wherein the contacting with the LSD1 inhibitor causes one or more of the following to occur;

1) an increase in the proportion of naive T cells, e.g., T_SCM cells;

2) an increase in the number of naive T cells, e.g., T_SCM cells;

3) a decrease in the number of T_EM cells;

4) a decrease in the proportion of T_EM cells;

5) an increase in the proportion of CD45RA+CD62L+ T cells;

6) an increase in the number of CD45RA+CD62L+ T cells;

7) an increase in the proportion of CD45RA+CCR7+ T cells;

8) an increase in the number of CD45RA+CCR7+ T cells;

9) a decrease in the proportion of PD-1 positive immune effector cells;

10) an increase in the ratio of PD-1 negative immune effector cells/PD-1 positive immune effector cells;

11) a decrease in the proportion of PD-1+/Lag3+/Tim3+ immune effector cells;

12) an increase in the ratio of PD-1−

/Lag3−

/Tim3−

immune effector cells to PD-1+/Lag3+/Tim3+ immune effector cells;

13) an increase in the proliferation of the immune effector cells;

14 an increase in the production of cytokines (e.g., IFNg and/or IL-2) from said population of immune effector cells;

or

15) a combination of two or more of the above;

as compared to a non-contacted population of immune effector cells.

12. The method of claim 11, further comprising the step of b) inserting nucleic acid that encodes the CAR into cells of the population of immune effector cells.

13. The method of claim 12 wherein said contacting of step a) occursprior to;

2) concurrently with;

3) after;

or

4) any combination of two or more of

     1) to

     3) above;

said inserting of step b).

14. The method of any of claims 11-13, wherein the contacting of step a), and optionally the inserting of step b), is ex vivo.

15. A population of immune effector cells, made by the method of any of claims 11-14.

54. A method of treating a subject in need thereof, comprising administering to said subject an effective amount of the population of immune effector cells of any of claims 15-18.

55. The method of claim 54, wherein the method further comprises administering to said subject an LSD1 inhibitor.

56. The method of claim 55, wherein the subject receives a pre-treatment of the LSD1 inhibitor, prior to the administration of the population of immune effector cells.

57. The method of any of claims 55-56, wherein the subject receives concurrent treatment with an LSD1 inhibitor and the population of immune effector cells.

58. The method of any of claims 55-57, wherein the subject receives treatment with an LSD1 inhibitor after administration of the population of immune effector cells.

16. A population of immune effector cells engineered to express a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen-binding domain, a transmembrane domain, and an intracellular signaling domain, and wherein expression and/or function of LSD1 in said cell has been reduced or eliminated.

17. The population of immune effector cells of claim 16, comprising an LSD1 inhibitor.

18. The population of immune effector cells of claim 16, wherein the population of immune effector cells has been contacted with an LSD1 inhibitor.

64. A compound selected from N,N-dimethyl-1-((4-(4-(4-(piperidin-4-yl)phenyl)-1H-indazol-1-yl)phenyl)sulfonyl)piperidin-4-amine and 5-(6-chloro-4′

-(methylsulfonyl)biphenyl-3-yl)-2-(piperazin-1-yl)-1H-pyrrole-3-carbonitrile;

or a pharmaceutically acceptable salt thereof.

65. A pharmaceutically acceptable salt of the compound of claim 64.

66. The compound of any of claims 64-65, for use in therapy.

68. A composition for use in ex vivo manufacturing a population of immune effector cells, comprising an LSD1 inhibitor.

69. The composition of claim 68, wherein the LSD1 inhibitor is a small molecule.

70. The composition of claim 69, wherein the concentration of the small molecule ranges from about 0.001 nM to about 10 mM, e.g., from about 0.001 nM to about 100 nM.

71. The composition of claim 70, wherein the concentration of the small molecule ranges from about 0.1 uM to about 10 uM.

72. The composition of any of claims 68-71, wherein the population of immune effector cells comprises cells engineered to express a CAR.

73. An LSD1 inhibitor, for use in treating a subject, wherein said subject has received, is receiving, or is about to receive therapy comprising an immune effector cell, e.g., an immune effector cell engineered to express a CAR.

74. An LSD1 inhibitor, for use in the manufacture of an immune effector cell, e.g., an immune effector cell engineered to express a CAR.

75. A method of manufacturing an immune effector cells comprising introducing into said cells nucleic acid encoding a CAR, wherein the nucleic acid integrates into the genome of said cell within the LSD1 gene, such that LSD1 expression and/or function is reduced.