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Msg  4160 of 4338  at  9/17/2020 1:19:35 AM  by

JBWIN


Building IP: Juno Therapeutics/Fred Hutchinson Cancer Research Center re "COMBINATION OF A CELL THERAPY AND A GAMMA SECRETASE INHIBITOR"

 
United States Patent Application20200289565
Kind CodeA1
GREEN; Damian J. ; et al.September 17, 2020

COMBINATION OF A CELL THERAPY AND A GAMMA SECRETASE INHIBITOR

Abstract

Provided herein are combination therapies involving administration of an immunotherapy involving a cell therapy, such as a T cell therapy, and an inhibitor of gamma secretase. Also provided are methods for engineering, preparing, and producing the cells, compositions containing the cells and/or gamma secretase inhibitor, and kits and devices containing and for using, producing and administering the cells and/or gamma secretase inhibitor, such as in accord with the provided combination therapy methods.


Inventors:GREEN; Damian J.; (Seattle, WA) ; RIDDELL; Stanley R.; (Seattle, WA) ; WORKS; Melissa; (Seattle, WA)
Applicant:
NameCityStateCountryType

Juno Therapeutics, Inc.
Fred Hutchinson Cancer Research Center

Seattle
Seattle

WA
WA

US
US
Assignee:Juno Therapeutics, Inc.
Seattle
WA

Fred Hutchinson Cancer Research Center
Seattle
WA

Family ID:1000004900695
Appl. No.:16/761770
Filed:November 6, 2018
PCT Filed:November 6, 2018
PCT NO:PCT/US2018/059510
371 Date:May 5, 2020

Related U.S. Patent Documents

Application NumberFiling DatePatent Number
62665450May 1, 2018
62582937Nov 7, 2017
62582308Nov 6, 2017

Current U.S. Class:1/1
Current CPC Class:C07K 14/7151 20130101; A61K 31/55 20130101; A61K 35/17 20130101; A61P 35/00 20180101
International Class:A61K 35/17 20060101 A61K035/17; A61K 31/55 20060101 A61K031/55; A61P 35/00 20060101 A61P035/00; C07K 14/715 20060101 C07K014/715

Claims



1. A method of treatment, the method comprising: (a) administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of immune cells expressing a recombinant receptor; and (b) administering to the subject a compound of the structure: ##STR00022## or a stereoisomer thereof, or a pharmaceutically acceptable salt or hydrate of either of the foregoing.

2. A method of treatment, the method comprising administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of immune cells expressing a recombinant receptor, wherein, at the time of initiation of the administration of the cell therapy, the subject has been previously administered, and/or is undergoing treatment with, a compound having the structure: ##STR00023## or a stereoisomer thereof, or a pharmaceutically acceptable salt or hydrate of either of the foregoing.

3. A method of treatment, the method comprising administering to a subject having a disease or disorder a compound having the structure: ##STR00024## or a stereoisomer thereof, or a pharmaceutically acceptable salt or hydrate of either of the foregoing, wherein, at the time of initiation of the administration, the subject has been previously administered, and/or is undergoing treatment with, a cell therapy, said cell therapy comprising a dose of immune cells expressing a recombinant receptor.

4. The method of claim 1 or claim 3, wherein the initiation of administration of the compound is prior to, concurrently with or subsequently to initiation of administration of the cell therapy.

5. The method of claim 4, wherein the compound is administered prior to initiation of administration of the cell therapy.

6. The method of claim 1 or claim 2, wherein initiation of administration of the compound is within, or within about, 1 hours, 2 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours or 1 week prior to the initiation of the administration of the cell therapy.

7. The method of claim 6, wherein the compound is administered subsequently to initiation of administration of the cell therapy.

8. A method of treatment, the method comprising (a) administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of immune cells expressing a recombinant receptor; and (b) subsequently to the administration in (a), administering to the subject a compound having the structure: ##STR00025## or a stereoisomer thereof, or a pharmaceutically acceptable salt or hydrate of either of the foregoing.

9. A method of treatment, the method comprising administering a compound to a subject having a disease or disorder, wherein, at the time of initiation of the administration, the subject has been previously administered, and/or is undergoing treatment with, a cell therapy, said cell therapy comprising a dose of immune cells a expressing recombinant receptor, wherein the compound is a compound having the structure: ##STR00026## or a stereoisomer thereof, or a pharmaceutically acceptable salt or hydrate of any of the foregoing.

10. The method of any of claims 1-9, wherein the cell therapy or the recombinant receptor specifically binds to a target antigen associated with the disease or disorder.

11. The method of any of claims 1-10, wherein the cell therapy or the recombinant receptor specifically binds to a target antigen, wherein the compound is capable of inhibiting cleavage of the target antigen and/or wherein the target antigen is cleaved by a gamma secretase.

12. The method of any of claims 1 to 11, wherein the cell therapy and/or the recombinant receptor specifically binds to or targets a BCMA.

13. The method of claim 12, wherein the cell therapy and/or the recombinant receptor binds to BCMA expressed on the surface of plasma cells and/or on the surface of multiple myeloma cells.

14. The method of any of claims 1 to 11, wherein the cell therapy and/or the recombinant receptor specifically binds to a Muc 1.

15. The method of any one of claims 1-14, wherein the recombinant receptor is a chimeric antigen receptor.

16. The method of any one of claims 1-15, wherein the compound is capable of inhibiting, or inhibiting an activity or function of, a gamma secretase.

17. The method of any one of claims 1-16, wherein the compound, stereioisomer, pharmaceutically acceptable sale or hydrate inhibits or is capable of inhibiting intramembrane cleavage of BCMA.

18. The method of any of claims 1-17, wherein the subject comprises plasma cells, or cancer cells or myeloma cells or cells expressing plasma cell markers, expressing surface BCMA.

19. The method of claim 17 or claim 18, wherein: the subject has a cancer in which cells of the cancer in the subject (i) express CD138, surface CD38 or a surface plasma cell marker or are derived from plasma cells and (ii) comprise low expression of surface B cell maturation antigen (BCMA) and/or a level of expression of surface BCMA below a threshold level; and/or the method further comprises selecting a subject that has a cancer in which cells of the cancer in the subject (i) express CD138, surface CD38 or a surface plasma cell marker or are derived from plasma cells and (ii) comprise low expression of surface B cell maturation antigen (BCMA) and/or a level of expression of surface BCMA below a threshold level.

20. The method of any one of claims 1-19, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered orally, subcutaneously or intravenously.

21. The method of any of claims 1-20, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered orally.

22. The method of any one of claims 1-21, wherein: the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered at least or is administered six times daily, five times daily, four times daily, three times daily, twice daily, once daily, every other day, three times a week, at least once a week, twice a week or only one time; or the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered six times daily, five times daily, four times daily, three times daily, twice daily, once daily, every other day, three times a week, twice a week, once a week, or only one time for the duration of the compound treatment period.

23. The method of any of claims 1-22, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered once every other day, optionally for the duration of the compound treatment period.

24. The method of any of claims 1-22, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered no more than once per day, optionally for the duration of the compound treatment period.

25. The method of any of claims 1-22, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered once three times per week, optionally for the duration of the compound treatment period.

26. The method of any one of claims 22-25, wherein the administration is for the duration of the compound treatment period.

27. The method of any of claims 22-26, wherein the compound treatment period: is at least or at least about, or is or is about, 14 days; is at least or at least about, or is or is about, 21 days; or is at least or at least about, or is or is about, 28 days.

28. The method of any of claims 1-27, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, comprises administration, at an amount of 1.0 mg to 100 mg, 1.0 mg to 50 mg, 1.0 mg to 25 mg, 1.0 mg to 10 mg, 1.0 mg to 5.0 mg, 5.0 mg to 100 mg, 5.0 mg to 50 mg, 5.0 mg to 25 mg, 5.0 mg to 10 mg, 10 mg to 100 mg, 10 mg to 50 mg, 10 mg to 25 mg, 25 mg to 100 mg, 25 mg to 50 mg.

29. The method of any of claims 1-28, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, is at an amount that is at least or at least about or is or is about 0.5 mg, 1.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 25 mg, 50 mg, 100 mg, 250 mg or 500 mg.

30. The method of any of claims 1-29, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, is at an amount that is at least or at least about or is or is about 5.0 mg, 10.0 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 50 mg, or 100 mg.

31. The method of any of claims 1-30, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, is at an amount that is at least or at least about or is or is about 50 mg.

32. The method of any of claims 1-31, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, at an amount of at or about 50 mg.

33. The method of any of claims 1-30, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, is at an amount that is at least or at least about or is or is about 25 mg.

34. The method of any of claims 1-30 and 33, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, at an amount of at or about 25 mg.

35. The method of any of claims 1-30, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered, or one or more or each administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate during the compound treatment period, independently, at an amount of at or about 0.1 mg/kg weight of the subject, 0.2 mg/kg weight of the subject, 0.21 mg/kg weight of the subject, 0.22 mg/kg weight of the subject, 0.23 mg/kg weight of the subject, 0.24 mg/kg weight of the subject, 0.25 mg/kg weight of the subject, 0.3 mg/kg weight of the subject, 0.4 mg/kg weight of the subject, or 0.5 mg/kg weight of the subject.

36. The method of any of claims 1 to 35, wherein the cell therapy and/or recombinant receptor specifically binds to or targets a target antigen associated with the disease or disorder and the compound inhibits cleavage of the target antigen and/or inhibits shedding of the target antigen.

37. The method of any one of claims 1 to 36, wherein administration of the , stereoisomer, pharmaceutically acceptable salt or hydrate: decreases BCMA cleavage or shedding from cells, optionally plasma cells, in the subject, by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of BCMA cleavage or shedding from cells in the subject prior to administration of the compound; decreases a level or amount of BCMA detected in the serum of a subject by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level or amount of BCMA in the serum of the subject prior to administration of the compound; and/or increases expression of surface BCMA on cells, optionally plasma cells by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of surface BCMA on the cells in the subject prior to administration of the compound.

38. The method of any one of claims 1 to 36, wherein said administration of the , compound, stereoisomer, pharmaceutically acceptable salt or hydrate: results in a decrease, at a time point subsequent to said administration, in BCMA cleavage or shedding from cells, optionally plasma cells, optionally multiple myeloma cells, in the subject, said decrease being at or greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, compared to the level of BCMA cleavage or shedding from cells in the subject prior to administration of the compound, optionally wherein said time point is at or about 24, 48, 36, or 72 hours, or 1 week subsequent to initiation of said administration; results in a decrease, at a time point subsequent to said administration, in a level or amount of BCMA detected or measured in the serum of the subject by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level or amount of BCMA in the serum of the subject prior to administration of the compound, optionally wherein said time point is at or about 24, 48, 36, or 72 hours, or 1 week subsequent to initiation of said administration; and/or results in an increase, at a time point subsequent to said administration, of expression of surface BCMA on cells, optionally plasma cells, optionally multiple myeloma cells, in the subject, by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of surface BCMA on the cells in the subject prior to administration of the compound, optionally wherein said time point is at or about 24, 48, 36, or 72 hours, or 1 week subsequent to initiation of said administration.

39. The method of any of claims 1-38, wherein administration of the compound: decreases cleavage or shedding of a target antigen specifically bound by the cell therapy and/or recombinant receptor, optionally BCMA or Muc1, from cells, optionally plasma cells, by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of cleavage or shedding of the target or target antigen from cells in the subject prior to administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate; decreases the level or amount of a target antigen specifically bound by the cell therapy and/or recombinant receptor, optionally BCMA or Muc1, detected in the serum of a subject by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level or amount of the target or target antigen in the serum of the subject prior to administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate; and/or increases surface expression of a target antigen specifically bound by the cell therapy and/or recombinant receptor, optionally BCMA or Muc1, on cells, optionally plasma cells, by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of surface target or target antigen on the cells in the subject prior to administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate.

40. The method of any of claims 1-38, wherein administration of the compound: results in a decrease, at a time point subsequent to said administration, in cleavage or shedding of a target antigen specifically bound by the cell therapy and/or recombinant receptor, optionally BCMA or Muc1, from cells, optionally plasma cells, said decrease being at or greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of cleavage or shedding of the target or target antigen from cells in the subject prior to administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate; results in a decrease, at a time point subsequent to said administration, in the level or amount of a target antigen specifically bound by the cell therapy and/or recombinant receptor, optionally BCMA or Muc1, detected in the serum of a subject, said decrease being at or greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level or amount of the target or target antigen in the serum of the subject prior to administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate; and/or results in an increase, at a time point subsequent to said administration, in the surface expression of a target antigen specifically bound by the cell therapy and/or recombinant receptor, optionally BCMA or Muc1, on cells, optionally plasma cells, the increase being at or greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of surface target or target antigen on the cells in the subject prior to administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate.

41. The method of any of claims 1-40, wherein the administration comprises administration of a compound of the structure: ##STR00027##

42. The method of any of claims 1-41, wherein the disease or disorder is a cancer.

43. The method of claim 42, wherein the cancer is a B cell malignancy.

44. The method of claim 42 or claim 43, wherein the cancer is multiple myeloma, plasmacytoma, a cancer of plasma cell origin and/or a cancer of B cell origin.

45. The method of claim 42 or claim 44, wherein the cancer is a multiple myeloma.

46. The method of any of claims 42-45, wherein the cancer is a relapsed or refractory cancer, optionally a relapsed or treatment refractory multiple myeloma.

47. The method of any of claims 1-46, wherein the method further comprises, prior to said administration of said cell therapy, selecting a subject for said administration having a cancer that, or the cells of which, (i) express CD138, surface CD38 or a surface plasma cell marker or are derived from plasma cells and, optionally, (ii) comprise low expression of surface B cell maturation antigen (BCMA) and/or a level of expression of surface BCMA below a threshold level.

48. The method of claim 47, wherein the subject administered said cell therapy and/or said compound is the selected subject.

49. The method of claim 47 or claim 48, wherein the threshold level of expression of surface BCMA is lower than the average or median expression or level of surface BCMA on plasma cells in a plurality of control subjects, optionally wherein the plurality of control subjects is a group of healthy or normal subjects.

50. The method of any of claims 47-49, wherein; the low expression of surface BCMA is present when less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma marker or phenotype or the cancer cells, in the subject express surface BCMA; the threshold level of surface BCMA is less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma marker or phenotype or the cancer cells, in the subject that express surface BCMA.

51. The method of any of claims 47-50, wherein expression of surface BCMA is determined by flow cytometry and/or an immunoassay.

52. The method of any of claims 47-51, wherein (a) the ability of the antigen binding domain of the recombinant receptor, optionally chimeric antigen receptor, to bind to BCMA expressed on the surface of a target cell, or (b) a measure indicative of function or activity of the recombinant receptor, optionally the chimeric antigen receptor, to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a concentration or amount of a soluble or shed form of the BCMA.

53. The method of claim 52, wherein the concentration or amount is a concentration or amount of the soluble or shed BCMA capable of blocking or reducing or substantially blocking or reducing binding or a measure of function or activity associated with a reference anti-BCMA recombinant receptor or a reference anti-BCMA binding domain, under the same or substantially the same conditions, or is a concentration or amount present in a biological sample.

54. The method of claim 52, wherein the concentration or amount of the soluble or shed form of the BCMA is a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder.

55. The method of any of claims 15-54, wherein the chimeric antigen receptor (CAR) comprises an extracellular antigen-recognition domain that specifically binds to the antigen and an intracellular signaling domain comprising an ITAM.

56. The method of any of claims 1-54, wherein the recombinant receptor is a chimeric antigen receptor comprising an antigen-binding domain comprising: a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 173, 174 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 176, 177 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 178, 179 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 180, 181 and 182, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 186, 187 and 185, respectively; or a V.sub.H region the sequence set forth in SEQ ID NO: 24 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:24; and contains a V.sub.L region comprising the sequence set forth in SEQ ID NO:25 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:25; or an scFv comprising the sequence of amino acids set forth in SEQ ID NO:188 or a sequence of amino acids at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:188.

57. The method of claim 56, wherein antigen-binding domain or the chimeric antigen receptor specifically binds BCMA.

58. The method of any of claims 55-57, wherein the intracellular signaling domain comprises and intracellular domain of a CD3-zeta (CD3.zeta.) chain.

59. The method of any of claims 55-58, wherein the chimeric antigen receptor (CAR) further comprises a costimulatory signaling region.

60. The method of claim 59, wherein the costimulatory signaling region comprises a signaling domain derived from CD28 or 4-1BB, optionally human CD28 or human 4-1BB.

61. The method of claim 59 or claim 60, wherein the costimulatory signaling region is a domain derived from 4-1BB, optionally human 4-1BB.

62. The method of any one of claims 1-61, wherein the subject is a human.

63. The method of any of claims 1-62, wherein the BCMA is human BCMA or the target antigen is a human antigen.

64. The method of any of claims 1-63, wherein the immune cells comprise T cells or NK cells.

65. The method of claim 64, wherein the cell therapy is a T cell therapy and the dose of immune cells comprises T cells.

66. The method of claim 64 or claim 65, wherein the T cells are CD4+ and/or CD8+.

67. The method of any of the claims 64-66, wherein the T cells are primary T cells obtained from a subject.

68. The method of any of claims 1-67, wherein the cell therapy comprises cells that are autologous to the subject.

69. The method of any of claims 1-68, wherein the cell therapy comprises cells that are allogeneic to the subject.

70. The method of any of claims 1-69, wherein the administration of the cell therapy comprises the administration of from or from about 1.times.10.sup.5 to 5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), the cell therapy comprises the administration of from or from about 1.times.10.sup.5 to 1.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), from or from about 5.times.10.sup.5 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs) or from or from about 1.times.10.sup.6 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), each inclusive.

71. The method of any of claims 1-70, wherein the administration of the cell therapy comprises the administration of no more than 5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), no more than 2.5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), no more than 1.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), no more than 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), no more than 0.5.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), no more than 1.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), no more than 0.5.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs).

72. The method of any of claims 1-69, wherein the administration of the cell therapy comprises between at or about 2.5.times.10.sup.7 CAR-expressing T cells and 1.2.times.10.sup.9 CAR-expressing T cells, between at or about 5.0.times.10.sup.7 CAR-expressing T cells and 4.5.times.10.sup.8 CAR-expressing T cells, between at or about 1.5.times.10.sup.8 CAR-expressing T cells and 3.0.times.10.sup.8 CAR-expressing T cells, each inclusive.

73. The method of any of claims 1-69, wherein the cell therapy comprises between at or about 50.times.10.sup.6 and 450.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), between at or about 150.times.10.sup.6 and 450.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), between at or about 250.times.10.sup.6 and 450.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), between at or about 350.times.10.sup.6 and 450.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs), each inclusive.

74. The method of 1 and 2-73, wherein initiation of administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is within, or within about, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 14 days, 21 days, 28 days or more after the initiation of the administration of the cell therapy.

75. The method of any of claims 1 and 2-74, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered at a time in which: the number of cells of the cell therapy detectable in the blood from the subject is decreased compared to in the subject at a preceding time point after initiation of the administration of the cells; the number of cells of the cell therapy detectable in the blood is less than or less than about 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 50-fold or 100-fold or less the peak or maximum number of the cells of the cell therapy detectable in the blood of the subject after initiation of administration of the administration of the cells; and/or at a time after a peak or maximum level of the cells of the cell therapy are detectable in the blood of the subject, the number of cells of or derived from the cells detectable in the blood from the subject is less than less than 10%, less than 5%, less than 1% or less than 0.1% of total peripheral blood mononuclear cells (PBMCs) in the blood of the subject.

76. The method of any of claims 1 and 3-75, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered for a compound treatment period of up to 7 days, 14 days, 21 days, 28 days or more after initiation of the administration of the cells.

77. The method of claim 27 or claim 76, wherein the stereoisomer, pharmaceutically acceptable salt or hydrate is administered every three days for the compound treatment period.

78. The method of claim 27 or claim 76, wherein the stereoisomer, pharmaceutically acceptable salt or hydrate is administered three times a week for the compound treatment period.

79. The method of any of any of claims 1 and 3-75, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is administered on days 2, 4, 7, 9, 11, 14, 16, and 18 after initiation of the administration of the cells.

80. The method of any of any of claims 1 and 3-79, wherein at the time of or just prior to initiation of administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate the disease or condition has relapsed in the subject.

81. The method of any of claims 1-80, wherein the method further comprises administering a lymphodepleting chemotherapy prior to administration of the cell therapy and/or wherein the subject has received a lymphodepleting chemotherapy prior to administration of the cells.

82. The method of claim 81, wherein the lymphodepleting chemotherapy comprises administering fludarabine and/or cyclophosphamide to the subject.

83. The method of any of claims 1-82, wherein the method further comprises administering a steroid, optionally wherein the steroid is administered prior to, concurrently with and/or subsequently to initiation of administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate, optionally wherein the steroid is administered during the compound treatment period.

84. The method of claim 83, wherein the steroid is or comprises dexamethasone.

85. The method of any one of claims 1-84, wherein the cell therapy exhibits increased or prolonged expansion and/or persistence in the subject as compared to a method in which the cell therapy is administered to the subject in the absence of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate.

86. The method of any one of claims 1-85, wherein the method thereby prevents, reduces or ameliorates one or more symptoms or outcomes of the disease or disorder.

87. A combination, comprising: (a) immune cells expressing a recombinant receptor; and (b) a compound having the structure: ##STR00028## or a stereoisomer thereof, or a pharmaceutically acceptable salt or hydrate of any of the foregoing.

88. The combination of claim 87, wherein the compound inhibits or reduces or is capable of inhibiting or reducing intramembrane cleavage of BCMA.

89. The combination of claim 87 or claim 88, wherein the recombinant receptor and/or cell therapy targets or specifically binds to a target antigen associated with the disease or disorder.

90. The combination of any of claims 87-89, wherein the recombinant antigen receptor or cell therapy targets or specifically binds to a BCMA.

91. The combination of claim 90, wherein the cell therapy or recombinant receptor specifically bind to BCMA expressed on the surface of plasma cells, optionally multiple myeloma cells.

92. The combination of claim 91, wherein binding of the recombinant antigen receptor to surface BCMA or a measure indicative of function or activity of the recombinant receptor-expressing cells, optionally CAR-expressing cells, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA recombinant receptor, optionally, anti-BCMA CAR, in the same assay.

93. The combination of any of claims 87-92, wherein the recombinant receptor is a chimeric receptor, which optionally is a chimeric antigen receptor (CAR).

94. The combination of claim 93, wherein the recombinant receptor is a chimeric antigen receptor (CAR) and the chimeric antigen receptor comprises an extracellular antigen-recognition domain that specifically binds to the antigen and an intracellular signaling domain comprising an ITAM.

95. The combination of any of claims 87-94, wherein the recombinant receptor is a chimeric antigen receptor comprising an antigen-binding domain comprising: a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 173, 174 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 176, 177 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 178, 179 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 180, 181 and 182, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 186, 187 and 185, respectively; or a V.sub.H region the sequence set forth in SEQ ID NO: 24 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:24; and contains a V.sub.L region comprising the sequence set forth in SEQ ID NO:25 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:25; or an scFv comprising the sequence of amino acids set forth in SEQ ID NO:188 or a sequence of amino acids at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:188.

96. The combination of claim 95, wherein antigen-binding domain or the chimeric antigen receptor specifically binds BCMA.

97. The combination of any of claims 94-96, wherein the intracellular signaling domain comprises and intracellular domain of a CD3-zeta (CD3.zeta.) chain.

98. The combination of any of claims 94-97, wherein the chimeric antigen receptor (CAR) further comprises a costimulatory signaling region.

99. The combination of claim 98, wherein the costimulatory signaling region comprises a signaling domain derived from CD28 or 4-1BB, optionally human CD28 or human 4-1BB.

100. The combination of claim 98 or claim 99, wherein the costimulatory signaling region is a domain derived from 4-1BB, optionally human 4-1BB.

101. The combination of any of claims 88-100, wherein the BCMA is human BCMA or the target antigen is a human antigen.

102. The combination of any of claims 87-101, wherein the immune cells comprise T cells or NK cells.

103. The combination of claim 102, wherein the immune cells comprise T cells.

104. The combination of claim 102 or claim 103, wherein the T cells are CD4+ and/or CD8+.

105. The combination of any of claims 1102-104, wherein the T cells are primary T cells obtained from a subject.

106. The combination of any of claims 87-105, wherein the immune cells are formulated as a pharmaceutical composition for administration to a subject, optionally wherein the cells are formulated for administration in one or more unit doses for treating a disease or condition.

107. The combination of any of claims 87-106, wherein the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is formulated as a pharmaceutical composition for administration to a subject, optionally wherein the compound is formulated for administration in one or more unit doses.

108. The combination of any of claims 87-107, comprising the compound of the structure: ##STR00029##

109. A kit comprising the combination of any of claims 87-108 and instructions for administering the immune cells, and/or for administering the compound, stereoisomer, pharmaceutically acceptable salt or hydrate to a subject having a disease or disorder.

110. The kit of claim 109, wherein the instructions specify the administering of the immune cells and/or the administering of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is according to the methods of any of claims 1-86.

111. The kit of claim 109 or claim 110, further comprising a reagent for detecting expression of B cell maturation antigen (BCMA) on the surface of a cell, and instructions for administering the compound, stereoisomer, pharmaceutically acceptable salt or hydrate to a subject based on the results of use of the reagent for detecting BCMA on the surface of cells of a cancer in the subject, optionally wherein the cells of the cancer express CD138, surface CD38 or a surface plasma cell marker or are derived from plasma cells.

112. The kit of claim 111, wherein the instructions specify administering the compound, stereoisomer, pharmaceutically acceptable salt or hydrate to the subject if the cells comprise low expression of surface BCMA and/or a level of expression of surface BCMA below a threshold level.

113. The kit of claim 112, wherein the threshold level of expression of surface BCMA is lower than the average or median expression or level of surface BCMA on plasma cells in a plurality of control subjects, optionally wherein the control subjects are a group of healthy or normal subjects.

114. The kit of claim 112 or claim 113, wherein; the low expression of surface BCMA is present when less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma marker or phenotype or the cancer cells, in the subject express surface BCMA; or the threshold level of surface BCMA is less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma marker or phenotype or the cancer cells, in the subject that express surface BCMA.

115. The kit of any one of claims 109-114, wherein the instructions specify administering the compound, stereoisomer, pharmaceutically acceptable salt or hydrate or one or more unit doses thereof, to a subject having a disease or disorder prior to, concurrently with or after initiation of administration of a dose of the immune cells to the subject.

116. The kit of claim 115, wherein the instructions specify administering the compound, stereoisomer, pharmaceutically acceptable salt or hydrate, or one or more unit doses thereof, to a subject having a disease or disorder prior to initiation of administration of a dose of the immune cells to the subject.

117. The kit of claim 116, wherein the instructions specify administering the compound, stereoisomer, pharmaceutically acceptable salt or hydrate within, or within about, 1 hours, 2 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours or 1 week prior to the initiation of the administration of the cell therapy.

118. The kit of claim 115, wherein the instructions specify administering the compound, stereoisomer, pharmaceutically acceptable salt or hydrate, or one or more unit doses thereof, to a subject having a disease or disorder after initiation of administration of a dose of the immune cells to the subject.

119. A kit, comprising: (a) a compound having the structure: ##STR00030## or a stereoisomer thereof, or a pharmaceutically acceptable salt or hydrate of either of the foregoing, optionally wherein the compound is formulated in one or more unit doses; and (b) instructions for administering the compound to a subject after initiation of administration of a cell therapy to a subject, the cell therapy comprising a dose of immune cells expressing a recombinant receptor.

120. The kit of claim 119, wherein the instructions specify the administering of the immune cells and/or the administering of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is according to the methods of any of claims 1-86.

121. The kit of any one of claims 109 to 115, and 118 to 120, wherein the instructions specify initiation of administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate is within, or within about, 1 hours, 2 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 14 days, 21 days, 28 days or more after the initiation of the administration of the dose of immune cells.

122. The kit of any of any one of claims 109 to 115 and 118 to 121, wherein the instructions specify the administration of or initiation of the administration of the compound at a time in which: the number of cells of the cell therapy detectable in the blood from the subject is decreased compared to in the subject at a preceding time point after initiation of the administration of the cells, optionally wherein the preceding time is within 1, 2, 3, or 4 weeks of initiation of said cell therapy; the number of cells of the cell therapy detectable in the blood is less than or less than about 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 50-fold or 100-fold or less the peak or maximum number of the cells of the cell therapy detectable in the blood of the subject after initiation of administration of the administration of the cells; and/or at a time after a peak or maximum level of the cells of the cell therapy are detectable in the blood of the subject, the number of cells of or derived from the cells detectable in the blood from the subject is less than less than 10%, less than 5%, less than 1% or less than 0.1% of total peripheral blood mononuclear cells (PBMCs) in the blood of the subject.

123. The kit of any of any of claims 109 to 115 and 118 to 122, wherein the instructions specify the compound is for administration at a time at which the disease or condition has relapsed in the subject following treatment with the cell therapy, optionally following an objective response, and/or the subject is determined to have progressed following treatment with the cell therapy and/or a reduction or loss of target antigen has been observed in the subject following treatment with the cell therapy.

124. The kit of any of claims 119-123, wherein the recombinant antigen receptor specifically binds to a target antigen associated with the disease or disorder.

125. The kit of claim 124, wherein the target antigen is Muc1, optionally human Muc1.

126. The kit of claim 124, wherein the target antigen is BCMA, optionally human BCMA.

127. The kit of claim 126, wherein binding of the recombinant receptor to surface BCMA or a measure indicative of function or activity of the recombinant receptor-expressing cells, optionally CAR-expressing cells, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA recombinant receptor, optionally, anti-BCMA CAR, in the same assay.

128. The kit of any of claims 109-127, wherein the recombinant receptor is a chimeric receptor, which optionally is a chimeric antigen receptor (CAR).

129. The kit of claim 128, wherein the chimeric antigen receptor (CAR) comprises an extracellular antigen-recognition domain that specifically binds to the antigen and an intracellular signaling domain comprising an ITAM.

130. The kit of any of claims 109-129, wherein the recombinant receptor is a chimeric antigen receptor comprising an antigen-binding domain comprising: a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 173, 174 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 176, 177 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 178, 179 and 175, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively; a V.sub.H region comprising a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 180, 181 and 182, respectively and a V.sub.L region comprising a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 186, 187 and 185, respectively; or a V.sub.H region the sequence set forth in SEQ ID NO: 24 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:24; and contains a V.sub.L region comprising the sequence set forth in SEQ ID NO:25 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:25; or an scFv comprising the sequence of amino acids set forth in SEQ ID NO:188 or a sequence of amino acids at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:188.

131. The combination of claim 130, wherein antigen-binding domain or the chimeric antigen receptor specifically binds BCMA.

132. The kit of any of claims 129-131, wherein the intracellular signaling domain comprises and intracellular domain of a CD3-zeta (CD3.zeta.) chain.

133. The kit of any of claims 129-132, wherein the chimeric antigen receptor (CAR) further comprises a costimulatory signaling region.

134. The kit of claim 133, wherein the costimulatory signaling region comprises a signaling domain derived from CD28 or 4-1BB, optionally human CD28 or human 4-1BB.

135. The kit of claim 133 or claim 134, wherein the costimulatory signaling region is a domain derived from 4-1BB, optionally human 4-1BB.

136. The kit of any of claims 109-135, wherein the immune cells comprise T cells or NK cells.

137. The kit of claim 136, wherein the immune cells comprise T cells.

138. The kit of claim 136 or claim 137, wherein the T cells are CD4+ and/or CD8+.

139. The kit of any of claims 136-138, wherein the T cells are primary T cells obtained from a subject.

140. An article of manufacture comprising the combination or kit of any of claims 87-139.

141. The method of any of claims 1-86 or the combination or kit of any one of claims 87-139, wherein the disease or condition is a cancer, optionally a multiple myeloma, and administration of the cell therapy and the compound, stereoisomer, pharmaceutically acceptable salt or hydrate results in a decrease in tumor volume, tumor growth, or tumor burden, or an increase in survival or progression-free survival or results in an objective response or complete response in the subject.

142. The method of any of claims 1-86 or the combination or kit of any one of claims 87-139, wherein the disease or condition is a cancer, optionally a multiple myeloma, and the method results in a decrease in tumor volume, tumor growth, or tumor burden, or an increase in survival. progression-free survival, objective response rate, durable response, or complete response rate, in a cohort of subjects, wherein said decrease or increase is greater than an increase or decrease resulting from administration to subjects in the cohort of the cell therapy or of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate alone.

143. The method of any of claims 1-86 or the combination or kit of any one of claims 87-139, wherein the disease or condition is a cancer, optionally a multiple myeloma, and, in a cohort of subjects, the method results in a decrease, as compared to the administration of the cell therapy alone, in tumor volume, tumor growth, or tumor burden, or an increase, as compared to administration of the cell therapy alone, in survival or progression-free survival or objective response rate or complete response rate or measure indicative of cell therapy activity or function.

144. The method of any of 143, wherein the decrease is greater than a decrease in tumor volume, tumor growth, or tumor burden following administration to subjects in the cohort of the compound alone, as compared to no treatment, or wherein the increase is greater than an increase in survival or progression-free survival or objective response rate or complete response rate following administration to subjects in the cohort of the compound alone, as compared to no treatment.

145. The method of any of claims 1-86 or the combination or kit of any one of claims 87-139, wherein the disease or condition is a cancer, optionally a multiple myeloma, and, in a cohort of subjects, the method results in an increase in surface expression of BCMA on the cancer or cancer cells, as compared to the administration of the cell therapy alone.

146. The method of any of claims1-86 or the combination or kit of any one of claims 87-139, wherein the disease or condition is a cancer, optionally a multiple myeloma, and, in a cohort of subjects, the method results in an decrease in serum BCMA levels, as compared to the administration of the cell therapy alone to subjects of the cohort and/or as compared to the administration of the compound alone to subjects of the cohort.

147. The method of claim 146, wherein the decrease as compared to the administration of the cell therapy alone is greater than a decrease in serum BCMA levels in subjects of the cohort following administration of the compound, stereoisomer, pharmaceutically acceptable salt or hydrate alone as compared to no treatment.
Description



CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. provisional patent application 62/582,308, entitled "COMBINATION OF A CELL THERAPY AND A GAMMA SECRETASE INHIBITOR" filed on Nov. 6, 2017; U.S. provisional patent application 62/582,937, entitled "COMBINATION OF A CELL THERAPY AND A GAMMA SECRETASE INHIBITOR" filed on Nov. 7, 2017; and U.S. provisional patent application 62/665,450, entitled "COMBINATION OF A CELL THERAPY AND A GAMMA SECRETASE INHIBITOR" filed on May 1, 2018, the contents of which are incorporated by reference in their entirety.

INCORPORATION BY REFERENCE OF SEQUENCE LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 735042014240SeqList.TXT, created Nov. 6, 2018, which is 320,126 bytes in size. The information in the electronic format of the Sequence Listing is incorporated by reference in its entirety.

FIELD

[0003] The present disclosure relates in some aspects to methods, compositions and uses involving immunotherapies, such as adoptive cell therapy, e.g., T cell therapy, and a gamma secretase inhibitor. The provided methods, compositions and uses include those for combination therapies involving the administration or use of a gamma secretase inhibitor in conjunction with a T cell therapy, such as a genetically engineered T cell therapy involving cells engineered with a recombinant receptor, such as chimeric antigen receptor (CAR)-expressing T cells, such as anti-BCMA chimeric antigen receptor (CAR)-expressing T cells. Also provided are compositions, methods of administration to subjects, articles of manufacture and kits for use in the methods. In some aspects, features of the methods and cells provide for increased or improved activity, efficacy, persistence, expansion and/or proliferation of T cells for adoptive cell therapy or endogenous T cells recruited by immunotherapeutic agents.

BACKGROUND

[0004] Various strategies are available for immunotherapy, for example administering engineered T cells for adoptive therapy. For example, strategies are available for engineering T cells expressing genetically engineered antigen receptors, such as CARs, and administering compositions containing such cells to subjects. Improved strategies are needed to improve efficacy of the cells, for example, improving the persistence, activity and/or proliferation of the cells upon administration to subjects. Provided are methods, compositions, kits, and systems that meet such needs.

SUMMARY

[0005] Provided herein are combination therapies involving administration of an immunotherapy involving a cell therapy, such as a T cell therapy, and an inhibitor of gamma secretase. In some aspects, the provided methods of treating a subject having a disease or disorder involve administration of an immunotherapy or immunotherapeutic agent, such as a composition including cells for adoptive cell therapy, e.g., such as a T cell therapy (e.g. CAR-expressing T cells), and administration of a gamma secretase inhibitor. In some embodiments, the cell therapy, such as CAR-expressing T cells, comprises an antigen-binding domain that binds to a B Cell Maturation Antigen (BCMA), such as surface BCMA. In some embodiments, the method further comprises selecting a subject for treatment, wherein the subject has a low expression of surface BCMA on a cell (such as a tumor/cancer cell). In some embodiments, the cell therapy comprises a recombinant receptor, such as a receptor that does not bind to BCMA. In some embodiments, the gamma secretase inhibitor prohibits intramembrane cleavage of a receptor on a cell (such as a tumor/cancer cell), wherein the cell therapy specifically targets the receptor. In some embodiments, the combination therapy generally involves administration of a gamma secretase inhibitor and administration of a cell therapy, such as a composition including cells for adoptive cell therapy, e.g., such as a T cell therapy (e.g. CAR-expressing T cells), wherein the gamma secretase administration is subsequent to the administration of cell therapy. In some embodiments, the therapy modulates activity of a cell therapy.

[0006] Provided herein are methods of treatment that involve: (a) administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of genetically engineered cells expressing a chimeric antigen receptor (CAR) comprising an antigen-binding domain that specifically binds to surface B cell maturation antigen (BCMA); and (b) administering to the subject an inhibitor of gamma secretase, wherein binding of the CAR to surface BCMA, or a measure indicative of function or activity following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA. In some embodiments, the binding of the CAR to surface BCMA, or a measure indicative of function or activity following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay.

[0007] Provided here are methods of treatment that involve: administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of genetically engineered cells expressing a chimeric antigen receptor comprising an antigen-binding domain that binds to surface B cell maturation antigen (BCMA), wherein binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay, wherein, at the time of initiation of the administration of the cell therapy, the subject has been previously administered, and/or is undergoing treatment with, an inhibitor of gamma secretase.

[0008] Provided here are methods of treatment that involve: administering an inhibitor of gamma secretase to a subject having a disease or disorder, wherein, at the time of initiation of the administration of the inhibitor, the subject has been previously administered, and/or is undergoing treatment with, a cell therapy, said cell therapy comprising a dose of genetically engineered cells expressing a chimeric antigen receptor comprising an antigen-binding domain that specifically binds surface B cell maturation antigen (BCMA), wherein binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay.

[0009] Provided here are methods of treatment that involve: (a) selecting a subject having a cancer in which cells of the cancer in the subject (i) express surface CD138, surface CD38 or a surface plasma cell marker or are derived from plasma cells and (ii) comprise low expression of surface B cell maturation antigen (BCMA) and/or a level of expression of surface BCMA below a threshold level; (b) administering a cell therapy to the subject selected in (a), said cell therapy comprising a dose of genetically engineered cells expressing a chimeric antigen receptor comprising an antigen-binding domain that binds to B cell maturation antigen (BCMA); and (c) administering to the subject an inhibitor of gamma secretase.

[0010] Provided here are methods of treatment that involve: (a) administering an inhibitor of gamma secretase to a subject having a cancer, said subject selected as having plasma cells comprising low expression of surface B cell maturation antigen (BCMA) and/or a level of expression of surface BCMA below a threshold level; and (b) administering to the subject a cell therapy for treating the cancer, said cell therapy comprising a dose of genetically engineered cells expressing a chimeric antigen receptor comprising an antigen-binding domain that specifically binds to B cell maturation antigen (BCMA).

[0011] In some embodiments of any one of the methods provided herein, the threshold level of expression of surface BCMA is lower than the average or median expression or level of surface BCMA on plasma cells in a plurality of control subjects, optionally wherein the control subjects are a group of healthy or normal subjects.

[0012] In some embodiments of any one of the methods provided herein, the low expression of surface BCMA is present when less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma cell marker or phenotype or the cancer cells, in the subject express surface BCMA; or the threshold level of surface BCMA is less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma cell marker or phenotype or the cancer cells, in the subject that express surface BCMA.

[0013] In some embodiments of any one of the methods provided herein, the expression of surface BCMA is determined by flow cytometry and/or an immunoassay.

[0014] In some embodiments of any one of the embodiments provided herein, binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay.

[0015] In some embodiments of any one of the methods provided herein, the antigen-binding domain does not bind soluble BCMA or binds to soluble BCMA with an affinity lower than the affinity of said antigen-binding domain binding to surface BCMA. In some embodiments, the affinity of the antigen-binding domain to the surface B cell maturation antigen (BCMA) is at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold higher than the affinity to the soluble BCMA.

[0016] In some of any of the provided embodiments, the anti-BCMA CAR is an anti-BCMA CAR as described herein. Among a provided anti-BCMA CAR for use in the methods and uses herein is a CAR in which the antibody or antigen-binding fragment contains a V.sub.H region comprising the sequence set forth in SEQ ID NO: 24 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:24; and contains a V.sub.L region comprising the sequence set forth in SEQ ID NO:25 or an amino acid sequence having at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:25. In some embodiments, the antibody or antigen-binding fragment of the provided CAR contains a V.sub.H region that has a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 173, 174 and 175, respectively and a V.sub.L region that has a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively. In some embodiments, the antibody or antigen-binding fragment of the provided CAR contains a V.sub.H region that has a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 176, 177 and 175, respectively and a V.sub.L region that has a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively. In some embodiments, the antibody or antigen-binding fragment of the provided CAR contains a V.sub.H region that has a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 178, 179 and 175, respectively and a V.sub.L region that has a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 183, 184 and 185, respectively. In some embodiments, the antibody or antigen-binding fragment of the provided CAR contains a V.sub.H region that has a CDRH1, a CDRH2 and a CDRH3 comprising the amino acid sequence of SEQ ID NOS: 180, 181 and 182, respectively and a V.sub.L region that has a CDRL1, a CDRL2 and a CDRL3 comprising the amino acid sequence of SEQ ID NOS: 186, 187 and 185, respectively. In some embodiments, the V.sub.H region comprises the sequence set forth in SEQ ID NO:24 and the V.sub.L region comprises the sequence set forth in SEQ ID NO:25. In some embodiments, the antibody or antigen-binding fragment is a single-chain antibody fragment, such as an scFv. In some embodiments, the scFv comprises the sequence of amino acids set forth in SEQ ID NO:188 or a sequence of amino acids at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:188. In some embodiments, the anti-BCMA CAR has the sequence of amino acids set forth in SEQ NO: 124 or a sequence of amino acids at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:124. In some embodiments, the anti-BCMA CAR has the sequence of amino acids set forth in SEQ NO: 125 or a sequence of amino acids at least at or about 90%, at or about 91%, at or about 92%, at or about 93%, at or about 94%, at or about 95%, at or about 96%, at or about 97%, at or about 98%, or at or about 99% identity to SEQ ID NO:125.

[0017] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor inhibits intramembrane cleavage of BCMA.

[0018] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor inhibits intramembrane cleavage of BCMA with a half-maximal inhibitory concentration (IC.sub.50) of 0.01 nM to 10 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.01 nM to 0.5 nM, 0.01 nM to 0.35 nM, 0.01 nM to 0.25 nM, 0.01 nM to 0.1 nM, 0.01 nM to 0.05 nM, 0.05 nM to 10 nM, 0.05 nM to 5 nM, 0.05 nM to 1 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.35 nM, 0.05 nM to 0.25 nM, 0.05 nM to 0.1 nM, 0.1 nM to 10 nM, 0.1 nM to 5 nM, 0.1 nM to 1 nM, 0.1 nM, to 0.5 nM, 0.1 nM to 0.35 nM, 0.1 nM to 0.25 nM, 0.25 nM to 10 nM, 0.25 nM to 5 nM, 0.25 nM to 1 nM, 0.25 nM to 0.5 nM, 0.25 nM to 0.35 nM, 0.35 nM to 10 nM, 0.35 nM to 5 nM, 0.35 nM to 1 nM, 0.35 nM to 0.5 nM, 0.5 nM to 10 nM, 0.5 nM to 5 nM, 0.5 nM to 1 nM, 1 nM to 10 nM, 1 nM to 5 nM or 5 nM to 10 nM.

[0019] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor inhibits intramembrane cleavage of BCMA with a half-maximal inhibitory concentration (IC.sub.50) of less than or less than about 10 nM, 5 nM, 1 nM, 0.5 nM, 0.35 nM, 0.25 nM, 0.1 nM, 0.05 nM or 0.01 nM or less.

[0020] Provided herein are methods of treatment that involve: (a) administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor, wherein the recombinant receptor does not specifically bind to surface B cell maturation antigen (BCMA); and (b) administering an inhibitor of gamma secretase.

[0021] Provided herein are methods of treatment that involve: administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor, wherein the recombinant receptor does not specifically bind to surface B cell maturation antigen (BCMA), wherein, at the time of initiation of the administration of the cell therapy, the subject has been previously administered, and/or is undergoing treatment with, an inhibitor of gamma secretase.

[0022] Provided herein are methods of treatment that involve: administering an inhibitor of gamma secretase to a subject having a disease or disorder, wherein, at the time of initiation of the administration of the inhibitor, the subject has been previously administered, and/or is undergoing treatment with, a cell therapy, said cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor, wherein the recombinant receptor does not specifically bind to surface B cell maturation antigen (BCMA).

[0023] In some embodiments of any one of the methods provided herein, the recombinant receptor specifically binds to a target antigen associated with the disease or disorder.

[0024] In some embodiments of any one of the methods provided herein, the target antigen is selected from among, carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, a pathogen-specific antigen and an antigen associated with a universal tag.

[0025] Provided herein are methods of treatment that involve: (a) administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor; and (b) subsequent to the administration in (a) administering to the subject an inhibitor of gamma secretase.

[0026] Provided herein are methods of treatment that involve: administering an inhibitor of gamma secretase to a subject having a disease or disorder, wherein, at the time of initiation of the administration of the inhibitor, the subject has been previously administered, and/or is undergoing treatment with, a cell therapy, said cell therapy comprising a dose of genetically engineered cells expressing recombinant receptor.

[0027] In some embodiments of any one of the methods provided herein, the recombinant receptor specifically binds to a target antigen associated with the disease or disorder.

[0028] In some embodiments of any one of the methods provided herein, the target antigen is selected from among, B cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, a pathogen-specific antigen and an antigen associated with a universal tag. In some embodiments, the target antigen is Muc1. In some embodiments of any one of the methods provided herein, the target antigen is BCMA. In some embodiments, wherein the BCMA is surface BCMA.

[0029] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor inhibits or reduces or is capable of inhibiting or reducing cleavage of one or more targets selected from BCMA, Notch 1, Notch 2, Notch 3, Notch 4, Muc1, Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CXCR1, CXCL16, Delta1, E-cadherein, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R or ameloid precursor protein (APP). In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises Muc1. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises BCMA.

[0030] In some embodiments of any one of the methods provided herein, cleavage of one or more targets selected from BCMA, Notch 1, Notch 2, Notch 3, Notch 4, Muc1, Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CXCR1, CXCL16, Delta1, E-cadherein, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R or ameloid precursor protein (APP) is inhibited or reduced or can be inhibited or reduced by the gamma secretase inhibitor. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises Muc1. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises BCMA.

[0031] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor inhibits the target with a half-maximal inhibitory concentration (IC.sub.50) of 0.01 nM to 1 .mu.M, 0.01 nM to 100 nM, 0.01 nM to 10 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.01 nM to 0.5 nM, 0.01 nM to 0.25 nM, 0.01 nM to 0.1 nM, 0.01 nM to 0.05 nM, 0.05 nM to 1 .mu.M, 0.05 nM to 100 nM, 0.05 nM to 10 nM, 0.05 nM to 5 nM, 0.05 nM to 1 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.25 nM, 0.05 nM to 0.1 nM, 0.1 nM to 1 .mu.M, 0.1 nM to 100 nM, 0.1 nM to 10 nM, 0.1 nM to 5 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.1 nM to 0.25 nM, 0.25 nM to 1 .mu.M, 0.25 nM to 100 nM, 0.25 nM to 10 nM, 0.25 nM to 5 nM, 0.25 nM to 1 nM, 0.25 nM to 0.5 nM, 0.5 nM to 1 .mu.M, 0.5 nM to 100 nM, 0.5 nM to 10 nM, 0.5 nM to 5 nM, 0.5 nM to 1 nM, 1 nM to 1 .mu.M, 1 nM to 100 nM, 1 nM to 10 nM, 1 nM to 5 nM, 5 nM to 1 .mu.M, 5 nM to 100 nM, 5 nM to 10 nM, 10 nM to 1 .mu.M, 10 nM to 100 nM or 100 nM to 1 .mu.M.

[0032] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor inhibits the target with a half-maximal inhibitory concentration (IC.sub.50) of less than or less than about 1 .mu.M, 100 nM, 10 nM, 5 nM, 1 nM, 0.5 nM, 0.35 nM, 0.25 nM, 0.1 nM, 0.05 nM or 0.01 nM or less.

[0033] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor is a peptide inhibitor or non-peptide inhibitor. In some embodiments, the gamma secretase inhibitor is a peptide inhibitor and the peptide inhibitor is selected from among peptide aldehydes derivatives, difluoroketones derivatives, hydroxyethylene dipeptide isotere derivatives, alpha-helical peptide derivatives and dipeptide analogs. In some embodiments, the gamma secretase inhibitor is a non-peptide inhibitor and the non-peptide inhibitor is a benzodiazepines derivative or a sulfonamides derivative.

[0034] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor is a transition state inhibitor or non-transition state inhibitor.

[0035] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor is a nonsteroidal anti-inflammatory drug.

[0036] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor is selected from LY3039478, secretase inhibitor I (GSI I) Z-Leu-Leu-Norleucine; .gamma.-secretase inhibitor II (GSI II); .gamma.-secretase inhibitor III (GSI III), N-Benzyloxycarbonyl-Leu-leucinal, N-(2-Naphthoyl)-Val-phenylalaninal; .gamma.-secretase inhibitor III (GSI IV); .gamma.-secretase inhibitor III (GSI V), N-Benzyloxycarbonyl-Leu-phenylalaninal; .gamma.-secretase inhibitor III (GSI VI), 1-(S)-endo-N-(1,3,3)-Trimethylbicyclo[2.2.1]hept-2-yl)-4-fluorophenyl Sulfonamide; .gamma.-secretase inhibitor III (GSI VII), Menthyloxycarbonyl-LL-CHO; .gamma.-secretase inhibitor III (GSI IX), (DAPT), N-[N-(3,5-Difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester; .gamma.-secretase inhibitor X (GSI X), {1S-Benzyl-4R-[1-(1S-carbamoyl-2-phenethylcarbamoyl)-1S-3-methylbutylcarb- -amoyl]-2R-hydroxy-5-phenylpentyl}carbamic Acid tert-butyl Ester; .gamma.-secretase inhibitor XI (GSI XI), 7-Amino-4-chloro-3-methoxyisocoumarin; .gamma.-secretase inhibitor XII (GSI XII), Z-Ile-Leu-CHO; .gamma.-secretase inhibitor XIII (GSI XIII), Z-Tyr-Ile-Leu-CHO; .gamma.-secretase inhibitor XIV (GSI XIV), Z-Cys(t-Bu)-Ile-Leu-CHO; .gamma.-secretase inhibitor XVI (GSI XVI), N-[N-3,5-Difluorophenacetyl]-L-alanyl-S-phenylglycine Methyl Ester; .gamma.-secretase inhibitor XVII (GSI XVII); .gamma.-secretase inhibitor XIX (GSI XIX), benzo[e][1,4]diazepin-3-yl)-butyramide; .gamma.-secretase inhibitor XX (GSI XX), (S,S)-2-[2-(3,5-Difluorophenyl)acetylamino]-N-(5-methyl-6-oxo-6,7-dihydro- -5H-dibenzo[b,d]azepin-7-yl)propionamide; .gamma.-secretase inhibitor XXI (GSI XXI), (S,S)-2-[2-(3,5-Difluorophenyl)-acetylamino]-N-(1-methyl-2-oxo-5-phenyl-2- -,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-propionamide; Gamma40 secretase inhibitor I, N-trans-3,5-Dimethoxycinnamoyl-Ile-leucinal; Gamma40 secretase inhibitor II, N-tert-Butyloxycarbonyl-Gly-Val-Valinal Isovaleryl-V V-Sta-A-Sta-OCH3; MK-0752; MRK-003 (Merck); semagacestat/LY450139; RO4929097; PF-03084,014; BMS-708163; MPC-7869 (.gamma.-secretase modifier), YO-01027 (Dibenzazepine), Compound E ([(2S)-2-{[(3,5-Difluorophenyl)acetyl]amino}-N-[(3S)-1-methyl-2-oxo-5-phe- nyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide], LY411575, L-685,458, BMS-289948 (4-chloro-N-(2,5-difluorophenyl)-N-((1R)-{4-fluoro-2-[3-(1H-imidazol-1-yl- )propyl]phenyl}ethyl)benzenesulfonamide hydrochloride) and BMS-299897 (4-[2-((1R)-1-{[(4-chlorophenyl)sulfonyl]-2,5-difluoroanilino}ethyl)-5-fl- uorophenyljbutanoic acid).

[0037] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor is a compound of the structure:

##STR00001##

[0038] or a pharmaceutically acceptable salt of hydrate thereof.

[0039] Provided herein are methods of treatment that involve: (a) administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor; and (b) administering to the subject a compound of the structure:

##STR00002##

[0040] or a pharmaceutically acceptable salt of hydrate thereof.

[0041] Provided herein are methods of treatment that involve: administering a cell therapy to a subject having a disease or disorder, said cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor, wherein, at the time of initiation of the administration of the cell therapy, the subject has been previously administered, and/or is undergoing treatment with, a compound of the structure:

##STR00003##

[0042] or a pharmaceutically acceptable salt of hydrate thereof.

[0043] Provided herein are methods of treatment that involve: administering to a subject having a disease or disorder a compound of the structure:

##STR00004##

[0044] or a pharmaceutically acceptable salt of hydrate thereof, wherein, at the time of initiation of the administration, the subject has been previously administered, and/or is undergoing treatment with a cell therapy, said cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor.

[0045] In some embodiments of any one of the methods provided herein, the recombinant receptor specifically binds to a target antigen associated with the disease or disorder. In some embodiments, the target antigen is selected from among, B cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, a pathogen-specific antigen and an antigen associated with a universal tag. In some embodiments, the target antigen is Muc1. In some embodiments, the target antigen is BCMA. In some embodiments, the BCMA is surface BCMA. In some embodiments, the recombinant receptor does not bind soluble BCMA or binds to soluble BCMA with an affinity lower than the affinity of said recombinant receptor for binding to surface BCMA. In some embodiments, the affinity of the antigen-binding domain to the surface B cell maturation antigen (BCMA) is at least 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold higher than the affinity to the soluble BCMA.

[0046] In some embodiments of any one of the methods provided herein, the subject comprises plasma cells, or cancer cells or myeloma cells or cells expressing plasma cell markers, expressing surface BCMA.

[0047] In some embodiments of any one of the methods provided herein, the subject is selected as having cells comprising low expression of surface B cell maturation antigen (BCMA); and/or selecting a subject for administration of the cell therapy and/or the inhibitor based on having plasma cells comprising low expression of surface BCMA.

[0048] In some embodiments of any one of the methods provided herein, the target of the gamma secretase inhibitor is the target antigen and the gamma secretase inhibitor inhibits cleavage of the target antigen.

[0049] In some embodiments of any one of the methods provided herein, administration of the inhibitor: decreases BCMA cleavage or shedding from cells, optionally plasma cells, by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of BCMA cleavage or shedding from cells in the subject prior to administration of the inhibitor; decreases the level or amount of BCMA detected in the serum of a subject by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level or amount of BCMA in the serum of the subject prior to administration of the inhibitor; and/or increases expression of surface BCMA on cells, optionally plasma cells by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of surface BCMA on the cells in the subject prior to administration of the inhibitor.

[0050] In some embodiments of any one of the methods provided herein, administration of the inhibitor: decreases cleavage or shedding of the target or the target antigen, optionally BCMA or Muc1, from cells, optionally plasma cells, by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of cleavage or shedding of the target or target antigen from cells in the subject prior to administration of the inhibitor; decreases the level or amount of the target or target antigen, optionally BCMA or Muc1, detected in the serum of a subject by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level or amount of the target or target antigen in the serum of the subject prior to administration of the inhibitor; and/or increases expression of surface target or target antigen, optionally BCMA or Muc1, on cells, optionally plasma cells, by greater than or greater than about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more compared to the level of surface target or target antigen on the cells in the subject prior to administration of the inhibitor.

[0051] In some embodiments of any one of the methods provided herein, the disease or disorder is a cancer. In some embodiments, the cancer is a B cell malignancy. In some embodiments, the cancer is multiple myeloma, plasmacytoma, a cancer of plasma cell origin and/or a cancer of B cell origin.

[0052] In some embodiments of any one of the methods provided herein, the inhibitor is administered orally, subcutaneously or intravenously. In some embodiments, the inhibitor is administered orally.

[0053] In some embodiments of any one of the methods provided herein, the inhibitor is administered at least or is administered six times daily, five times daily, four times daily, three times daily, twice daily, once daily, every other day, three times a week, at least once a week, or only one time. In some embodiments, the inhibitor is administered three times a week.

[0054] In some embodiments of any one of the methods provided herein, the administration of the inhibitor is carried out in a treatment cycle that is at least or at least about or 14 days, at least or at least about or 21 days or at least or at least about or 28 days.

[0055] In some embodiments of any one of the methods provided herein, the inhibitor is administered, or each administration of the inhibitor is independently administered, in an amount of 0.5 mg to 500 mg, 0.5 mg to 250 mg, 0.5 mg to 100 mg, 0.5 mg to 50 mg, 0.5 mg to 25 mg, 0.5 mg to 10 mg, 0.5 mg to 5.0 mg, 0.5 mg to 2.5 mg, 0.5 mg to 1.0 mg, 1.0 mg to 500 mg, 1.0 mg to 250 mg, 1.0 mg to 100 mg, 1.0 mg to 50 mg, 1.0 mg to 25 mg, 1.0 mg to 10 mg, 1.0 mg to 5.0 mg, 1.0 mg to 2.5 mg, 2.5 mg to 500 mg, 2.5 mg to 250 mg, 2.5 mg to 100 mg, 2.5 mg to 50 mg, 2.5 mg to 25 mg, 2.5 mg to 10 mg, 2.5 mg to 5.0 mg, 5.0 mg to 500 mg, 5.0 mg to 250 mg, 5.0 mg to 100 mg, 5.0 mg to 50 mg, 5.0 mg to 25 mg, 5.0 mg to 10 mg, 10 mg to 500 mg, 10 mg to 250 mg, 10 mg to 100 mg, 10 mg to 50 mg, 10 mg to 25 mg, 25 mg to 500 mg, 25 mg to 250 mg, 25 mg to 100 mg, 25 mg to 50 mg, 50 mg to 500 mg, 50 mg to 250 mg, 50 mg to 100 mg, 100 mg to 500 mg, 100 mg to 250 mg or 250 mg to 500 mg. In some embodiments, the inhibitor is administered, or each administration of the inhibitor is independently administered, in an amount that is at least or at least about or is or is about 0.5 mg, 1.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 25 mg, 50 mg, 100 mg, 250 mg or 500 mg.

[0056] In some embodiments of any one of the methods provided herein, the recombinant receptor is a transgenic T cell receptor (TCR) or a functional non-T cell receptor. In some embodiments, the recombinant receptor is a chimeric receptor, which optionally is a chimeric antigen receptor (CAR). In some embodiments, the chimeric antigen receptor (CAR) comprises an extracellular antigen-recognition domain that specifically binds to the antigen and an intracellular signaling domain comprising an ITAM. In some embodiments, the intracellular signaling domain comprises and intracellular domain of a CD3-zeta (CD3.zeta.) chain. In some embodiments, the chimeric antigen receptor (CAR) further comprises a costimulatory signaling region. In some embodiments, the costimulatory signaling region comprises a signaling domain derived from CD28 or 4-1BB, optionally human CD28 or human 4-1BB. In some embodiments, the costimulatory signaling region is a domain derived from 4-1BB, optionally human 4-1BB.

[0057] In some embodiments of any one of the methods provided herein, the subject is a human.

[0058] In some embodiments of any one of the methods provided herein, the BCMA is human BCMA or the target antigen is a human antigen.

[0059] In some embodiments of any one of the methods provided herein, the genetically engineered cells comprise T cells or NK cells. In some embodiments, the cell therapy is a T cell therapy and the dose of genetically engineered cells comprises T cells. In some embodiments, the T cells are CD4+ and/or CD8+. In some embodiments, the T cells are primary T cells obtained from a subject.

[0060] In some embodiments of any one of the methods provided herein, the cell therapy comprises cells that are autologous to the subject.

[0061] In some embodiments of any one of the methods provided herein, the cell therapy comprises cells that are allogeneic to the subject.

[0062] In some embodiments of any one of the methods provided herein, the cell therapy comprises the administration of from or from about 1.times.10.sup.5 to 5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs); the cell therapy comprises the administration of from or from about 1.times.10.sup.5 to 1.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total PBMCs; from or from about 5.times.10.sup.5 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs; or from or from about 1.times.10.sup.6 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs, each inclusive.

[0063] In some embodiments of any one of the methods provided herein, the cell therapy comprises the administration of no more than 5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs); no more than 2.5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 1.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 0.5.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 1.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 0.5.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total PBMCs.

[0064] In some embodiments of any one of the methods provided herein, the initiation of administration of the inhibitor is prior to, concurrently with or subsequently to initiation of administration of the cell therapy. In some embodiments, the inhibitor is administered prior to initiation of administration of the cell therapy. In some embodiments, initiation of administration of the inhibitor is within, or within about, 1 hours, 2 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours or 1 week prior to the initiation of the administration of the cell therapy. In some embodiments, the inhibitor is administered subsequently to initiation of administration of the cell therapy. In some embodiments, initiation of administration of the inhibitor is within, or within about, 1 hours, 2 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours,1 week, 14 days, 21 days, 28 days or more after the initiation of the administration of the cell therapy. In some embodiments, the inhibitor is administered up to 7 days, 14 days, 21 days, 28 days or more after initiation of the administration of the cells.

[0065] In some embodiments of any one of the methods provided herein, the inhibitor is administered at a time in which: the number of cells of the cell therapy detectable in the blood from the subject is decreased compared to in the subject at a preceding time point after initiation of the administration of the cells; the number of cells of the cell therapy detectable in the blood is less than or less than about 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 50-fold or 100-fold or less the peak or maximum number of the cells of the cell therapy detectable in the blood of the subject after initiation of administration of the administration of the cells; and/or at a time after a peak or maximum level of the cells of the cell therapy are detectable in the blood of the subject, the number of cells of or derived from the cells detectable in the blood from the subject is less than less than 10%, less than 5%, less than 1% or less than 0.1% of total peripheral blood mononuclear cells (PBMCs) in the blood of the subject.

[0066] In some embodiments of any one of the methods provided herein, the method further comprises administering a lymphodepleting chemotherapy prior to administration of the cell therapy and/or wherein the subject has received a lymphodepleting chemotherapy prior to administration of the cells. In some embodiments, the lymphodepleting chemotherapy comprises administering fludarabine and/or cyclophosphamide to the subject.

[0067] In some embodiments of any one of the methods provided herein, the method further comprises administering a steroid, optionally wherein the steroid is administered prior to, concurrently with and/or subsequently to initiation of administration of the inhibitor, optionally wherein the steroid is administered during the cycle of treatment with the inhibitor. In some embodiments, the steroid is or comprises dexamethasone.

[0068] In some embodiments of any one of the methods provided herein, the cell therapy exhibits increased or prolonged expansion and/or persistence in the subject as compared to a method in which the cell therapy is administered to the subject in the absence of the gamma secretase inhibitor.

[0069] In some embodiments of any one of the methods provided herein, the method thereby prevents, reduces or ameliorates one or more symptoms or outcomes of the disease or disorder.

[0070] Provided herein are methods of treatment that involve: (a) administering to a subject having a disease or disorder, a therapeutic agent or therapy that targets or is specific for B cell maturation antigen (BCMA); and (b) administering a gamma secretase inhibitor to the subject.

[0071] In some embodiments of any one of the methods provided herein, the therapeutic agent or therapy is or comprises an antibody or antigen-binding fragment thereof, optionally a bispecific antibody. In some embodiments, therapeutic agent or therapy is a bispecific antibody that further targets or specifically binds to a T cell antigen, optionally CD2 or CD3. In some embodiments, the therapeutic agent or therapy is a bispecific antibody that further targets a second antigen, optionally wherein the second antigen is selected from CD19, CD20, CD22, CD33, CD38, CS1, ROR1, GPC3, CD123, IL-13R, CD138, c-Met, EGFRvIII, GD-2, NY-ESO-1, MAGE A3, and glycolipid F77.

[0072] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor is selected from LY3039478, secretase inhibitor I (GSI I) Z-Leu-Leu-Norleucine; .gamma.-secretase inhibitor II (GSI II); .gamma.-secretase inhibitor III (GSI III), N-Benzyloxycarbonyl-Leu-leucinal, N-(2-Naphthoyl)-Val-phenylalaninal; .gamma.-secretase inhibitor III (GSI IV); .gamma.-secretase inhibitor III (GSI V), N-Benzyloxycarbonyl-Leu-phenylalaninal; .gamma.-secretase inhibitor III (GSI VI), 1-(S)-endo-N-(1,3,3)-Trimethylbicyclo[2.2.1]hept-2-yl)-4-fluorophenyl Sulfonamide; .gamma.-secretase inhibitor III (GSI VII), Menthyloxycarbonyl-LL-CHO; .gamma.-secretase inhibitor III (GSI IX), (DAPT), N-[N-(3,5-Difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester; .gamma.-secretase inhibitor X (GSI X), {1S-Benzyl-4R-[1-(1S-carbamoyl-2-phenethylcarbamoyl)-1S-3-methylbutylcarb- -amoyl]-2R-hydroxy-5-phenylpentyl}carbamic Acid tert-butyl Ester; .gamma.-secretase inhibitor XI (GSI XI), 7-Amino-4-chloro-3-methoxyisocoumarin; .gamma.-secretase inhibitor XII (GSI XII), Z-Ile-Leu-CHO; .gamma.-secretase inhibitor XIII (GSI XIII), Z-Tyr-Ile-Leu-CHO; .gamma.-secretase inhibitor XIV (GSI XIV), Z-Cys(t-Bu)-Ile-Leu-CHO; .gamma.-secretase inhibitor XVI (GSI XVI), N-[N-3,5-Difluorophenacetyl]-L-alanyl-S-phenylglycine Methyl Ester; .gamma.-secretase inhibitor XVII (GSI XVII); .gamma.-secretase inhibitor XIX (GSI XIX), benzo[e][1,4]diazepin-3-yl)-butyramide; .gamma.-secretase inhibitor XX (GSI XX), (S,S)-2-[2-(3,5-Difluorophenyl)acetylamino]-N-(5-methyl-6-oxo-6,7-dihydro- -5H-dibenzo[b,d]azepin-7-yl)propionamide; .gamma.-secretase inhibitor XXI (GSI XXI), (S,S)-2-[2-(3,5-Difluorophenyl)-acetylamino]-N-(1-methyl-2-oxo-5-phenyl-2- -,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-propionamide; Gamma40 secretase inhibitor I, N-trans-3,5-Dimethoxycinnamoyl-Ile-leucinal; Gamma40 secretase inhibitor II, N-tert-Butyloxycarbonyl-Gly-Val-Valinal Isovaleryl-V V-Sta-A-Sta-OCH3; MK-0752; MRK-003 (Merck); semagacestat/LY450139; RO4929097; PF-03084,014; BMS-708163; MPC-7869 (.gamma.-secretase modifier), YO-01027 (Dibenzazepine), Compound E ([(2S)-2-{[(3,5-Difluorophenyl)acetyl]amino}-N-[(3S)-1-methyl-2-oxo-5-phe- nyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide], LY411575, L-685,458, BMS-289948 (4-chloro-N-(2,5-difluorophenyl)-N-((1R)-{4-fluoro-2-[3-(1H-imidazol-1-yl- )propyl]phenyl}ethyl)benzenesulfonamide hydrochloride) and BMS-299897 (4-[2-((1R)-1-{[(4-chlorophenyl)sulfonyl]-2,5-difluoroanilino}ethyl)-5-fl- uorophenyljbutanoic acid).

[0073] In some embodiments of any one of the methods provided herein, the gamma secretase inhibitor is a compound of the structure:

##STR00005##

[0074] or a pharmaceutically acceptable salt of hydrate thereof.

[0075] Provided herein are combinations that include: (a) genetically engineered cells expressing a chimeric receptor comprising an antigen binding domain that binds to surface B cell maturation antigen (BCMA); (b) an inhibitor of gamma secretase, wherein binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay.

[0076] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor inhibits intramembrane cleavage of BCMA.

[0077] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor inhibits intramembrane cleavage of BCMA with a half-maximal inhibitory concentration (IC.sub.50) of 0.01 nM to 10 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.01 nM to 0.5 nM, 0.01 nM to 0.35 nM, 0.01 nM to 0.25 nM, 0.01 nM to 0.1 nM, 0.01 nM to 0.05 nM, 0.05 nM to 10 nM, 0.05 nM to 5 nM, 0.05 nM to 1 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.35 nM, 0.05 nM to 0.25 nM, 0.05 nM to 0.1 nM, 0.1 nM to 10 nM, 0.1 nM to 5 nM, 0.1 nM to 1 nM, 0.1 nM, to 0.5 nM, 0.1 nM to 0.35 nM, 0.1 nM to 0.25 nM, 0.25 nM to 10 nM, 0.25 nM to 5 nM, 0.25 nM to 1 nM, 0.25 nM to 0.5 nM, 0.25 nM to 0.35 nM, 0.35 nM to 10 nM, 0.35 nM to 5 nM, 0.35 nM to 1 nM, 0.35 nM to 0.5 nM, 0.5 nM to 10 nM, 0.5 nM to 5 nM, 0.5 nM to 1 nM, 1 nM to 10 nM, 1 nM to 5 nM or 5 nM to 10 nM. In some embodiments, the gamma secretase inhibitor inhibits intramembrane cleavage of BCMA with a half-maximal inhibitory concentration (IC.sub.5o) of less than or less than about 10 nM, 5 nM, 1 nM., 0.5 nM, 0.35 nM, 0.25 nM, 0.1 nM, 0.05 nM or 0.01 nM or less.

[0078] Provided herein are combinations that include: (a) genetically engineered cells expressing a recombinant receptor, wherein the recombinant receptor does not specifically bind to surface B cell maturation antigen (BCMA); and (b) an inhibitor of gamma secretase.

[0079] Provided herein are combinations that include: (a) genetically engineered cells expressing a recombinant receptor, wherein the recombinant receptor does not specifically bind to surface B cell maturation antigen (BCMA); and (b) an inhibitor of gamma secretase, wherein binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay.

[0080] In some embodiments of any one of the combinations provided herein, the recombinant receptor specifically binds to a target antigen associated with the disease or disorder. In some embodiments, the target antigen is selected from among, carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, a pathogen-specific antigen and an antigen associated with a universal tag. In some embodiments, the target antigen is Muc1.

[0081] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor inhibits or reduces or is capable of inhibiting or reducing cleavage of one or more targets selected from BCMA, Notch 1, Notch 2, Notch 3, Notch 4, Muc1, Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CXCR1, CXCL16, Delta1, E-cadherein, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R or ameloid precursor protein (APP). In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises Muc1. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises BCMA.

[0082] In some embodiments of any one of the combinations provided herein, cleavage of one or more targets selected from BCMA, Notch 1, Notch 2, Notch 3, Notch 4, Muc1, Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CXCR1, CXCL16, Delta1, E-cadherein, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R or ameloid precursor protein (APP) is inhibited or reduced or can be inhibited or reduced by the gamma secretase inhibitor. In some embodiments, the one or more targets the cleavage of which is inhibited or reduced comprises Muc1. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises BCMA.

[0083] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor inhibits the target with a half-maximal inhibitory concentration (IC.sub.50) of 0.01 nM to 1 .mu.M, 0.01 nM to 100 nM, 0.01 nM to 10 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.01 nM to 0.5 nM, 0.01 nM to 0.25 nM, 0.01 nM to 0.1 nM, 0.01 nM to 0.05 nM, 0.05 nM to 1 .mu.M, 0.05 nM to 100 nM, 0.05 nM to 10 nM, 0.05 nM to 5 nM, 0.05 nM to 1 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.25 nM, 0.05 nM to 0.1 nM, 0.1 nM to 1 .mu.M, 0.1 nM to 100 nM, 0.1 nM to 10 nM, 0.1 nM to 5 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.1 nM to 0.25 nM, 0.25 nM to 1 .mu.M, 0.25 nM to 100 nM, 0.25 nM to 10 nM, 0.25 nM to 5 nM, 0.25 nM to 1 nM, 0.25 nM to 0.5 nM, 0.5 nM to 1 .mu.M, 0.5 nM to 100 nM, 0.5 nM to 10 nM, 0.5 nM to 5 nM, 0.5 nM to 1 nM, 1 nM to 1 .mu.M, 1 nM to 100 nM, 1 nM to 10 nM, 1 nM to 5 nM, 5 nM to 1 .mu.M, 5 nM to 100 nM, 5 nM to 10 nM, 10 nM to 1 .mu.M, 10 nM to 100 nM or 100 nM to 1 .mu.M. In some embodiments, the gamma secretase inhibitor inhibits the target with a half-maximal inhibitory concentration (IC.sub.50) of less than or less than about 1 .mu.M, 100 nM, 10 nM, 5 nM, 1 nM, 0.5 nM, 0.35 nM, 0.25 nM, 0.1 nM, 0.05 nM or 0.01 nM or less.

[0084] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor is a peptide inhibitor or non-peptide inhibitor. In some embodiments, the gamma secretase inhibitor is a peptide inhibitor and the peptide inhibitor is selected from among peptide aldehydes derivatives, difluoroketones derivatives, hydroxyethylene dipeptide isotere derivatives, alpha-helical peptide derivatives and dipeptide analogs. In some embodiments, the gamma secretase inhibitor is a non-peptide inhibitor and the non-peptide inhibitor is a benzodiazepines derivative or a sulfonamides derivative.

[0085] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor is a transition state inhibitor or non-transition state inhibitor.

[0086] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor is a nonsteroidal anti-inflammatory drug.

[0087] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor is selected from LY3039478, secretase inhibitor I (GSI I) Z-Leu-Leu-Norleucine; .gamma.-secretase inhibitor II (GSI II); .gamma.-secretase inhibitor III (GSI III), N-Benzyloxycarbonyl-Leu-leucinal, N-(2-Naphthoyl)-Val-phenylalaninal; .gamma.-secretase inhibitor III (GSI IV); .gamma.-secretase inhibitor III (GSI V), N-Benzyloxycarbonyl-Leu-phenylalaninal; .gamma.-secretase inhibitor III (GSI VI), 1-(S)-endo-N-(1,3,3)-Trimethylbicyclo[2.2.1]hept-2-yl)-4-fluorophenyl Sulfonamide; .gamma.-secretase inhibitor III (GSI VII), Menthyloxycarbonyl-LL-CHO; .gamma.-secretase inhibitor III (GSI IX), (DAPT), N-[N-(3,5-Difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester; .gamma.-secretase inhibitor X (GSI X), {1S-Benzyl-4R-[1-(1S-carbamoyl-2-phenethylcarbamoyl)-1S-3-methylbutylcarb- -amoyl]-2R-hydroxy-5-phenylpentyl}carbamic Acid tert-butyl Ester; .gamma.-secretase inhibitor XI (GSI XI), 7-Amino-4-chloro-3-methoxyisocoumarin; .gamma.-secretase inhibitor XII (GSI XII), Z-Ile-Leu-CHO; .gamma.-secretase inhibitor XIII (GSI XIII), Z-Tyr-Ile-Leu-CHO; .gamma.-secretase inhibitor XIV (GSI XIV), Z-Cys(t-Bu)-Ile-Leu-CHO; .gamma.-secretase inhibitor XVI (GSI XVI), N-[N-3,5-Difluorophenacetyl]-L-alanyl-S-phenylglycine Methyl Ester; .gamma.-secretase inhibitor XVII (GSI XVII); .gamma.-secretase inhibitor XIX (GSI XIX), benzo[e][1,4]diazepin-3-yl)-butyramide; .gamma.-secretase inhibitor XX (GSI XX), (S,S)-2-[2-(3,5-Difluorophenyl)acetylamino]-N-(5-methyl-6-oxo-6,7-dihydro- -5H-dibenzo[b,d]azepin-7-yl)propionamide; .gamma.-secretase inhibitor XXI (GSI XXI), (S,S)-2-[2-(3,5-Difluorophenyl)-acetylamino]-N-(1-methyl-2-oxo-5-phenyl-2- -,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-propionamide; Gamma40 secretase inhibitor I, N-trans-3,5-Dimethoxycinnamoyl-Ile-leucinal; Gamma40 secretase inhibitor II, N-tert-Butyloxycarbonyl-Gly-Val-Valinal Isovaleryl-V V-Sta-A-Sta-OCH3; MK-0752; MRK-003 (Merck); semagacestat/LY450139; RO4929097; PF-03084,014; BMS-708163; MPC-7869 (.gamma.-secretase modifier), YO-01027 (Dibenzazepine), Compound E ([(2S)-2-{[(3,5-Difluorophenyl)acetyl]amino}-N-[(3S)-1-methyl-2-oxo-5-phe- nyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide], LY411575, L-685,458, BMS-289948 (4-chloro-N-(2,5-difluorophenyl)-N-((1R)-{4-fluoro-2-[3-(1H-imidazol-1-yl- )propyl]phenyl}ethyl)benzenesulfonamide hydrochloride) and BMS-299897 (4-[2-((1R)-1-{[(4-chlorophenyl)sulfonyl]-2,5-difluoroanilino}ethyl)-5-fl- uorophenyljbutanoic acid).

[0088] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor is a compound of the structure:

##STR00006##

[0089] or a pharmaceutically acceptable salt of hydrate thereof.

[0090] Provided herein are combinations that include: (a) genetically engineered cells expressing a recombinant receptor; and (b) a compound of the structure:

##STR00007##

[0091] or a pharmaceutically acceptable salt of hydrate thereof.

[0092] In some embodiments of any one of the combinations provided herein, the recombinant receptor specifically binds to a target antigen associated with the disease or disorder. In some embodiments, the target antigen is selected from among, B cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, a pathogen-specific antigen and an antigen associated with a universal tag. In some embodiments, the target antigen is Muc1.

[0093] In some embodiments of any one of the combinations provided herein, the target antigen is BCMA. In some embodiments, the BCMA is surface BCMA. In some embodiments, the binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay. In some embodiments, the recombinant receptor does not bind soluble BCMA or binds to soluble BCMA with an affinity lower than the affinity of said recombinant receptor for binding to surface BCMA.

[0094] In some embodiments of any one of the combinations provided herein, the recombinant receptor is a transgenic T cell receptor (TCR) or a functional non-T cell receptor. In some embodiments, the recombinant receptor is a chimeric receptor, which optionally is a chimeric antigen receptor (CAR). In some embodiments, the chimeric antigen receptor (CAR) comprises an extracellular antigen-recognition domain that specifically binds to the antigen and an intracellular signaling domain comprising an ITAM. In some embodiments, the intracellular signaling domain comprises and intracellular domain of a CD3-zeta (CD3.zeta.) chain. In some embodiments, the chimeric antigen receptor (CAR) further comprises a costimulatory signaling region. In some embodiments, the costimulatory signaling region comprises a signaling domain derived from CD28 or 4-1BB, optionally human CD28 or human 4-1BB. In some embodiments, the costimulatory signaling region is a domain derived from 4-1BB, optionally human 4-1BB.

[0095] In some embodiments of any one of the combinations provided herein, the BCMA is human BCMA or the target antigen is a human antigen.

[0096] In some embodiments of any one of the combinations provided herein, the genetically engineered cells comprise T cells or NK cells. In some embodiments, the genetically engineered cells comprise T cells. In some embodiments, the T cells are CD4+ and/or CD8+. In some embodiments, the T cells are primary T cells obtained from a subject.

[0097] In some embodiments of any one of the combinations provided herein, the genetically engineered cells are formulated as a pharmaceutical composition for administration to a subject, optionally wherein the cells are formulated for administration in one or more unit doses for treating a disease or condition.

[0098] In some embodiments of any one of the combinations provided herein, the gamma secretase inhibitor is formulated as a pharmaceutical composition for administration to a subject, optionally wherein the gamma secretase inhibitor is formulated for administration in one or more unit doses.

[0099] Provided herein are kits that contain: the combination of any of claims 99-139 and instructions for administering the components of the combination to a subject having a disease or disorder.

[0100] Provided herein are kits that contain: (a) a reagent for detecting expression of B cell maturation antigen (BCMA) on the surface of a cell; (b) an inhibitor of gamma secretase, optionally formulated for administration in one or more unit doses; and (c) instructions for administering the inhibitor to a subject based on the results of use of the reagent for detecting BCMA on the surface of plasma cells in a subject having a cancer.

[0101] In some embodiments of any one of the kits provided herein, the instructions specify administering the inhibitor to the subject if the plasma cells comprise low expression of surface BCMA and/or a level of expression of surface BCMA below a threshold level. In some embodiments, the threshold level of expression of surface BCMA is lower than the average or median expression or level of surface BCMA on plasma cells in a plurality of control subjects, optionally wherein the control subjects are a group of healthy or normal subjects. In some embodiments, the low expression of surface BCMA is present when less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma cell marker or phenotype, or the cancer cells in the subject express surface BCMA; or the threshold level of surface BCMA is less than or less than about 60%, less than or less than about 50%, less than or less than about 40%, less than or less than about 30%, less than or less than about 20% or less than or less than about 10% of the plasma cells, or cells with a plasma cell marker or phenotype or the cancer cells in the subject that express surface BCMA.

[0102] In some embodiments of any one of the kits provided herein, the kits further contain: genetically engineered cells expressing a recombinant receptor, optionally wherein the genetically engineered cells are formulated for administration of one or more unit doses to a subject having a disease or condition. In some embodiments, the recombinant receptor specifically binds to a target antigen associated with the disease or condition.

[0103] In some embodiments of any one of the kits provided herein, the instructions specify administering the gamma secretase inhibitor, or one or more unit doses thereof, to a subject having a disease or disorder prior to, concurrently with or after initiation of administration of a dose of the genetically engineered cells to the subject, optionally wherein the genetically engineered cells are formulated for administration of one or more unit doses to a subject having a disease or condition. In some embodiments, the instructions specify administering the gamma secretase inhibitor, or one or more unit doses thereof, to a subject having a disease or disorder prior to initiation of administration of a dose of the genetically engineered cells to the subject. In some embodiments, the instructions specify administering the inhibitor within, or within about, 1 hours, 2 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours or 1 week prior to the initiation of the administration of the cell therapy. In some embodiments, the instructions specify administering the gamma secretase inhibitor, or one or more unit doses thereof, to a subject having a disease or disorder after initiation of administration of a dose of the genetically engineered cells to the subject.

[0104] Provided herein are kits that contain: (a) a gamma secretase inhibitor, optionally wherein the gamma secretase inhibitor is formulated in one or more unit doses; and (b) instructions for administering the gamma secretase inhibitor to a subject after initiation of administration of a cell therapy to a subject, the cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor.

[0105] Provided herein are kits that contain: (a) a gamma secretase inhibitor, optionally wherein the gamma secretase inhibitor is formulated in one or more unit doses; and (b) instructions for administering the gamma secretase inhibitor to a subject after initiation of administration of a cell therapy to a subject, the cell therapy comprising a dose of genetically engineered cells expressing a recombinant receptor comprising a chimeric antigen receptor (CAR) comprising an antigen-binding domain that binds to surface B cell maturation antigen (BCMA), wherein binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay.

[0106] In some embodiments of any one of the kits provided herein, initiation of administration of the inhibitor is within, or within about, 1 hours, 2 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours,1 week, 14 days, 21 days, 28 days or more after the initiation of the administration of the dose of genetically engineered cells.

[0107] In some embodiments of any one of the kits provided herein, the instructions specify the inhibitor is for administeration at a time in which: the number of cells of the cell therapy detectable in the blood from the subject is decreased compared to in the subject at a preceding time point after initiation of the administration of the cells; the number of cells of the cell therapy detectable in the blood is less than or less than about 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 50-fold or 100-fold or less the peak or maximum number of the cells of the cell therapy detectable in the blood of the subject after initiation of administration of the administration of the cells; and/or at a time after a peak or maximum level of the cells of the cell therapy are detectable in the blood of the subject, the number of cells of or derived from the cells detectable in the blood from the subject is less than less than 10%, less than 5%, less than 1% or less than 0.1% of total peripheral blood mononuclear cells (PBMCs) in the blood of the subject.

[0108] In some embodiments of any one of the kits provided herein, the instructions specify the inhibitor is for administration up to 7 days, 14 days, 21 days, 28 days or more after initiation of the administration of the cells.

[0109] In some embodiments of any one of the kits provided herein, the recombinant receptor specifically binds to a target antigen associated with the disease or disorder. In some embodiments, the target antigen is selected from among, B cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, a pathogen-specific antigen and an antigen associated with a universal tag. In some embodiments, the target antigen is Muc 1, optionally human Muc 1. In some embodiments of any embodiment, the target antigen is a human antigen.

[0110] In some embodiments, the target antigen is BCMA, optionally human BCMA. In some embodiments, binding of the CAR to surface BCMA or a measure indicative of function or activity, following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA, optionally at a concentration or amount of the soluble or shed form of the BCMA corresponding to a concentration or amount present in serum or blood or plasma of the subject or of a multiple myeloma patient, or on average in a patient population for the disease or disorder, or at a concentration or amount of the soluble or shed BCMA at which the binding or measure is reduced or blocked, or is substantially reduced or blocked, for cells expressing a reference anti-BCMA CAR in the same assay. In some embodiments, the recombinant receptor does not bind soluble BCMA or binds to soluble BCMA with an affinity lower than the affinity of said recombinant receptor for binding to surface BCMA.

[0111] In some embodiments of any one of the kits provided herein, the recombinant receptor is a transgenic T cell receptor (TCR) or a functional non-T cell receptor. In some embodiments, the recombinant receptor is a chimeric receptor, which optionally is a chimeric antigen receptor (CAR). In some embodiments, the chimeric antigen receptor (CAR) comprises an extracellular antigen-recognition domain that specifically binds to the antigen and an intracellular signaling domain comprising an ITAM. In some embodiments, the intracellular signaling domain comprises and intracellular domain of a CD3-zeta (CD3.zeta.) chain. In some embodiments, the chimeric antigen receptor (CAR) further comprises a costimulatory signaling region. In some embodiments, the costimulatory signaling region comprises a signaling domain derived from CD28 or 4-1BB, optionally human CD28 or human 4-1BB. In some embodiments, the costimulatory signaling region is a domain derived from 4-1BB, optionally human 4-1BB.

[0112] In some embodiments of any one of the kits provided herein, the genetically engineered cells comprise T cells or NK cells. In some embodiments, the genetically engineered cells comprise T cells. In some embodiments, the T cells are CD4+ and/or CD8+. In some embodiments, the T cells are primary T cells obtained from a subject.

[0113] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor inhibits or reduces or is capable of inhibiting or reducing cleavage of one or more targets selected from BCMA, Notch 1, Notch 2, Notch 3, Notch 4, Muc1, Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CXCR1, CXCL16, Delta1, E-cadherein, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R or ameloid precursor protein (APP). In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises Muc 1. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises BCMA.

[0114] In some embodiments of any one of the kits provided herein, cleavage of one or more targets selected from BCMA, Notch 1, Notch 2, Notch 3, Notch 4, Muc1, Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CXCR1, CXCL16, Delta1, E-cadherein, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R or ameloid precursor protein (APP) is inhibited or reduced or can be inhibited or reduced the gamma secretase inhibitor. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises Muc1. In some embodiments, the one or more targets the cleavage of which is reduced or inhibited comprises BCMA.

[0115] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor inhibits the target with a half-maximal inhibitory concentration (IC.sub.50) of 0.01 nM to 1 .mu.M, 0.01 nM to 100 nM, 0.01 nM to 10 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.01 nM to 0.5 nM, 0.01 nM to 0.25 nM, 0.01 nM to 0.1 nM, 0.01 nM to 0.05 nM, 0.05 nM to 1 .mu.M, 0.05 nM to 100 nM, 0.05 nM to 10 nM, 0.05 nM to 5 nM, 0.05 nM to 1 nM, 0.05 nM to 0.5 nM, 0.05 nM to 0.25 nM, 0.05 nM to 0.1 nM, 0.1 nM to 1 .mu.M, 0.1 nM to 100 nM, 0.1 nM to 10 nM, 0.1 nM to 5 nM, 0.1 nM to 1 nM, 0.1 nM to 0.5 nM, 0.1 nM to 0.25 nM, 0.25 nM to 1 .mu.M, 0.25 nM to 100 nM, 0.25 nM to 10 nM, 0.25 nM to 5 nM, 0.25 nM to 1 nM, 0.25 nM to 0.5 nM, 0.5 nM to 1 .mu.M, 0.5 nM to 100 nM, 0.5 nM to 10 nM, 0.5 nM to 5 nM, 0.5 nM to 1 nM, 1 nM to 1 .mu.M, 1 nM to 100 nM, 1 nM to 10 nM, 1 nM to 5 nM, 5 nM to 1 .mu.M, 5 nM to 100 nM, 5 nM to 10 nM, 10 nM to 1 .mu.M, 10 nM to 100 nM or 100 nM to 1 .mu.M.

[0116] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor inhibits the target with a half-maximal inhibitory concentration (IC.sub.50) of less than or less than about 1 .mu.M, 100 nM, 10 nM, 5 nM, 1 nM, 0.5 nM, 0.35 nM, 0.25 nM, 0.1 nM, 0.05 nM or 0.01 nM or less.

[0117] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor is a peptide inhibitor or non-peptide inhibitor. In some embodiments, the gamma secretase inhibitor is a peptide inhibitor and the peptide inhibitor is selected from among peptide aldehydes derivatives, difluoroketones derivatives, hydroxyethylene dipeptide isotere derivatives, alpha-helical peptide derivatives and dipeptide analogs. In some embodiments, the gamma secretase inhibitor is a non-peptide inhibitor and the non-peptide inhibitor is a benzodiazepines derivative or a sulfonamides derivative.

[0118] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor is a transition state inhibitor or non-transition state inhibitor.

[0119] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor is a nonsteroidal anti-inflammatory drug.

[0120] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor is selected from LY3039478, secretase inhibitor I (GSI I) Z-Leu-Leu-Norleucine; .gamma.-secretase inhibitor II (GSI II); .gamma.-secretase inhibitor III (GSI III), N-Benzyloxycarbonyl-Leu-leucinal, N-(2-Naphthoyl)-Val-phenylalaninal; .gamma.-secretase inhibitor III (GSI IV); .gamma.-secretase inhibitor III (GSI V), N-Benzyloxycarbonyl-Leu-phenylalaninal; .gamma.-secretase inhibitor III (GSI VI), 1-(S)-endo-N-(1,3,3)-Trimethylbicyclo[2.2.1]hept-2-yl)-4-fluorophenyl Sulfonamide; .gamma.-secretase inhibitor III (GSI VII), Menthyloxycarbonyl-LL-CHO; .gamma.-secretase inhibitor III (GSI IX), (DAPT), N-[N-(3,5-Difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester; .gamma.-secretase inhibitor X (GSI X), {1S-Benzyl-4R-[1-(1S-carbamoyl-2-phenethylcarbamoyl)-1S-3-methylbutylcarb- -amoyl]-2R-hydroxy-5-phenylpentyl}carbamic Acid tert-butyl Ester; .gamma.-secretase inhibitor XI (GSI XI), 7-Amino-4-chloro-3-methoxyisocoumarin; .gamma.-secretase inhibitor XII (GSI XII), Z-Ile-Leu-CHO; .gamma.-secretase inhibitor XIII (GSI XIII), Z-Tyr-Ile-Leu-CHO; .gamma.-secretase inhibitor XIV (GSI XIV), Z-Cys(t-Bu)-Ile-Leu-CHO; .gamma.-secretase inhibitor XVI (GSI XVI), N-[N-3,5-Difluorophenacetyl]-L-alanyl-S-phenylglycine Methyl Ester; .gamma.-secretase inhibitor XVII (GSI XVII); .gamma.-secretase inhibitor XIX (GSI XIX), benzo[e][1,4]diazepin-3-yl)-butyramide; .gamma.-secretase inhibitor XX (GSI XX), (S,S)-2-[2-(3,5-Difluorophenyl)acetylamino]-N-(5-methyl-6-oxo-6,7-dihydro- -5H-dibenzo[b,d]azepin-7-yl)propionamide; .gamma.-secretase inhibitor XXI (GSI XXI), (S,S)-2-[2-(3,5-Difluorophenyl)-acetylamino]-N-(1-methyl-2-oxo-5-phenyl-2- -,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-propionamide; Gamma40 secretase inhibitor I, N-trans-3,5-Dimethoxycinnamoyl-Ile-leucinal; Gamma40 secretase inhibitor II, N-tert-Butyloxycarbonyl-Gly-Val-Valinal Isovaleryl-V V-Sta-A-Sta-OCH3; MK-0752; MRK-003 (Merck); semagacestat/LY450139; RO4929097; PF-03084,014; BMS-708163; MPC-7869 (.gamma.-secretase modifier), YO-01027 (Dibenzazepine), Compound E ([(2S)-2-{[(3,5-Difluorophenyl)acetyl]amino}-N-[(3S)-1-methyl-2-oxo-5-phe- nyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide], LY411575, L-685,458, BMS-289948 (4-chloro-N-(2,5-difluorophenyl)-N-((1R)-{4-fluoro-2-[3-(1H-imidazol-1-yl- )propyl]phenyl}ethyl)benzenesulfonamide hydrochloride) and BMS-299897 (4-[2-((1R)-1-{[(4-chlorophenyl)sulfonyl]-2,5-difluoroanilino}ethyl)-5-fl- uorophenyljbutanoic acid).

[0121] In some embodiments of any one of the kits provided herein, the gamma secretase inhibitor is a compound of the structure:

##STR00008##

[0122] or a pharmaceutically acceptable salt of hydrate thereof.

[0123] In some embodiments of any one of the kits provided herein, the instructions specify administering a dose of genetically engineered cells to a subject having a disease or disorder. In some embodiments, the disease or disorder is a cancer. In some embodiments, the cancer is a B cell malignancy. In some embodiments, the cancer is multiple myeloma, plasmacytoma, a cancer of plasma cell origin and/or a cancer of B cell origin.

[0124] In some embodiments of any one of the kits provided herein, the dose comprises from or from about 1.times.10.sup.5 to 5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs); the cell therapy comprises the administration of from or from about 1.times.10.sup.5 to 1.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total PBMCs; from or from about 5.times.10.sup.5 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs; or from or from about 1.times.10.sup.6 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs, each inclusive; optionally wherein the instructions specify the administration of one or of a plurality of unit doses comprising the dose of cells and/or a volume corresponding to such one or plurality of unit doses comprising the dose of cells.

[0125] In some embodiments of any one of the kits provided herein, the dose comprises no more than 5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs); no more than 2.5.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 1.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 0.5.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 1.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total PBMCs; no more than 0.5.times.10.sup.6 total recombinant receptor-expressing cells, total T cells, or total PBMCs; optionally wherein the instructions specify the administration of one or of a plurality of unit doses comprising the dose of cells and/or a volume corresponding to such one or plurality of unit doses comprising the dose of cells.

[0126] In some embodiments of any one of the kits provided herein, the instructions specify administering the inhibitor orally, subcutaneously or intravenously. In some embodiments, the instructions specify administering the inhibitor orally.

[0127] In some embodiments of any one of the kits provided herein, the instructions specify the inhibitor is to be administered at least six times daily, five times daily, four times daily, three times daily, twice daily, once daily, every other day, three times a week, at least once a week, or only one time. In some embodiments, the instructions specify the inhibitor is administered three times a week.

[0128] In some embodiments of any one of the kits provided herein, the instructions specify the administration of the inhibitor is to be carried out in a treatment cycle that is at least or at least about or 14 days, at least or at least about or 21 days or at least or at least about or 28 days.

[0129] In some embodiments of any one of the kits provided herein, the instructions specify the inhibitor is administered, or each administration of the inhibitor is independently administered, in an amount of 0.5 mg to 500 mg, 0.5 mg to 250 mg, 0.5 mg to 100 mg, 0.5 mg to 50 mg, 0.5 mg to 25 mg, 0.5 mg to 10 mg, 0.5 mg to 5.0 mg, 0.5 mg to 2.5 mg, 0.5 mg to 1.0 mg, 1.0 mg to 500 mg, 1.0 mg to 250 mg, 1.0 mg to 100 mg, 1.0 mg to 50 mg, 1.0 mg to 25 mg, 1.0 mg to 10 mg, 1.0 mg to 5.0 mg, 1.0 mg to 2.5 mg, 2.5 mg to 500 mg, 2.5 mg to 250 mg, 2.5 mg to 100 mg, 2.5 mg to 50 mg, 2.5 mg to 25 mg, 2.5 mg to 10 mg, 2.5 mg to 5.0 mg, 5.0 mg to 500 mg, 5.0 mg to 250 mg, 5.0 mg to 100 mg, 5.0 mg to 50 mg, 5.0 mg to 25 mg, 5.0 mg to 10 mg, 10 mg to 500 mg, 10 mg to 250 mg, 10 mg to 100 mg, 10 mg to 50 mg, 10 mg to 25 mg, 25 mg to 500 mg, 25 mg to 250 mg, 25 mg to 100 mg, 25 mg to 50 mg, 50 mg to 500 mg, 50 mg to 250 mg, 50 mg to 100 mg, 100 mg to 500 mg, 100 mg to 250 mg or 250 mg to 500 mg.

[0130] In some embodiments of any one of the kits provided herein, the instructions specify the inhibitor is administered, or each administration of the inhibitor is independently administered, in an amount that is at least or at least about or is or is about 0.5 mg, 1.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 25 mg, 50 mg, 100 mg, 250 mg or 500 mg.

BRIEF DESCRIPTION OF THE DRAWINGS

[0131] FIG. 1 shows expression level of surface B Cell Maturation Antigen (BCMA) on OPM2, MM1s or RPMI-8826 cells co-cultured with different concentrations of a representative gamma secretase inhibitor (LY3039478) after 24 hours of co-culture. The bar near the X-axis suggests the estimated C.sub.max and C.sub.min for the inhibitor.

[0132] FIG. 2A and FIG. 2B show percent lysis (% killing) of OPM2 cells (FIG. 2A) or RPMI-8226 cells (FIG. 2B) co-cultured with anti-BCMA CAR-expressing T cells in the presence or absence of a representative of gamma secretase inhibitor (LY3039478) after about 150 hours of co-culture. FIG. 2C shows percent lysis (% killing) of OPM2 cells co-cultured with anti-BCMA CAR-expressing T cells in the presence or absence of a representative gamma secretase inhibitor (LY3039478). The bar near the X-axis suggests the estimated C.sub.max and C.sub.min for the inhibitor.

[0133] FIGS. 3A-3D show the production of IFN-gamma and IL-2 in the supernatant of anti-BCMA CAR-expressing T cells co-cultured with OPM2 (FIGS. 3A and 3C) or RPMI-8226 cells (FIGS. 3B and 3D) in the presence or absence of a representative of gamma secretase inhibitor (LY3039478) after 24 hours of co-culture. FIG. 3E shows the production of IFN-gamma in the supernatant of anti-BCMA CAR-expressing T cells co-cultured with OPM2 cells and different concentrations of a representative gamma secretase inhibitor (LY3039478). The bar near the X-axis suggests the estimated C.sub.max and C.sub.min for the inhibitor.

[0134] FIGS. 4A and 4B show the cell number of anti-BCMA CAR-expressing T cells originally obtained from a healthy donor (FIG. 4A) and a donor with multiple myeloma (FIG. 4B) after multiple rounds of stimulation with MM1S cells over a period of 10-17 days in the absence of a representative of gamma secretase inhibitor (LY3039478) (vehicle control, square) or in the presence of the inhibitor with a concentration of 1 .mu.M (upward triangle) or 0.001 .mu.M (downward triangle).

[0135] FIGS. 5A and 5B show the production of IFN-gamma, IL2, and TNF-alpha in the supernatant of anti-BCMA CAR-expressing T cells originally obtained from a healthy donor (FIG. 5A) and a donor with multiple myeloma (FIG. 5B) after a second round of stimulation with MM1S cells at day 4 a in the absence of a representative of gamma secretase inhibitor (LY3039478) (left bar) or in the presence of the inhibitor with a concentration of 0.001 .mu.M (middle bar) or 0.001 .mu.M (right bar).

[0136] FIG. 6 shows plasma levels of gamma secretase inhibitor LY3039478 at different time points following oral administration of a single dose of LY3039478 (3 mg/kg).

[0137] FIG. 7 shows plasma levels of BCMA over time following oral administration of a single dose of LY3039478 (3 mg/kg) in a mouse xenograft model for human multiple myeloma (RPMI-8226).

[0138] FIG. 8 shows surface BCMA expression, as assessed by flow cytometry, in subcutaneous tumor cells derived from a mouse xenograft model for human multiple myeloma (RPMI-8226) over time following oral administration of a single dose of LY3039478.

[0139] FIGS. 9A-9B show results from the study described in Example 5, assessing the impact of different treatments in a mouse xenograft model for human multiple myeloma. FIG. 9A depicts tumor volume (mm.sup.3) over days during the study. The dashed arrow below the graph indicates the time period during which LY3039478 was administered as described in Example 5. FIG. 9B shows percent survival among animals treated over the course of 65 days following CAR T injection.

[0140] FIGS. 10A-10B show CD4+ CAR T+ and CD8+ CAR T+ peripheral blood counts over time (days) following CAR T injection (Days Post-CAR T injection) of animals of a human multiple myeloma xenograft mouse model treated under various conditions as described in Example 5. FIG. 10A depicts CD4+ CAR T+ cell counts per .mu.L of blood over time. FIG. 10B depicts CD8+ CAR T+ cell counts per .mu.L of blood over time GSI bar with arrows indicates the timing of LY3039478 (GSI) delivery if administered.

[0141] FIG. 11 shows CD4+ (top) and CD8+ (bottom) CAR+ T cell counts per 1.times.10.sup.6 cells from satellite tumor digests taken on days 7 and 15 following low dose (1e6) CAR T injection in the study described in Example 5.

[0142] FIG. 12A shows serum levels of BCMA on days 7 and 15 following commencement of different treatment regimes as describe in Example 5.

[0143] FIG. 12B shows surface BCMA expression, as assessed by flow cytometry, from tumor digests taken on days 7 and 15 following anti-BCMA CAR T cell injection in mice from various treatment groups as described in Example 5.

[0144] FIG. 13A shows RPMI-8226, OPM2, and MM1.S cell viability over time when treated with chronically stimulated anti-BCMA CAR+ T cells and DMSO (DMSO control) or the gamma secretase inhibitor compound LY3039478 (1 .mu.M GSI), at an effector to target ratio of 0.3:1, or when untreated (RPMI, OPM2, or MM1.S alone).

[0145] FIG. 13B shows the percentage of killing of RPMI-8226, OPM2, and MM1.S cells when treated with chronically stimulated anti-BCMA CAR+ T cells and DMSO (DMSO control) or LY3039478 (1 .mu.M GSI), or when untreated (RPMI, OPM2, or MM1.S alone).

[0146] FIG. 13C shows supernatant IL-2 concentrations following 24 hour incubation of RPMI-8226, OPM2, and MM1.S cells with chronically stimulated anti-BCMA CAR+ T cells and LY3039478 (1 .mu.M GSI) or DMSO (DMSO control) at effector to target ratios of 0.3:1 or 1:1.

[0147] FIG. 14 shows IFNg, IL-2, and TNFa concentrations in supernatant following 24 hour incubation of anti-BCMA CAR+ T cells with BCMA-containing beads in different concentrations of gamma secretase inhibitor LY3039478 or DMSO (veh.).

[0148] FIG. 15A shows BCMA antibody binding capacity (BCMA ABC) on the surface of cells in samples derived from multiple myeloma patients cells before (Before GSI) and after (After GSI) administration of three doses of the small molecule gamma secretase inhibitor (GSI) LY3039478 by mouth as described in Example 8.

[0149] FIG. 15B shows the percentage of plasma cells in patient samples determined to have measurable surface BCMA expression, before (Before GSI) and after (After GSI) administration of three doses of the small molecule gamma secretase inhibitor (GSI) LY3039478 by mouth, as described in Example 8.

DETAILED DESCRIPTION

[0150] Provided herein are combination therapies involving administration of an immunotherapy, such as a cell therapy, e.g a T cell therapy, and an inhibitor of gamma secretase. In some aspects, the provided methods involve administration of an immunotherapy or immunotherapeutic agent, such as a composition including cells for adoptive cell therapy, e.g., such as a T cell therapy (e.g. CAR-expressing T cells) and administration of a gamma secretase inhibitor to a subject having a disease or disorder. In some embodiments, the gamma secretase is administered prior to, concurrently with or subsequent to initiation of administration of the immunotherapy, e.g. cell therapy. In some embodiments, the gamma secretase inhibitor inhibits or reduces intramembrane cleavage of a target of a gamma secretase, e.g. BCMA, on a cell (such as a tumor/cancer cell). In some aspects, the target is the same as the target antigen of the cell therapy. In some embodiments, the cell therapy, such as CAR-expressing T cells, comprises an antigen-binding domain that binds to a B Cell Maturation Antigen (BCMA), such as surface BCMA. In some embodiments, the methods further comprise selecting a subject for treatment, wherein the subject has a low expression of surface BCMA on a cell, such as a tumor/cancer cell, e.g. cells of the cancer in the subject that express CD138, surface CD38 or a surface plasma cell marker or are derived from plasma cells. In some embodiments, the cell therapy comprises a recombinant receptor, such as chimeric antigen receptor (CAR), that binds to an antigen other than BCMA. In some embodiments, the methods are carried out in connection with adoptive cell therapy.

[0151] Cell therapies, such as T cell-based therapies, for example, adoptive T cell therapies (including those involving the administration of cells expressing chimeric receptors specific for a disease or disorder of interest, such as chimeric antigen receptors (CARs) and/or other recombinant antigen receptors, as well as other adoptive immune cell and adoptive T cell therapies) can be effective in the treatment of cancer and other diseases and disorders. The engineered expression of recombinant receptors, such as chimeric antigen receptors (CARs), on the surface of T cells enables the redirection of T-cell specificity. In clinical studies, CAR-T cells, for example anti-CD19 CAR-T cells, have produced durable, complete responses in both leukemia and lymphoma patients (Porter et al. (2015) Sci Transl Med., 7:303ra139; Kochenderfer (2015) J. Clin. Oncol., 33: 540-9; Lee et al. (2015) Lancet, 385:517-28; Maude et al. (2014) N Engl J Med, 371:1507-17).

[0152] In certain contexts, available approaches to adoptive cell therapy may not always be entirely satisfactory. In some contexts, optimal efficacy can depend on the ability of the administered cells to recognize and bind to a target, e.g., target antigen, to traffic, localize to and successfully enter appropriate sites within the subject, tumors, and environments thereof. In some contexts, optimal efficacy can depend on the ability of the administered cells to become activated, expand, to exert various effector functions, including cytotoxic killing and secretion of various factors such as cytokines, to persist, including long-term, to differentiate, transition or engage in reprogramming into certain phenotypic states (such as long-lived memory, less-differentiated, and effector states), to avoid or reduce immunosuppressive conditions in the local microenvironment of a disease, to provide effective and robust recall responses following clearance and re-exposure to target ligand or antigen, and avoid or reduce exhaustion, anergy, peripheral tolerance, terminal differentiation, and/or differentiation into a suppressive state.

[0153] In some embodiments, the exposure and persistence of engineered cells is reduced or declines after administration to the subject. Yet, observations indicate that, in some cases, increased exposure of the subject to administered cells expressing the recombinant receptors (e.g., increased number of cells or duration over time) may improve efficacy and therapeutic outcomes in adoptive cell therapy. Preliminary analyses conducted following the administration of different CD19-targeting CAR-expressing T cells to subjects with various CD19-expressing cancers in multiple clinical trials revealed a correlation between greater and/or longer degree of exposure to the CAR-expressing cells and treatment outcomes. Such outcomes included patient survival and remission, even in individuals with severe or significant tumor burden.

[0154] In some aspects, the provided methods and uses provide for or achieve improved or more durable responses or efficacy as compared to certain alternative methods, such as in particular groups of subjects treated. In some embodiments, the methods are advantageous by virtue of administering T cell therapy, such as a composition including cells for adoptive cell therapy, e.g., such as a T cell therapy (e.g. CAR-expressing T cells), and an inhibitor of gamma secretase.

[0155] The provided methods are based on observations that gamma secretase inhibitor improves one or more functional activities of recombinant receptor-expressing cells specific to BCMA following exposure to BCMA-expressing cells in the presence of a gamma secretase inhibitor. Such functional activities include, for example, antigen-dependent cytolytic activity and/or ability to produce one or more cytokines. In some aspects, the improved activity may be due to increased expression or stabilized expression of surface BCMA on cells (e.g., tumor/cancer cells, for example, multiple myeloma cells). In some aspects, increasing or stabilizing the expression of a target antigen, e.g. BCMA, on target cells improves the outcome for patients who may be or are low for expression of the target antigen, e.g. BCMA, so that sufficient antigen is available for targeting by the cell therapy, e.g. T cell therapy. In some embodiments, the provided embodiments reduce the variability in treatment outcomes among a group of subjects with variable surface expression of the target antigen, e.g. BCMA. In some embodiments of the provided methods, a patient is selected for administration of the provided combination therapy of a gamma secretase inhibitor and an immunotherapy, such as cell therapy (e.g. CAR-T cells) if cells of a cancer in the subject, such as cells that express CD138, surface CD38 or a surface plasma cell marker or are derived from plasma cells, have low expression of surface B cell maturation antigen (BCMA) and/or a level of surface BCMA about a threshold level. Hence, in some aspects, the gamma secretase inhibitor is administered in an amount to stabilize surface expression of BCMA, such as by reducing or inhibiting intramembrane cleavage of BCMA.

[0156] In some embodiments, the provided methods offer advantages in subjects that do not have, or that are not selected based on, high levels of serum or plasma or tumor levels of soluble or shed BCMA. In some embodiments, the patient is not selected for having high serum or plasma or tumor levels of soluble or shed BCMA. In some embodiments, the provided combination therapy involves administration of a cell therapy, e.g. CAR-expressing T cells, having an antigen-binding domain that binds to surface BCMA in which the antigen-binding domain binds to soluble BCMA with an affinity lower than the affinity of said antigen-binding domain binding to surface BCMA. In some contexts, the provided combination therapy involves administration of a cell therapy, such as recombinant receptor-expressing cells, e.g. CAR-expressing T cells, in which binding to surface BCMA by the recombinant receptor or CAR or a measure indicative of function or activity of the recombinant receptor-expressing cells (e.g. CAR-expressing cells), following exposure to cells expressing surface BCMA, is not reduced or blocked or is not substantially reduced or blocked in the presence of a soluble or shed form of the BCMA.

[0157] In some aspects, such results are achieved while enhancing or improving the activity, in some cases the intrinsic activity, of cells, e.g. T cells, of the cell therapy. In some aspects, this can be achieved in the presence of a gamma secretase inhibitor that exhibits activity for other targets beyond BCMA, such as activity for cleavage of a Notch. An example of such an inhibitor is LY3039478 or compound 1 herein, or stereoisomers, pharmaceutically acceptable salts or hydrates thereof. In some aspects, although notch signaling can regulate T cell differentiation and/or proliferation, observations herein demonstrate enhanced activities of T cells in the presence of an exemplary inhibitor known to target notch signaling, e.g. LY3039478. For example, as shown herein, in an exemplary serial stimulation assay assessing anti-BCMA CAR-T cell activities following repeated stimulation with BCMA-expressing cells, increased CAR+ T cell expansion and/or survival was observed in the presence of the exemplary gamma secretase inhibitor. Such results indicate that the provided combination therapy improves T cell function, including functions related to the expansion, proliferation and persistence of T cells.

[0158] The provided findings indicate that combination therapy of the gamma secretase inhibitor in methods involving cell therapy, such as involving administration of adoptive T cell therapy, achieves improved function of the T cell therapy. In some embodiments, combination of the cell therapy (e.g., administration of engineered T cells) with the gamma secretase inhibitor improves or enhances one or more functions and/or effects of the T cell therapy, including after encounter with antigen, such as persistence, expansion, cytotoxicity, and/or therapeutic outcomes, e.g., ability to kill or reduce the burden of tumor or other disease or target cell. In some aspects, the provided methods increase overall response and/or survival by or more than 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold or more compared to an alternative treatment, such as compared to a monotherapy involving administration of the T cell therapy (e.g. CAR-T cell) or a gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof.

[0159] In some aspects, the provided methods can enhance, increase or potentiate T cell therapy, such as to overcome lack of persistence and/or exhaustion of T cells. In some embodiments, a subject having received administration of a T cell therapy, e.g. CAR-T cell, is monitored for the presence, absence or level of T cells of the therapy in the subject, such as in a biological sample of the subject, e.g. in the blood of the subject. In some embodiments, a gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, is administered to a subject having received the T cell therapy (e.g. CAR-T cells) but in which such cells have weakly expanded and/or are at or below a threshold level in a sample of the subject, e.g. blood sample, at a time when strong or robust expansion of the CAR-T cells in the subject is typically observed in a plurality of subjects administered a T cell therapy (e.g. CAR-T), in some cases, this same T cell therapy (e.g. same CAR-T cells). In some of any such embodiments, the level of engineered, e.g., CAR+, cells in the sample is determined as the number of the cells, e.g., CAR+ cells, per microliter of the sample; in some embodiments, the peak level is the highest such measurement following, optionally over a specified period of time following, administration of the cells or cell therapy to the subject. In some aspects, a gamma secretase inhibitor is administered to a subject having previously received a cell therapy, if, at or about days 1-15 days after initiation of administration of the T cell therapy to the subject, less than 10 cells per .mu.L, such as less than 5 cells per .mu.L or less than 1 cells per .mu.L of cells of the T cell therapy, e.g. CAR-T cells, or a CD8+ or CD3+ subset thereof, are detectable in the blood. In some embodiments, a gamma secretase inhibitor is administered to a subject having previously received a cell therapy, if, at or about day 12-15 cells after initiation of administration of the T cell therapy to the subject, e.g. CAR-T cells, less than 10 cells per .mu.L, such as less than 5 cell pers .mu.L or less than 1 cell per .mu.L of such cells, or a CD8+ or CD3+ subset thereof, are detectable in the blood.

[0160] In certain aspects, the provided methods can enhance, increase or potentiate T cell therapy in subjects in which a peak response (e.g. presence of T cells and/or reduction in tumor burden) to the T cell therapy has been observed, but in which the response, e.g. presence of T cells and/or reduction in tumor burden, has become reduced or is no longer detectable. In some aspects, a gamma secretase inhibitor is administered to a subject within a week, such as within 1, 2 or 3 days after: (i) peak or maximum level of the cells of the T cell therapy are detectable in the blood of the subject; (ii) the number of cells of the T cell therapy detectable in the blood, after having been detectable in the blood, is not detectable or is reduced, optionally reduced compared to a preceding time point after administration of the T cell therapy; (iii) the number of cells of the T cell therapy detectable in the blood is decreased by or more than 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold or more compared to the peak or maximum number cells of the T cell therapy detectable in the blood of the subject after initiation of administration of the T cell therapy; (iv) at a time after a peak or maximum level of the cells of the T cell therapy are detectable in the blood of the subject, the number of cells of or derived from the T cells detectable in the blood from the subject is less than 10%, less than 5%, less than 1% or less than 0.1% of total peripheral blood mononuclear cells (PBMCs) in the blood of the subject; (v) the subject exhibits disease progression and/or has relapsed following remission after treatment with the T cell therapy; and/or (iv) the subject exhibits increased tumor burden as compared to tumor burden at a time prior to or after administration of the T cells and prior to initiation of administration of the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof.

[0161] In some embodiments, a gamma secretase inhibitor is administered to a subject having received a cell therapy, e.g. CAR-T cells, but who has relapsed following treatment with the T cell therapy, such as at a time in which the response, e.g. presence of T cells and/or reduction in tumor burden, has become reduced or is no longer detectable. In some aspects, relapse may occur due to BCMA antigen loss, which, in some cases, can be due to BCMA antigen downregulation/antigen escape. In some cases, it may be due to cleavage or shedding of BCMA antigen from the cells surface. In some aspects, antigen loss may lead to a reduction in cells presenting a target antigen (e.g., BCMA), thereby diminishing or reducing the activity and/or function of the CAR T cells and/or decreasing the number of CAR T cells, for example, in the blood. In some embodiments, administration of gamma secretase inhibitor can be administered at a time in which the subject has relapsed or is suspected or likely to relapse to cell therapy to reduce or prevent cleavage or shedding of the target antigen (e.g., BCMA) from the cell surface. In such embodiments, administering gamma secretase inhibitor can reinvigorate or boost CAR T cells following relapse by preventing or reducing cleavage and/or shedding of the target antigen (e.g., BCMA).

[0162] In some embodiments, the provided methods involving combination therapy with a cell therapy and a gamma secretase inhibitor result in genetically engineered cells with increased persistence and/or better potency in a subject to which it is administered compared to administration of the cell therapy in the absence of the gamma secretase inhibitor. In some embodiments, the persistence is increased at least or about at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold or more. In some embodiments, the degree or extent of persistence of administered cells can be detected or quantified after administration to a subject. For example, in some aspects, quantitative PCR (qPCR) is used to assess the quantity of cells expressing the recombinant receptor (e.g., CAR-expressing cells) in the blood or serum or organ or tissue (e.g., disease site) of the subject. In some aspects, persistence is quantified as copies of DNA or plasmid encoding the receptor, e.g., CAR, per microgram of DNA, or as the number of receptor-expressing, e.g., CAR-expressing, cells per microliter of the sample, e.g., of blood or serum, or per total number of peripheral blood mononuclear cells (PBMCs) or white blood cells or T cells per microliter of the sample. In some embodiments, flow cytometric assays detecting cells expressing the receptor generally using antibodies specific for the receptors also can be performed. Cell-based assays may also be used to detect the number or percentage of functional cells, such as cells capable of binding to and/or neutralizing and/or inducing responses, e.g., cytotoxic responses, against cells of the disease or condition or expressing the antigen recognized by the receptor. In any of such embodiments, the extent or level of expression of another marker associated with the recombinant receptor (e.g. CAR-expressing cells) can be used to distinguish the administered cells from endogenous cells in a subject.

[0163] In aspects of the methods provided herein, combining cell therapy with a gamma secretase inhibitor that increases potentcy and/or promotes sustained CAR T cell activity allows administration of a lower dose of CAR T cells to achieve the same therapeutic effect as that seen with a higher dose of CAR T cells without a gamma secretase inhibitor. In some embodiments, the dose of CAR T cells administered when combined with gamma secretase inhibitor can be reduced by 10%, 20%, 30%, 40%, 50%, 60%, 70%, or more of a dose of CAR T cells administered without gamma secretase inhibitor.

[0164] In some embodiments, the methods can be used for treating a disease or condition, such as a cancer, including a B cell malignancy or hematological malignancy. In some aspects, the cancer is multiple myeloma, plasmacytoma, a cancer of plasma cell origin and/or a cancer of B cell origin. In some aspects, such diseases, conditions or malignancies include those in which responses, e.g. complete response (CR), to treatment with the cell therapy alone, such as a composition including cells for adoptive cell therapy, e.g., such as a T cell therapy (e.g. CAR-expressing T cells), is relatively low compared to treatment with other T cell therapies or treatment of other diseases or malignacies (e.g. a CR in less than or less than about 60%, less than about 50% or less than about 45% of the subjects so treated).

[0165] In some embodiments, the provided methods reduce or ameliorate a symptom or outcome or burden of the disease or condition to a degree that is greater than the combination of (i) the degree of reduction or amelioration effected by the administration of the gamma secretase inhibitor alone, optionally on average in a population of subjects having the disease or condition, and (ii) the degree of reduction or amelioration by the administration of the T cell therapy alone, optionally on average in a population of subjects having the disease or condition. In some embodiments, the method reduces or ameliorates such symptoms, outcomes or burdens of the disease, e.g. compared to on average in a population of subjects having the disease or condition, by greater than or greater than about 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 6.0-fold, 7.0-fold, 8.0-fold, 9.0-fold, 10.0 fold, 20.0-fold, 30.0-fold, 40.0-fold, 50.0-fold or more.

[0166] Also provided are methods for engineering, preparing, and producing the cells (e.g. CAR-expressing T cells), compositions containing the cells and/or gamma secretase inhibitor, and kits and devices containing and for using, producing and administering the cells and/or gamma secretase inhibitor, such as in accord with the provided combination therapy methods.

[0167] All publications, including patent documents, scientific articles and databases, referred to in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication were individually incorporated by reference. If a definition set forth herein is contrary to or otherwise inconsistent with a definition set forth in the patents, applications, published applications and other publications that are herein incorporated by reference, the definition set forth herein prevails over the definition that is incorporated herein by reference.

[0168] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

I. COMBINATION THERAPY

[0169] Provided herein are methods for combination therapy for treating a disease or disorder, e.g. a cancer or proliferative disease, that include administering to a subject a combination therapy of 1) a gamma secretase inhibitor and 2) an immunotherapy, e.g. a cell therapy, e.g. T cell therapy, e.g., CAR-expressing cell (e.g., T cell). In some embodiments, the cell therapy is an adoptive immune cell therapy comprising T cells that specifically recognize and/or target an antigen associated with a disease or disorder, e.g. a cancer or proliferative disease. Also provided are combinations and articles of manufacture, such as kits, that contain a composition comprising the immunotherapy, e.g. T cell therapy, and/or a composition comprising the gamma secretase inhibitor, and uses of such compositions and combinations to treat or prevent diseases, conditions, and disorders, including cancers.

[0170] In some embodiments, such methods can include administration of the gamma secretase inhibitor prior to, simultaneously with, during, during the course of (including once and/or periodically during the course of), and/or subsequently to, the administration (e.g., initiation of administration) of the immunotherapy, e.g. cell therapy (e.g. CAR-expressing T cells). In some embodiments, the administrations can involve sequential or intermittent administrations of the gamma secretase inhibitor and cell therapy.

[0171] In some embodiments, the cell therapy is adoptive cell therapy. In some embodiments, the cell therapy is or comprises a tumor infiltrating lymphocytic (TIL) therapy, a transgenic TCR therapy or a recombinant-receptor expressing cell therapy (optionally T cell therapy), which optionally is a chimeric antigen receptor (CAR)-expressing cell therapy. In some embodiments, the therapy is a B cell targeted therapy. In some embodiments, the therapy targets B cell maturation antigen (BCMA). In some embodiments, the therapy targets CD19. In some embodiments, the therapy targets cell surface associated Mucin 1 (MUC1). In some embodiments, the cells and dosage regimens for administering the cells can include any as described in the following subsection A under "Administration of Cells." In some embodiments, the dosage regimens for administering the gamma secretase inhibitor can include any as described in the following subsection B under "Administration of gamma secretase inhibitor."

[0172] In some embodiments, the immunotherapy, e.g. cell therapy (e.g. CAR-expressing T cells), and gamma secretase inhibitor are provided as pharmaceutical compositions for administration to the subject. In some embodiments, the pharmaceutical compositions contain therapeutically effective amounts of one or both of the agents for combination therapy, e.g., T cells for adoptive cell therapy and a gamma secretase inhibitor as described. In some embodiments, the agents are formulated for administration in separate pharmaceutical compositions. In some embodiments, any of the pharmaceutical compositions provided herein can be formulated in dosage forms appropriate for each route of administration.

[0173] In some embodiments, the combination therapy, which includes administering an immunotherapy, e.g. cell therapy, including engineered cells, such as CAR-T cell therapy, and the gamma secretase inhibitor is administered to a subject or patient having a disease or condition to be treated (e.g. cancer) or at risk for having the disease or condition (e.g. cancer). In some aspects, the methods treat, e.g., ameliorate one or more symptom of, the disease or condition, such as by lessening tumor burden in a cancer expressing an antigen recognized by the immunotherapy or immunotherapeutic agent, e.g. recognized by an engineered T cell.

[0174] In some embodiments, the disease or condition that is treated can be any in which expression of an antigen is associated with and/or involved in the etiology of a disease condition or disorder, e.g. causes, exacerbates or otherwise is involved in such disease, condition, or disorder. Exemplary diseases and conditions can include diseases or conditions associated with malignancy or transformation of cells (e.g. cancer), autoimmune or inflammatory disease, or an infectious disease, e.g. caused by bacterial, viral or other pathogens. Exemplary antigens, which include antigens associated with various diseases and conditions that can be treated, include any of antigens described herein. In particular embodiments, the recombinant receptor expressed on engineered cells of a combination therapy, including a chimeric antigen receptor or transgenic TCR, specifically binds to an antigen associated with the disease or condition.

[0175] In some embodiments, the disease or condition is a cancer or proliferative disease that expresses BCMA. In some embodiments, the provided methods employ a recombinant receptor-expressing cell (e.g. CAR-T cell) that targets BCMA. In particular embodiments, the disease or condition is a multiple myeloma. In some cases, the multiple myeloma is a relapsed or refractory multiple myeloma.

[0176] Among the diseases to be treated is any disease or disorder associated with BCMA or any disease or disorder in which BCMA is specifically expressed and/or in which BCMA has been targeted for treatment (also referred to herein interchangeably as a "BCMA-associated disease or disorder"). Cancers associated with BCMA expression include hematologic malignancies such as multiple myeloma, Waldenstrom macroglobulinemia, as well as both Hodgkin's and non-Hodgkin's lymphomas. See Coquery et al., Crit Rev Immunol., 2012, 32(4):287-305 for a review of BCMA. Since BCMA has been implicated in mediating tumor cell survival, it is a potential target for cancer therapy. Chimeric antigen receptors containing anti-BCMA antibodies, including mouse anti-human BCMA antibodies and human anti-human antibodies, and cells expressing such chimeric receptors have been previously described. See Carpenter et al., Clin Cancer Res., 2013, 19(8):2048-2060, WO 2016/090320, WO2016090327, WO2010104949A2 and WO2017173256. Exemplary CARS containing anti-BCMA antibodies are described herein.

[0177] In some embodiments, prior to the initiation of administration of the engineered cells, such as anti-BCMA CAR, the subject has received one or more prior therapies. In some embodiments, the subject has received at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 or more prior therapies. In some embodiments, the subject has received at least 3, 4, 5, 6, 7, 8, 9, 10 or more prior therapies. In some aspects, the subject has relapsed following, or has been treatment refractory to, one or more of, for example, each, individually, of the one or more prior therapies. In some aspects, the prior therapies include treatment with autologous stem cell transplant (ASCT); an immunomodulatory agent, e.g. an IMiD; a proteasome inhibitor; and an anti-CD38 antibody; unless the subject was not a candidate for or was contraindicated for one or more of the therapies. In some embodiments, the immunomodulatory agent, e.g. an IMiD, is selected from among thalidomide, lenalidomide or pomalidomide. In some embodiments, the proteasome inhibitor is selected from among bortezomib, carfilzomib or ixazomib. In some embodiments, the anti-CD38 antibody is or comprises daratumumab. In some embodiments, the subject must have undergone at least 2 consecutive cycles of treatment for each regimen unless progressive disease was the best response to the regimen. In some aspects, a subject for treatment with the method herein has a myeloma that has relapsed or is treatment refractory with greater than 10% CD138+ malignant plasma cells immunohistochemistry (IHC) on bone marrow core biopsy, either following ASCT or, if the subject has not undergone ASCT, the subject is transplant ineligible, e.g. due to age comorbidity, patient choice, state of disease and/or discretion of physician, and has disease that has persisted after more than four cycles of induction therapy and/or that is refractory to or is not tolerant to therapy with a proteasome inhibitor and immunomodulatory drug, e.g. IMiD.

[0178] In some embodiments, the disease or condition is a tumor, such as a solid tumor, lymphoma, leukemia, blood tumor, metastatic tumor, or other cancer or tumor type.

[0179] In some embodiments, the antigen associated with the disease or disorder is selected from the group consisting of B cell maturation antigen (BCMA), ROR1, Her2, L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, 3, or 4, erbB dimers, EGFR vIII, FBP, FCRL5, FCRH5, fetal acethycholine e receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kdr, kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, EGP2, EGP40, TAG72, B7-H6, IL-13 receptor a2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, and an antigen associated with a universal tag, a cancer-testes antigen, mesothelin, MUC1, MUC16, PSCA, NKG2D Ligands, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), prostate specific antigen, PSMA, Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, and a pathogen-specific antigen. In some embodiments, the antigen is associated with or is a universal tag. In some embodiments, the antigen associated with the disease or disorder is selected from the group consisting of Notch 1, Notch 2, Notch 3, Notch 4, cell surface associated Mucin 1 (MUC1), Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CX3CR1, CXCL16, Delta1, E-cadherin, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R, and amyloid precursor protein (APP).

[0180] In some embodiments, the disease or disorder is a B cell-related disorder. In some embodiments, the disease or disorder is one or more diseases or conditions from among glioblastoma, lymphomatoid granulomatosis, post-transplant lymphoproliferative disorder, an immunoregulatory disorder, heavy-chain disease, primary or immunocyte-associated amyloidosis, or monoclonal gammopathy of undetermined significance.

[0181] In some embodiments, the disease or disorder is an autoimmune disease or disorder. Such autoimmune diseases or disorder include, but are not limited to, systemic lupus erythematosus (SLE), lupus nephritis, inflammatory bowel disease, rheumatoid arthritis (e.g., juvenile rheumatoid arthristis), ANCA associated vasculitis, idiopathic thrombocytopenia purpura (ITP), thrombotic thrombocytopenia purpura (TTP), autoimmune thrombocytopenia, Chagas' disease, Grave's disease, Wegener's granulomatosis, poly-arteritis nodosa, Sjogren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, psoriasis, IgA nephropathy, IgM polyneuropathies, vasculitis, diabetes mellitus, Reynaud's syndrome, anti-phospholipid syndrome, Goodpasture's disease, Kawasaki disease, autoimmune hemolytic anemia, myasthenia gravis, or progressive glomerulonephritis.

[0182] In some embodiments, the disease or disorder is a B cell malignancy. In some embodiments, the cancer (e.g., a BCMA-expressing cancer) is a lymphoma, a leukemia, or a plasma cell malignancy. Lymphomas contemplated herein include, but are not limited to, Burkitt lymphoma (e.g., endemic Burkitt's lymphoma or sporadic Burkitt's lymphoma), non-Hodgkin's lymphoma (NHL), Hodgkin's lymphoma, Waldenstrom macroglobulinemia, follicular lymphoma, small non-cleaved cell lymphoma, mucosa-associated lymphatic tissue lymphoma (MALT), marginal zone lymphoma, splenic lymphoma, nodal monocytoid B cell lymphoma, immunoblastic lymphoma, large cell lymphoma, diffuse mixed cell lymphoma, pulmonary B cell angiocentric lymphoma, small lymphocytic lymphoma, primary mediastinal B cell lymphoma, lymphoplasmacytic lymphoma (LPL), or mantle cell lymphoma (MCL). Leukemias contemplated herein, include, but are not limited to, chronic lymphocytic leukemia (CLL), plasma cell leukemia or acute lymphocytic leukemia (ALL). Also contemplated herein are plasma cell malignancies including, but not limited to, multiple myeloma (e.g., non-secretory multiple myeloma, smoldering multiple myeloma) or plasmacytoma. In some embodiments the disease or condition is multiple myeloma, such as relapsed and/or refractory multiple myeloma. Among the diseases, disorders or conditions that can be treated include, but are not limited to, neuroblastoma, renal cell carcinoma, colon cancer, colorectal cancer, breast cancer, epithelial squamous cell cancer, melanoma, myeloma (e.g., multiple myeloma), stomach cancer, brain cancer, lung cancer, pancreatic cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, prostate cancer, testicular cancer, thyroid cancer, uterine cancer, adrenal cancer and head and neck cancer.

[0183] In some embodiments, the cancer or proliferative disease is a B cell malignancy or hematological malignancy. In some embodiments the cancer or proliferative disease is acute lymphoblastic leukemia (ALL), non-Hodgkin's lymphoma (NHL), or chronic lymphocytic leukemia (CLL). In some embodiments, the cancer is CLL. In some embodiments, the methods can be used to treat a myeloma, a lymphoma or a leukemia. In some embodiments, the methods can be used to treat a non-Hodgkin lymphoma (NHL), an acute lymphoblastic leukemia (ALL), a chronic lymphocytic leukemia (CLL), a diffuse large B-cell lymphoma (DLBCL), acute myeloid leukemia (AML), or a myeloma, e.g., a multiple myeloma (MM). In some embodiments, the methods can be used to treat a MM or a DBCBL.

[0184] In some embodiments, the methods can be used to treat a non-hematologic cancer, such as a solid tumor. In some embodiments, the methods can be used to treat bladder, lung, brain, melanoma (e.g. small-cell lung, melanoma), breast, cervical, ovarian, colorectal, pancreatic, endometrial, esophageal, kidney, liver, prostate, skin, thyroid, or uterine cancers. In some embodiments, the cancer or proliferative disease is pancreatic cancer, bladder cancer, colorectal cancer, breast cancer, prostate cancer, renal cancer, hepatocellular cancer, lung cancer, ovarian cancer, cervical cancer, pancreatic cancer, rectal cancer, thyroid cancer, uterine cancer, gastric cancer, esophageal cancer, head and neck cancer, melanoma, neuroendocrine cancers, CNS cancers, brain tumors, bone cancer, or soft tissue sarcoma.

[0185] In some embodiments, the disease or condition is an infectious disease or condition, such as, but not limited to, viral, retroviral, bacterial, and protozoal infections, immunodeficiency, Cytomegalovirus (CMV), Epstein-Barr virus (EBV), adenovirus, BK polyomavirus. In some embodiments, the disease or condition is an autoimmune or inflammatory disease or condition, such as arthritis, e.g., rheumatoid arthritis (RA), Type I diabetes, systemic lupus erythematosus (SLE), inflammatory bowel disease, psoriasis, scleroderma, autoimmune thyroid disease, Graves disease, Crohn's disease, multiple sclerosis, asthma, and/or a disease or condition associated with transplant.

[0186] For the prevention or treatment of disease, the appropriate dosage of gamma secretase inhibitor and/or immunotherapy, such as a cell therapy (e.g. CAR-expressing T cells), may depend on the type of disease to be treated, cells and/or recombinant receptors expressed on the cells, the severity and course of the disease, route of administration, whether the gamma secretase inhibitor and/or the T cell therapy are administered for preventive or therapeutic purposes, previous therapy, frequency of administration, the subject's clinical history and response to the cells, and the discretion of the attending physician. The compositions and cells are in some embodiments suitably administered to the subject at one time or over a series of treatments. Exemplary dosage regimens and schedules for the provided combination therapy are described.

[0187] In some embodiments, the immunotherapy, such as cell therapy (e.g. CAR-T cell therapy) and the gamma secretase inhibitor are administered as part of a further combination treatment, which can be administered simultaneously with or sequentially to, in any order, another therapeutic intervention. In some contexts, the immunotherapy, such as cell therapy, e.g. engineered T cells, such as CAR-expressing T cells, are co-administered with another therapy sufficiently close in time such that the cell therapy enhances the effect of one or more additional therapeutic agents, or vice versa. In some embodiments, the cells are administered prior to the one or more additional therapeutic agents. In some embodiments, the cell therapy, e.g. engineered T cells, such as CAR-expressing T cells, are administered after the one or more additional therapeutic agents. In some embodiments, the combination therapy methods further include a lymphodepleting therapy, such as administration of a chemotherapeutic agent. In some embodiments, the combination therapy further comprises administering another therapeutic agent, such as an anti-cancer agent, a checkpoint inhibitor, or another immune modulating agent. In some embodiments, the combination therapy further comprises administering a steroid, such as dexamethasone. Uses include uses of the combination therapies in such methods and treatments, and uses of such compositions in the preparation of a medicament in order to carry out such combination therapy methods. In some embodiments, the methods and uses thereby treat the disease or condition or disorder, such as a cancer or proliferative disease, in the subject.

[0188] Prior to, during or following administration of the immunotherapy (e.g. T cell therapy, such as CAR-T cell therapy) and/or a gamma secretase inhibitor, the biological activity of the T cell therapy, e.g. the biological activity of the engineered cell populations, in some embodiments is measured, e.g., by any of a number of known methods. Parameters to assess include the ability of the engineered cells to destroy target cells, persistence and other measures of T cell activity, such as measured using any suitable method known in the art, such as assays described further below in Section III. In some embodiments, the biological activity of the cells, e.g., T cells administered for the T cell based therapy, is measured by assaying cytotoxic cell killing, expression and/or secretion of one or more cytokines, proliferation or expansion, such as upon restimulation with antigen. In some aspects the biological activity is measured by assessing the disease burden and/or clinical outcome, such as reduction in tumor burden or load. In some embodiments, administration of one or both agents of the combination therapy and/or any repeated administration of the therapy, can be determined based on the results of the assays before, during, during the course of or after administration of one or both agents of the combination therapy.

[0189] In some embodiments, the combined effect of the gamma secretase inhibitor in combination with the cell therapy can be synergistic compared to treatments involving only the gamma secretase inhibitor or monotherapy with the cell therapy. For example, in some embodiments, the methods provided herein result in an increase or an improvement in a desired therapeutic effect, such as an increased or an improvement in the reduction or inhibition of one or more symptoms associated with cancer.

[0190] In some embodiments, the gamma secretase inhibitor increases the expansion or proliferation of the engineered T cells, such as CAR T-Cells. In some embodiments, the increase in expansion or proliferation is observed in vivo upon administration to a subject. In some embodiments, the increase in the number of engineered T cells, e.g. CAR-T cells, is increased by greater than or greater than about 1.2-fold, 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 6.0-fold, 7.0-fold, 8.0-fold, 9.0-fold, 10.0 fold or more. In some embodiments, the gamma secretase inhibitor prolongs and/or sustains the activity and/or function of engineered T cells, such as CAR T-Cells.

[0191] A. Administration of an Immunotherapy

[0192] In some embodiments, an immunotherapy that binds to or targets an antigen on an immune cell and/or that is involved in a disease or disorder is administered in accord with the provided combination therapy methods. In particular embodiments, the immunotherapy binds to and/or recognizes an antigen that is expressed on or in a cell or tissue. In certain embodiments, the antigen is expressed on or in a cell or tissue. In particular embodiments, the antigen is expressed on the surface of a cell. In some embodiments, the cell is a B cell, a T cell and/or a tumor or cancer cell. In particular embodiments, the antigen is expressed in or on a circulating cell. In some embodiments, the antigen is expressed on the surface of a circulating cell.

[0193] In particular embodiments, the immunotherapy binds to and/or recognizes at least one antigen associated with a disease. Among the diseases, conditions, and disorders that may be treated in human subjects with the immunotherapy are tumors, including solid tumors, hematologic malignancies, and melanomas, and including localized and metastatic tumors, infectious diseases, such as infection with a virus or other pathogen, e.g., HIV, HCV, HBV, CMV, HPV, and parasitic disease, and autoimmune and inflammatory diseases. In some embodiments, the disease or condition is a tumor, cancer, malignancy, neoplasm, or other proliferative disease or disorder. Such diseases include but are not limited to leukemia, lymphoma, e.g., chronic lymphocytic leukemia (CLL), ALL, non-Hodgkin's lymphoma, acute myeloid leukemia, multiple myeloma, refractory follicular lymphoma, mantle cell lymphoma, indolent B cell lymphoma, B cell malignancies, cancers of the colon, lung, liver, breast, prostate, ovarian, skin, melanoma, bone, and brain cancer, ovarian cancer, epithelial cancers, renal cell carcinoma, pancreatic adenocarcinoma, Hodgkin lymphoma, cervical carcinoma, colorectal cancer, glioblastoma, neuroblastoma, Ewing sarcoma, medulloblastoma, osteosarcoma, synovial sarcoma, and/or mesothelioma.

[0194] In some embodiments, the immunotherapy binds to at least one antigen that is a target for cleavage by a gamma secretase. In some embodiments, the antigen is BCMA or is mucin 1 (Muc 1).

[0195] 1. Therapeutic Agents; e.g. Antibodies

[0196] In certain embodiments, the immunotherapy is a therapeutic agent, such as an antibody or antigen-binding fragment thereof. In some aspects, the antibody or antigen-binding fragment is a bispecific antibody.

[0197] In some aspects, the immunotherapy is a therapeutic agent that is or contains an immune system activator or stimulator. In certain embodiments, the immune system stimulator is an agent or therapy that activates at least one immune cell. In some embodiments, the immune cell is a T cell. In certain embodiments, the immune cell activator is IL-2, e.g., Proleukin; rhu-IFN-alpha-2a and/or rhu-IFN-alpha-2b, e.g., Pegasys, Roferon-A, Intron-A, and PEG intron; Anti-CD3 monoclonal antibody, e.g., Muromonab-CD3 and/or Orthoclone OKT 3; TGN-1412; and/or Blinatumomab, e.g., anti-CD3xCD3 BiTE.

[0198] In some embodiments, the immunotherapy is or contains a T cell-engaging therapy that is or comprises a binding molecule capable of binding to a surface molecule expressed on a T cell. In some embodiments, the surface molecule is an activating component of a T cell, such as a component of the T cell receptor complex. In some embodiments, the surface molecule is CD3 or is CD2. In some embodiments, the T cell-engaging therapy is or comprises an antibody or antigen-binding fragment. In some embodiments, the T cell-engaging therapy is a bispecific antibody containing at least one antigen-binding domain binding to an activating component of the T cell (e.g. a T cell surface molecule, e.g. CD3 or CD2) and at least one antigen-binding domain binding to a surface antigen on a target cell, such as a surface antigen on a tumor or cancer cell, for example any of the listed antigens as described herein, e.g. BCMA or Muc1. In some embodiments, the simultaneous or near simultaneous binding of such an antibody to both of its targets can result in a temporary interaction between the target cell and T cell, thereby resulting in activation, e.g. cytotoxic activity, of the T cell and subsequent lysis of the target cell.

[0199] Among such exemplary bispecific antibody T cell-engagers are bispecific T cell engager (BiTE) molecules, which contain tandem scFv molecules fused by a flexible linker (see e.g. Nagorsen and Bauerle, Exp Cell Res 317, 1255-1260 (2011); tandem scFv molecules fused to each other via, e.g. a flexible linker, and that further contain an Fc domain composed of a first and a second subunit capable of stable association (WO2013026837); diabodies and derivatives thereof, including tandem diabodies (Holliger et al, Prot Eng 9, 299-305 (1996); Kipriyanov et al, J Mol Biol 293, 41-66 (1999)); dual affinity retargeting (DART) molecules that can include the diabody format with a C-terminal disulfide bridge; or triomabs that include whole hybrid mouse/rat IgG molecules (Seimetz et al, Cancer Treat Rev 36, 458-467 (2010). In some embodiments, the T-cell engaging therapy is blinatumomab or AMG 330. Any of such T cell-engagers can be used in the provided methods, compositions or combinations.

[0200] In some embodiments, the therapeutic agent is a BCMA-specific binding capable of binding to BCMA and at least one additional antigen. In some embodiments, the at least one additional antigen CD19, CD20, CD22, CD33, CD38, CS1, ROR1, GPC3, CD123, IL-13R, CD138, c-Met, EGFRvIII, GD-2, NY-ESO-1, MAGE A3, or glycolipid F77. Examples of such bispecific antibodies are described in WO2017/025038.

[0201] The therapeutic agent, such as immunotherapy, can be administered by any suitable means, for example, by bolus infusion, by injection, e.g., intravenous or subcutaneous injections, intraocular injection, periocular injection, subretinal injection, intravitreal injection, trans-septal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjectval injection, subconjuntival injection, sub-Tenon's injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, the therapeutic agent, such as immunotherapy, is administered by parenteral, intrapulmonary, and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, intrathoracic, intracranial, or subcutaneous administration.

[0202] In certain embodiments, one or more doses of a T cell engaging therapy and/or an immune system stimulator are administered. In particular embodiments, between 0.001 .mu.g-5,000 .mu.g of the T cell engaging therapy and/or an immune system stimulator are administered. In particular embodiments, between 0.001 .mu.g to 1,000 .mu.g, 0.001 .mu.g to 1 .mu.g, 0.01 .mu.g to 1.mu.g, 0.1 .mu.g to10 .mu.g, 0.01 .mu.g to 1 .mu.g, 0.1 .mu.g to 5 .mu.g, 0.1 .mu.g to 50 .mu.g, 1 .mu.g to 100 .mu.g, 10 .mu.g to 100 .mu.g, 50 .mu.g to 500 .mu.g, 100 .mu.g to 1,000 .mu.g, 1,000 .mu.g to 2,000 .mu.g, or 2,000 .mu.g to 5,000 .mu.g of the T cell engaging therapy is administered. In some embodiments, the dose of the T cell engaging therapy is or includes between 0.01 .mu.g/kg and 100 .mu.g/kg, between 0.1 .mu.g/kg and 10 .mu.g/kg, between 10 .mu.g/kg and 50 .mu.g/kg, between 50 .mu.g/kg and 100 .mu.g/kg, between 0.1 mg/kg and 1 mg/kg, between 1 mg/kg and 10 mg/kg, between 10 mg/kg and 100 mg/kg, between 100 mg/kg and 500 mg/kg, between 200 mg/kg and 300 mg/kg, between 100 mg/kg and 250 mg/kg, between 200 mg/kg and 400 mg/kg, between 250 mg/kg and 500 mg/kg, between 250 mg/kg and 750 mg/kg, between 50 mg/kg and 750 mg/kg, between 1 mg/kg and 10 mg/kg, or between 100 mg/kg and 1,000 mg/kg (amount of the lymphodepleting agent over body weight). In some embodiments, the dose of the T cell engaging therapy is or is about 0.1 .mu.g/kg, 0.5 mg/kg, 1 .mu.g/kg, 5 .mu.g/kg, 10 .mu.g/kg, 20 .mu.g/kg, 30 .mu.g/kg, 40 .mu.g/kg, 50 .mu.g/kg, 60 .mu.g/kg, 70 .mu.g/kg, 80 .mu.g/kg, 90 .mu.g/kg, 0.1 mg/kg, 0.5 mg/kg, 1 mg/kg, 2.5 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 35 mg/kg, 40 mg/kg, 45 mg/kg, 50 mg/kg, 55 mg/kg, 60 mg/kg, 65 mg/kg, 70 mg/kg, 75 mg/kg, 80 mg/kg, 85 mg/kg, 90 mg/kg, 95 mg/kg, 100 mg/kg, 200 mg/kg, 300 mg/kg, 400 mg/kg, 500 mg/kg, 600 mg/kg, 700 mg/kg, 800 mg/kg, 900 mg/kg, or 1,000 mg/kg. In particular embodiments, the T cell engaging therapy is administered orally, intravenously, intraperitoneally, transdermally, intrathecally, intramuscularly, intranasally, transmucosally, subcutaneously, or rectally.

[0203] In some embodiments, the dose of CAR T cells administered can be reduced by 10%, 20%, 30%, 40%, 50%, 60%, 70%, or more of a dose of CAR T cells when administered in combination with a gamma secretase inhibitor. In some embodiments, the dose of CAR T cells administered when combined with gamma secretase inhibitor can be reduced by 10%, 20%, 30%, 40%, 50%, 60%, 70%, or more of a dose of CAR T cells administered without gamma secretase inhibitor.

[0204] 2. Cell Therapy

[0205] In some embodiments of the methods, compositions, combinations, kits and uses provided herein, the combination therapy includes administering to a subject an immune cell therapy, such as a T cell therapy (e.g. CAR-expressing T cells). Administration of such therapies can be initiated prior to, subsequent to, simultaneously with administration of the gamma secretase inhibitor as described. Exemplary cell therapies and engineered cells for use in the provided methods, compositions, combinations, kits and uses provided herein are described in Section II.

[0206] In some embodiments, the cell-based therapy is or comprises administration of cells, such as immune cells, for example T cell or NK cells, that target a molecule expressed on the surface of a lesion, such as a tumor or a cancer. In some embodiments, the immune cells express a T cell receptor (TCR) or other antigen-binding receptor. In some embodiments, the immune cells express a recombinant receptor, such as a transgenic TCR or a chimeric antigen receptor (CAR). In some embodiments, the cells are autologous to the subject. In some embodiments, the cells are allogeneic to the subject.

[0207] In some aspects, the cell therapy is or comprises a tumor infiltrating lymphocytic (TIL) therapy, a transgenic TCR therapy or a T cell therapy comprising genetically engineered cells, such as a recombinant-receptor expressing cell therapy. In some embodiments, the recombinant receptor specifically binds to a ligand, such as one associated with a disease or condition, e.g. associated with or expressed on a cell of a tumor or cancer. In some embodiments, the cell therapy includes administering T cells engineered to express a chimeric antigen receptor (CAR).

[0208] In some embodiments, the provided cells express and/or are engineered to express receptors, such as recombinant receptors, including those containing ligand-binding domains or binding fragments thereof, and T cell receptors (TCRs) and components thereof, and/or functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). In some embodiments, the recombinant receptor contains an extracellular ligand-binding domain that specifically binds to an antigen. In some embodiments, the recombinant receptor is a CAR that contains an extracellular antigen-recognition domain that specifically binds to an antigen. In some embodiments, the ligand, such as an antigen, is a protein expressed on the surface of cells. In some embodiments, the CAR is a TCR-like CAR and the antigen is a processed peptide antigen, such as a peptide antigen of an intracellular protein, which, like a TCR, is recognized on the cell surface in the context of a major histocompatibility complex (MHC) molecule.

[0209] Among the engineered cells, including engineered cells containing recombinant receptors, are described furtherherein. Exemplary recombinant receptors, including CARs and recombinant TCRs, as well as methods for engineering and introducing the receptors into cells, include those described, for example, in international patent application publication numbers WO200014257, WO2013126726, WO2012/129514, WO2014031687, WO2013/166321, WO2013/071154, WO2013/123061 U.S. patent application publication numbers US2002131960, US2013287748, US20130149337, U.S. Pat. Nos. 6,451,995, 7,446,190, 8,252,592, 8,339,645, 8,398,282, 7,446,179, 6,410,319, 7,070,995, 7,265,209, 7,354,762, 7,446,191, 8,324,353, and 8,479,118, and European patent application number EP2537416,and/or those described by Sadelain et al., Cancer Discov. 2013 April; 3(4): 388-398; Davila et al. (2013) PLoS ONE 8(4): e61338; Turtle et al., Curr. Opin. Immunol., 2012 October; 24(5): 633-39; Wu et al., Cancer, 2012 Mar. 18(2): 160-75. In some aspects, the genetically engineered antigen receptors include a CAR as described in U.S. Pat. No. 7,446,190, and those described in International Patent Application Publication No.: WO/2014055668 A1.

[0210] In some embodiments, the combination therapy includes administration to a subject cells, e.g. T cells, expressing a recombinant receptor that specifically recognize and/or target an antigen associated with the cancer and/or present on a universal tag. In some embodiments, the antigen recognized or targeted by the T cells is B cell maturation antigen (BCMA), ROR1,carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, optionally a human antigen of any of the foregoing; a pathogen-specific antigen. In some embodiments, the antigen recognized and/or targeted by T cells is selected from the group consisting of Notch 1, Notch 2, Notch 3, Notch 4, cell surface associated Mucin 1 (MUC1), Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CX3CR1, CXCL16, Delta1, E-cadherin, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R, and amyloid precursor protein (APP).

[0211] In some embodiments the antigen recognized and/targeted by T cells is B Cell Maturation Antigen (BCMA). Exemplary antigen-binding domains, and CARs containing such antigen-binding domains, that target or specifically bind BCMA are known, see e.g. WO 2016/090320, WO2016090327, WO2010104949A2 and WO2017173256. In some embodiments, the antigen binding domain is an scFv that contains a VH and a VL derived from an antibody or an antibody fragment specific to BCMA. In some embodiments, the antibody or antibody fragment that binds BCMA is or contains a VH and a VL from an antibody or antibody fragment set forth in International Patent Applications, Publication Number WO 2016/090327 and WO 2016/090320.

[0212] Methods for administration of engineered cells for adoptive cell therapy are known and may be used in connection with the provided methods and compositions. For example, adoptive T cell therapy methods are described, e.g., in US Patent Application Publication No. 2003/0170238 to Gruenberg et al; U.S. Pat. No. 4,690,915 to Rosenberg; Rosenberg (2011) Nat Rev Clin Oncol. 8(10):577-85). See, e.g., Themeli et al., (2013) Nat Biotechnol. 31(10): 928-933; Tsukahara et al., (2013) Biochem Biophys Res Commun 438(1): 84-9; Davila et al., (2013) PLoS ONE 8(4): e61338.

[0213] In some embodiments, the cell therapy, e.g., adoptive T cell therapy, is carried out by autologous transfer, in which the cells are isolated and/or otherwise prepared from the subject who is to receive the cell therapy, or from a sample derived from such a subject. Thus, in some aspects, the cells are derived from a subject, e.g., patient, in need of a treatment and the cells, following isolation and processing are administered to the same subject.

[0214] In some embodiments, the cell therapy, e.g., adoptive T cell therapy, is carried out by allogeneic transfer, in which the cells are isolated and/or otherwise prepared from a subject other than a subject who is to receive or who ultimately receives the cell therapy, e.g., a first subject. In such embodiments, the cells then are administered to a different subject, e.g., a second subject, of the same species. In some embodiments, the first and second subjects are genetically identical. In some embodiments, the first and second subjects are genetically similar. In some embodiments, the second subject expresses the same HLA class or supertype as the first subject.

[0215] The cells can be administered by any suitable means. The cells are administered in a dosing regimen to achieve a therapeutic effect, such as a reduction in tumor burden. Dosing and administration may depend in part on the schedule of administration of the gamma secretase inhibitor, which can be administered prior to, subsequent to and/or simultaneously with initiation of administration of the T cell therapy. Various dosing schedules of the T cell therapy include but are not limited to single or multiple administrations over various time-points, bolus administration, and pulse infusion.

[0216] a. Compositions and Formulations

[0217] In some embodiments, the dose of cells of the cell therapy, such a T cell therapy comprising cells engineered with a recombinant antigen receptor, e.g. CAR or TCR, is provided as a composition or formulation, such as a pharmaceutical composition or formulation. Such compositions can be used in accord with the provided methods, such as in the prevention or treatment of diseases, conditions, and disorders.

[0218] In some embodiments, the cell therapy, such as engineered T cells (e.g. CAR T cells), are formulated with a pharmaceutically acceptable carrier. In some aspects, the choice of carrier is determined in part by the particular cell or agent and/or by the method of administration. Accordingly, there are a variety of suitable formulations. For example, the pharmaceutical composition can contain preservatives. Suitable preservatives may include, for example, methylparaben, propylparaben, sodium benzoate, and benzalkonium chloride. In some aspects, a mixture of two or more preservatives is used. The preservative or mixtures thereof are typically present in an amount of about 0.0001% to about 2% by weight of the total composition. Carriers are described, e.g., by Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Pharmaceutically acceptable carriers are generally nontoxic to recipients at the dosages and concentrations employed, and include, but are not limited to: buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g. Zn-protein complexes); and/or non-ionic surfactants such as polyethylene glycol (PEG).

[0219] Buffering agents in some aspects are included in the compositions. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some aspects, a mixture of two or more buffering agents is used. The buffering agent or mixtures thereof are typically present in an amount of about 0.001% to about 4% by weight of the total composition. Methods for preparing administrable pharmaceutical compositions are known. Exemplary methods are described in more detail in, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins; 21st ed. (May 1, 2005).

[0220] The formulations can include aqueous solutions. The formulation or composition may also contain more than one active ingredient useful for the particular indication, disease, or condition being prevented or treated with the cells or agents, where the respective activities do not adversely affect one another. Such active ingredients are suitably present in combination in amounts that are effective for the purpose intended. Thus, in some embodiments, the pharmaceutical composition further includes other pharmaceutically active agents or drugs, such as chemotherapeutic agents, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.

[0221] The pharmaceutical composition in some embodiments contains cells in amounts effective to treat or prevent the disease or condition, such as a therapeutically effective or prophylactically effective amount. Therapeutic or prophylactic efficacy in some embodiments is monitored by periodic assessment of treated subjects. For repeated administrations over several days or longer, depending on the condition, the treatment is repeated until a desired suppression of disease symptoms occurs. However, other dosage regimens may be useful and can be determined. The desired dosage can be delivered by a single bolus administration of the composition, by multiple bolus administrations of the composition, or by continuous infusion administration of the composition.

[0222] The cells may be administered using standard administration techniques, formulations, and/or devices. Provided are formulations and devices, such as syringes and vials, for storage and administration of the compositions. With respect to cells, administration can be autologous or heterologous. For example, immunoresponsive cells or progenitors can be obtained from one subject, and administered to the same subject or a different, compatible subject. Peripheral blood derived immunoresponsive cells or their progeny (e.g., in vivo, ex vivo or in vitro derived) can be administered via localized injection, including catheter administration, systemic injection, localized injection, intravenous injection, or parenteral administration. When administering a therapeutic composition (e.g., a pharmaceutical composition containing a genetically modified immunoresponsive cell), it will generally be formulated in a unit dosage injectable form (solution, suspension, emulsion).

[0223] Formulations include those for oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. In some embodiments, the agent or cell populations are administered parenterally. The term "parenteral," as used herein, includes intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal administration. In some embodiments, the agent or cell populations are administered to a subject using peripheral systemic delivery by intravenous, intraperitoneal, or subcutaneous injection.

[0224] Compositions in some embodiments are provided as sterile liquid preparations, e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may in some aspects be buffered to a selected pH. Liquid preparations are normally easier to prepare than gels, other viscous compositions, and solid compositions. Additionally, liquid compositions are somewhat more convenient to administer, especially by injection. Viscous compositions, on the other hand, can be formulated within the appropriate viscosity range to provide longer contact periods with specific tissues. Liquid or viscous compositions can comprise carriers, which can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, polyoi (for example, glycerol, propylene glycol, liquid polyethylene glycol) and suitable mixtures thereof.

[0225] Sterile injectable solutions can be prepared by incorporating the cells in a solvent, such as in admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose, dextrose, or the like. The compositions can also be lyophilized. The compositions can contain auxiliary substances such as wetting, dispersing, or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or viscosity enhancing additives, preservatives, flavoring agents, colors, and the like, depending upon the route of administration and the preparation desired. Standard texts may in some aspects be consulted to prepare suitable preparations.

[0226] Various additives which enhance the stability and sterility of the compositions, including antimicrobial preservatives, antioxidants, chelating agents, and buffers, can be added. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0227] The formulations to be used for in vivo administration are generally sterile. Sterility may be readily accomplished, e.g., by filtration through sterile filtration membranes.

[0228] For the prevention or treatment of disease, the appropriate dosage may depend on the type of disease to be treated, the type of agent or agents, the type of cells or recombinant receptors, the severity and course of the disease, whether the agent or cells are administered for preventive or therapeutic purposes, previous therapy, the subject's clinical history and response to the agent or the cells, and the discretion of the attending physician. The compositions are in some embodiments suitably administered to the subject at one time or over a series of treatments.

[0229] In some cases, the cell therapy is administered as a single pharmaceutical composition comprising the cells. In some embodiments, a given dose is administered by a single bolus administration of the cells or agent. In some embodiments, it is administered by multiple bolus administrations of the cells or agent, for example, over a period of no more than 3 days, or by continuous infusion administration of the cells or agent.

[0230] b. Dosage Schedule and Administration

[0231] In some embodiments, a dose of cells is administered to subjects in accord with the provided combination therapy methods. In some embodiments, the size or timing of the doses is determined as a function of the particular disease or condition in the subject. It is within the level of a skilled artisan to empirically determine the size or timing of the doses for a particular disease in view of the provided description.

[0232] In certain embodiments, the cells, or individual populations of sub-types of cells, are administered to the subject at a range of about 0.1 million to about 100 billion cells and/or that amount of cells per kilogram of body weight of the subject, such as, e.g., 0.1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), 1 million to about 50 billion cells (e.g., about 5 million cells, about 25 million cells, about 500 million cells, about 1 billion cells, about 5 billion cells, about 20 billion cells, about 30 billion cells, about 40 billion cells, or a range defined by any two of the foregoing values), such as about 10 million to about 100 billion cells (e.g., about 20 million cells, about 30 million cells, about 40 million cells, about 60 million cells, about 70 million cells, about 80 million cells, about 90 million cells, about 10 billion cells, about 25 billion cells, about 50 billion cells, about 75 billion cells, about 90 billion cells, or a range defined by any two of the foregoing values), and in some cases about 100 million cells to about 50 billion cells (e.g., about 120 million cells, about 250 million cells, about 350 million cells, about 450 million cells, about 650 million cells, about 800 million cells, about 900 million cells, about 3 billion cells, about 30 billion cells, about 45 billion cells) or any value in between these ranges and/or per kilogram of body weight of the subject. Dosages may vary depending on attributes particular to the disease or disorder and/or patient and/or other treatments. In some embodiments, such values refer to numbers of recombinant receptor-expressing cells; in other embodiments, they refer to number of T cells or PBMCs or total cells administered.

[0233] In some embodiments, for example, where the subject is a human, the cells are administered at a dose of total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs) that is in the range of about 1.times.10.sup.6 to about 1.2.times.10.sup.9 such cells, such as at or about 1.times.10.sup.7, at or about 5.times.10.sup.7, at or about 1.times.10.sup.8, at or about 2.times.10.sup.8, at or about 2.5.times.10.sup.8, at or about 3.times.10.sup.8, at or about 3.5.times.10.sup.8, at or about 4.times.10.sup.8, at or about 4.5.times.10.sup.8 at or about 5.times.10.sup.8 total such cells, at or about 6.times.10.sup.8, at or about 7.times.10.sup.8, at or about 8.times.10.sup.8, at or about 9.times.10.sup.8 or at or about 1.times.10.sup.9 of such cells, or the range between any two of the foregoing values. In some embodiments, for example, where the subject is a human, the dose includes fewer than about or about 5.times.10.sup.8 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes fewer than about or about 3.0.times.10.sup.8 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes fewer than about or about 1.5.times.10.sup.8 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes fewer than about 1.times.10.sup.8 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs). In some embodiments, for example, where the subject is a human, the dose includes fewer than about 5.times.10.sup.7 total recombinant receptor (e.g., CAR)-expressing cells, T cells, or peripheral blood mononuclear cells (PBMCs).

[0234] In some embodiments, the dose of total cells administered comprises from or from about 1.times.10.sup.5 to 8.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.5 to 5.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.5 to 2.5.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.5 to 1.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.5 to 5.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.5 to 2.5.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.5 to 1.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.5 to 5.times.10.sup.6 total CAR-expressing T cells, 1.times.10.sup.5 to 2.5.times.10.sup.6 total CAR-expressing T cells, 1.times.10.sup.5 to 1.times.10.sup.6 total CAR-expressing T cells, 1.times.10.sup.6 to 8.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.6 to 5.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.6 to 2.5.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.6 to 1.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.6 to 5.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.6 to 2.5.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.6 to 1.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.6 to 5.times.10.sup.6 total CAR-expressing T cells, 1.times.10.sup.6 to 2.5.times.10.sup.6 total CAR-expressing T cells, 2.5.times.10.sup.6 to 8.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.6 to 5.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.6 to 2.5.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.6 to 1.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.6 to 5.times.10.sup.7 total CAR-expressing T cells, 2.5.times.10.sup.6 to 2.5.times.10.sup.7 total CAR-expressing T cells, 2.5.times.10.sup.6 to 1.times.10.sup.7 total CAR-expressing T cells, 2.5.times.10.sup.6 to 5.times.10.sup.6 total CAR-expressing T cells, 5.times.10.sup.6 to 8.times.10.sup.8 total CAR-expressing T cells, 5.times.10.sup.6 to 5.times.10.sup.8 total CAR-expressing T cells, 5.times.10.sup.6 to 2.5.times.10.sup.8 total CAR-expressing T cells, 5.times.10.sup.6 to 1.times.10.sup.8 total CAR-expressing T cells, 5.times.10.sup.6 to 5.times.10.sup.7 total CAR-expressing T cells, 5.times.10.sup.6 to 2.5.times.10.sup.7 total CAR-expressing T cells, 5.times.10.sup.6 to 1.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.7 to 8.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.7 to 5.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.7 to 2.5.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.7 to 1.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.7 to 5.times.10.sup.7 total CAR-expressing T cells, 1.times.10.sup.7 to 2.5.times.10.sup.7 total CAR-expressing T cells, 2.5.times.10.sup.7 to 8.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.7 to 5.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.7 to 2.5.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.7 to 1.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.7 to 5.times.10.sup.7 total CAR-expressing T cells, 5.times.10.sup.7 to 8.times.10.sup.8 total CAR-expressing T cells, 5.times.10.sup.7 to 5.times.10.sup.8 total CAR-expressing T cells, 5.times.10.sup.7 to 2.5.times.10.sup.8 total CAR-expressing T cells, 5.times.10.sup.7 to 1.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.8 to 8.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.8 to 5.times.10.sup.8 total CAR-expressing T cells, 1.times.10.sup.8 to 2.5.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.8 to 8.times.10.sup.8 total CAR-expressing T cells, 2.5.times.10.sup.8 to 5.times.10.sup.8 total CAR-expressing T cells, or 5.times.10.sup.8 total CAR-expressing T cells to 8.times.10.sup.8 total CAR-expressing T cells, each inclusive.

[0235] In some embodiments, the dose of genetically engineered cells comprises from or from about 50.times.10.sup.6 to 800.times.10.sup.6 total CAR-expressing T cells, from or from about 50.times.10.sup.6 to 450.times.10.sup.6 total CAR-expressing T cells, from or from about 50.times.10.sup.6 to 300.times.10.sup.6 total CAR-expressing T cells, from or from about 50.times.10.sup.6 to 150.times.10.sup.6 total CAR-expressing T cells, from or from about 150.times.10.sup.6 to 800.times.10.sup.6 total CAR-expressing T cells, from or from about 150.times.10.sup.6 to 450.times.10.sup.6 total CAR-expressing T cells, from or from about 150.times.10.sup.6 to 300.times.10.sup.6 total CAR-expressing T cells, 300.times.10.sup.6 to 800.times.10.sup.6 total CAR-expressing T cells, from or from about 300.times.10.sup.6 to 450.times.10.sup.6 total CAR-expressing T cells, or 450.times.10.sup.6 to 800.times.10.sup.6 total CAR-expressing T cells. In some embodiments, the dose of genetically engineered cells is or is about 50.times.10.sup.6 CAR-expressing T cells, is or is about 150.times.10.sup.6 CAR-expressing T cells, cells, is or is about 450.times.10.sup.6 CAR-expressing T cells, or is or is about 800.times.10.sup.6 CAR-expressing T cells.

[0236] In some of any of the embodiments, the number is with reference to the total number of CD3+, total number of CD8+ or total number of CD4+ and CD8+ T cells, in some recombinant receptor-expressing (e.g. CAR+) cells of such cells.

[0237] In some embodiments, the cell therapy comprises administration of a dose comprising a number of cells from or from about 1.times.10.sup.5 to 1.times.10.sup.8 total recombinant receptor-expressing cells, total T cells, or total peripheral blood mononuclear cells (PBMCs); from or from about 5.times.10.sup.5 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs or from or from about 1.times.10.sup.6 to 1.times.10.sup.7 total recombinant receptor-expressing cells, total T cells, or total PBMCs), each inclusive. In some embodiments, the cell therapy comprises administration of a dose of cells comprising a number of cells at least or about at least 1.times.10.sup.5 total recombinant receptor-expressing cells, total T cells, or total PBMCs, such at least or at least 1.times.10.sup.6, at least or about at least 1.times.10.sup.7, at least or about at least 1.times.10.sup.8 of such cells. In some embodiments, the number is with reference to the total number of CD3+ or CD8+, in some cases also recombinant receptor-expressing (e.g. CAR+) cells. In some embodiments, the cell therapy comprises administration of a dose comprising a number of cells from or from about 1.times.10.sup.5 to 1.times.10.sup.8 CD3+ or CD8+ total T cells or CD3+ or CD8+ recombinant receptor-expressing cells, from or from about 5.times.10.sup.5 to 1.times.10.sup.7 CD3+ or CD8+ total T cells or CD3+ or CD8+ recombinant receptor-expressing cells, or from or from about 1.times.10.sup.6 to 1.times.10.sup.7 CD3+ or CD8+ total T cells or CD3+ or CD8+recombinant receptor-expressing cells, each inclusive. In some embodiments, the cell therapy comprises administration of a dose comprising a number of cells from or from about 1.times.10.sup.5 to 1.times.10.sup.8 total CD3+/CAR+ or CD8+/CAR+ cells, from or from about 5.times.10.sup.5 to 1.times.10.sup.7 total CD3+/CAR+ or CD8+/CAR+ cells, or from or from about 1.times.10.sup.6 to 1.times.10.sup.7 total CD3+/CAR+ or CD8+/CAR+ cells, each inclusive.

[0238] In some embodiments, the T cells of the dose include CD4+ T cells, CD8+ T cells or CD4+ and CD8+ T cells.

[0239] In some embodiments, for example, where the subject is human, the CD8+ T cells of the dose, including in a dose including CD4+ and CD8+ T cells, includes between about 1.times.10.sup.6 and 5.times.10.sup.8 total recombinant receptor (e.g., CAR)-expressing CD8+cells, e.g., in the range of about 5.times.10.sup.6 to 1.times.10.sup.8 such cells, such cells 1.times.10.sup.7, 2.5.times.10.sup.7, 5.times.10.sup.7, 7.5.times.10.sup.7, 1.times.10.sup.8, or 5.times.10.sup.8 total such cells, or the range between any two of the foregoing values. In some embodiments, the patient is administered multiple doses, and each of the doses or the total dose can be within any of the foregoing values. In some embodiments, the dose of cells comprises the administration of from or from about 1.times.10.sup.7 to 0.75.times.10.sup.8 total recombinant receptor-expressing CD8+ T cells, 1.times.10.sup.7 to 2.5.times.10.sup.7 total recombinant receptor-expressing CD8+ T cells, from or from about 1.times.10.sup.7 to 0.75.times.10.sup.8 total recombinant receptor-expressing CD8+ T cells, each inclusive. In some embodiments, the dose of cells comprises the administration of or about 1.times.10.sup.7, 2.5.times.10.sup.7, 5.times.10.sup.7 7.5.times.10.sup.7, 1.times.10.sup.8, or 5.times.10.sup.8 total recombinant receptor-expressing CD8+ T cells.

[0240] In some embodiments, the cell therapy comprises administration of a dose comprising a number of cells that is at least or at least about or is or is about 0.1.times.10.sup.6 cells/kg body weight of the subject, 0.2.times.10.sup.6 cells/kg, 0.3.times.10.sup.6 cells/kg, 0.4.times.10.sup.6 cells/kg, 0.5.times.10.sup.6 cells/kg, 1.times.10.sup.6 cell/kg, 2.0.times.10.sup.6 cells/kg, 3.times.10.sup.6 cells/kg or 5.times.10.sup.6 cells/kg.

[0241] In some embodiments, the cell therapy comprises administration of a dose comprising a number of cells between or between about 0.1.times.10.sup.6 cells/kg body weight of the subject and 1.0.times.10.sup.7 cells/kg, between or between about 0.5.times.10.sup.6 cells/kg and 5.times.10.sup.6 cells/kg, between or between about 0.5.times.10.sup.6 cells/kg and 3.times.10.sup.6 cells/kg, between or between about 0.5.times.10.sup.6 cells/kg and 2.times.10.sup.6 cells/kg, between or between about 0.5.times.10.sup.6 cells/kg and 1.times.10.sup.6 cell/kg, between or between about 1.0.times.10.sup.6 cells/kg body weight of the subject and 5.times.10.sup.6 cells/kg, between or between about 1.0.times.10.sup.6 cells/kg and 3.times.10.sup.6 cells/kg, between or between about 1.0.times.10.sup.6 cells/kg and 2.times.10.sup.6 cells/kg, between or between about 2.0.times.10.sup.6 cells/kg body weight of the subject and 5.times.10.sup.6 cells/kg, between or between about 2.0.times.10.sup.6 cells/kg and 3.times.10.sup.6 cells/kg, or between or between about 3.0.times.10.sup.6 cells/kg body weight of the subject and 5.times.10.sup.6 cells/kg, each inclusive.

[0242] In some embodiments, the dose of cells comprises between at or about 2.times.10.sup.5 of the cells/kg and at or about 2.times.10.sup.6 of the cells/kg, such as between at or about 4.times.10.sup.5 of the cells/kg and at or about 1.times.10.sup.6 of the cells/kg or between at or about 6.times.10.sup.5 of the cells/kg and at or about 8.times.10.sup.5 of the cells/kg. In some embodiments, the dose of cells comprises no more than 2.times.10.sup.5 of the cells (e.g. antigen-expressing, such as CAR-expressing cells) per kilogram body weight of the subject (cells/kg), such as no more than at or about 3.times.10.sup.5 cells/kg, no more than at or about 4.times.10.sup.5 cells/kg, no more than at or about 5.times.10.sup.5 cells/kg, no more than at or about 6.times.10.sup.5 cells/kg, no more than at or about 7.times.10.sup.5 cells/kg, no more than at or about 8.times.10.sup.5 cells/kg, nor more than at or about 9.times.10.sup.5 cells/kg, no more than at or about 1.times.10.sup.6 cells/kg, or no more than at or about 2.times.10.sup.6 cells/kg. In some embodiments, the dose of cells comprises at least or at least about or at or about 2.times.10.sup.5 of the cells (e.g. antigen-expressing, such as CAR-expressing cells) per kilogram body weight of the subject (cells/kg), such as at least or at least about or at or about 3.times.10.sup.5 cells/kg, at least or at least about or at or about 4.times.10.sup.5 cells/kg, at least or at least about or at or about 5.times.10.sup.5 cells/kg, at least or at least about or at or about 6.times.10.sup.5 cells/kg, at least or at least about or at or about 7.times.10.sup.5 cells/kg, at least or at least about or at or about 8.times.10.sup.5 cells/kg, at least or at least about or at or about 9.times.10.sup.5 cells/kg, at least or at least about or at or about 1.times.10.sup.6 cells/kg, or at least or at least about or at or about 2.times.10.sup.6 cells/kg.

[0243] In some embodiments, the dose of cells, e.g., recombinant receptor-expressing T cells, is administered to the subject as a single dose or is administered only one time within a period of two weeks, one month, three months, six months, 1 year or more.

[0244] In some aspects, the pharmaceutical compositions and formulations are provided as unit dose form compositions including the number of cells for administration in a given dose or fraction thereof.

[0245] In some embodiments, cells of the dose may be administered by administration of a plurality of compositions or solutions, such as a first and a second, optionally more, each containing some cells of the dose. In some aspects, the plurality of compositions, each containing a different population and/or sub-types of cells, are administered separately or independently, optionally within a certain period of time. For example, the populations or sub-types of cells can include CD8.sup.+ and CD4.sup.+ T cells, respectively, and/or CD8+- and CD4+-enriched populations, respectively, e.g., CD4+ and/or CD8+ T cells each individually including cells genetically engineered to express the recombinant receptor. In some embodiments, the administration of the dose comprises administration of a first composition comprising a dose of CD8+ T cells or a dose of CD4+ T cells and administration of a second composition comprising the other of the dose of CD4+ T cells and the CD8+ T cells.

[0246] In some embodiments, the administration of the composition or dose, e.g., administration of the plurality of cell compositions, involves administration of the cell compositions separately. In some aspects, the separate administrations are carried out simultaneously, or sequentially, in any order. In some embodiments, the dose comprises a first composition and a second composition, and the first composition and second composition are administered 0 to 12 hours apart, 0 to 6 hours apart or 0 to 2 hours apart. In some embodiments, the initiation of administration of the first composition and the initiation of administration of the second composition are carried out no more than 2 hours, no more than 1 hour, or no more than 30 minutes apart, no more than 15 minutes, no more than 10 minutes or no more than 5 minutes apart. In some embodiments, the initiation and/or completion of administration of the first composition and the completion and/or initiation of administration of the second composition are carried out no more than 2 hours, no more than 1 hour, or no more than 30 minutes apart, no more than 15 minutes, no more than 10 minutes or no more than 5 minutes apart.

[0247] In some composition, the first composition, e.g., first composition of the dose, comprises CD4+ T cells. In some composition, the first composition, e.g., first composition of the dose, comprises CD8+ T cells. In some embodiments, the first composition is administered prior to the second composition.

[0248] In some embodiments, the dose or composition of cells includes a defined or target ratio of CD4+ cells expressing a recombinant receptor to CD8+ cells expressing a recombinant receptor and/or of CD4+ cells to CD8+ cells, which ratio optionally is approximately 1:1 or is between approximately 1:3 and approximately 3:1, such as approximately 1:1. In some aspects, the administration of a composition or dose with the target or desired ratio of different cell populations (such as CD4+:CD8+ ratio or CAR+CD4+:CAR+CD8+ ratio, e.g., 1:1) involves the administration of a cell composition containing one of the populations and then administration of a separate cell composition comprising the other of the populations, where the administration is at or approximately at the target or desired ratio. In some aspects, administration of a dose or composition of cells at a defined ratio leads to improved expansion, persistence and/or antitumor activity of the T cell therapy.

[0249] In the context of adoptive cell therapy, administration of a given "dose" of cells encompasses administration of the given amount or number of cells as a single composition and/or single uninterrupted administration, e.g., as a single injection or continuous infusion, and also encompasses administration of the given amount or number of cells as a split dose, provided in multiple individual compositions or infusions, over a specified period of time, which is no more than 3 days. Thus, in some contexts, the dose is a single or continuous administration of the specified number of cells, given or initiated at a single point in time. In some contexts, however, the dose is administered in multiple injections or infusions over a period of no more than three days, such as once a day for three days or for two days or by multiple infusions over a single day period.

[0250] Thus, in some aspects, the cells of the dose are administered in a single pharmaceutical composition. In some embodiments, the cells of the dose are administered in a plurality of compositions, collectively containing the cells of the dose.

[0251] The term "split dose" refers to a dose that is split so that it is administered over more than one day. This type of dosing is encompassed by the present methods and is considered to be a single dose. In some embodiments, the cells of a split dose are administered in a plurality of compositions, collectively comprising the cells of the dose, over a period of no more than three days.

[0252] Thus, the dose of cells may be administered as a split dose. For example, in some embodiments, the dose may be administered to the subject over 2 days or over 3 days. Exemplary methods for split dosing include administering 25% of the dose on the first day and administering the remaining 75% of the dose on the second day. In other embodiments, 33% of the dose may be administered on the first day and the remaining 67% administered on the second day. In some aspects, 10% of the dose is administered on the first day, 30% of the dose is administered on the second day, and 60% of the dose is administered on the third day. In some embodiments, the split dose is not spread over more than 3 days.

[0253] In some embodiments, the subject receives multiple doses, e.g., two or more doses or multiple consecutive doses, of the cells. In some embodiments, two doses are administered to a subject. In some embodiments, the subject receives the consecutive dose, e.g., second dose, is administered approximately 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21 days after the first dose. In some embodiments, multiple consecutive doses are administered following the first dose, such that an additional dose or doses are administered following administration of the consecutive dose. In some aspects, the number of cells administered to the subject in the additional dose is the same as or similar to the first dose and/or consecutive dose. In some embodiments, the additional dose or doses are larger than prior doses.

[0254] In some aspects, the size of the first and/or consecutive dose is determined based on one or more criteria such as response of the subject to prior treatment, e.g. chemotherapy, disease burden in the subject, such as tumor load, bulk, size, or degree, extent, or type of metastasis, stage, and/or likelihood or incidence of the subject developing toxic outcomes, e.g., CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and/or a host immune response against the cells and/or recombinant receptors being administered.

[0255] In some aspects, the time between the administration of the first dose and the administration of the consecutive dose is about 9 to about 35 days, about 14 to about 28 days, or 15 to 27 days. In some embodiments, the administration of the consecutive dose is at a time point more than about 14 days after and less than about 28 days after the administration of the first dose. In some aspects, the time between the first and consecutive dose is about 21 days. In some embodiments, an additional dose or doses, e.g. consecutive doses, are administered following administration of the consecutive dose. In some aspects, the additional consecutive dose or doses are administered at least about 14 and less than about 28 days following administration of a prior dose. In some embodiments, the additional dose is administered less than about 14 days following the prior dose, for example, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 days after the prior dose. In some embodiments, no dose is administered less than about 14 days following the prior dose and/or no dose is administered more than about 28 days after the prior dose.

[0256] In some embodiments, the dose of cells, e.g., recombinant receptor-expressing cells, comprises two doses (e.g., a double dose), comprising a first dose of the T cells and a consecutive dose of the T cells, wherein one or both of the first dose and the second dose comprises administration of the split dose of T cells.

[0257] In some embodiments, the dose of cells is generally large enough to be effective in reducing disease burden.

[0258] In some embodiments, the cells are administered at a desired dosage, which in some aspects includes a desired dose or number of cells or cell type(s) and/or a desired ratio of cell types. Thus, the dosage of cells in some embodiments is based on a total number of cells (or number per kg body weight) and a desired ratio of the individual populations or sub-types, such as the CD4+ to CD8+ ratio. In some embodiments, the dosage of cells is based on a desired total number (or number per kg of body weight) of cells in the individual populations or of individual cell types. In some embodiments, the dosage is based on a combination of such features, such as a desired number of total cells, desired ratio, and desired total number of cells in the individual populations.

[0259] In some embodiments, the populations or sub-types of cells, such as CD8.sup.+ and CD4.sup.+ T cells, are administered at or within a tolerated difference of a desired dose of total cells, such as a desired dose of T cells. In some aspects, the desired dose is a desired number of cells or a desired number of cells per unit of body weight of the subject to whom the cells are administered, e.g., cells/kg. In some aspects, the desired dose is at or above a minimum number of cells or minimum number of cells per unit of body weight. In some aspects, among the total cells administered at the desired dose, the individual populations or sub-types are present at or near a desired output ratio (such as CD4.sup.+ to CD8.sup.+ ratio), e.g., within a certain tolerated difference or error of such a ratio.

[0260] In some embodiments, the cells are administered at or within a tolerated difference of a desired dose of one or more of the individual populations or sub-types of cells, such as a desired dose of CD4+ cells and/or a desired dose of CD8+ cells. In some aspects, the desired dose is a desired number of cells of the sub-type or population, or a desired number of such cells per unit of body weight of the subject to whom the cells are administered, e.g., cells/kg. In some aspects, the desired dose is at or above a minimum number of cells of the population or sub-type, or minimum number of cells of the population or sub-type per unit of body weight.

[0261] Thus, in some embodiments, the dosage is based on a desired fixed dose of total cells and a desired ratio, and/or based on a desired fixed dose of one or more, e.g., each, of the individual sub-types or sub-populations. Thus, in some embodiments, the dosage is based on a desired fixed or minimum dose of T cells and a desired ratio of CD4.sup.+ to CD8.sup.+ cells, and/or is based on a desired fixed or minimum dose of CD4.sup.+ and/or CD8.sup.+ cells.

[0262] In some embodiments, the cells are administered at or within a tolerated range of a desired output ratio of multiple cell populations or sub-types, such as CD4+ and CD8+ cells or sub-types. In some aspects, the desired ratio can be a specific ratio or can be a range of ratios. For example, in some embodiments, the desired ratio (e.g., ratio of CD4.sup.+ to CD8.sup.+ cells) is between at or about 5:1 and at or about 5:1 (or greater than about 1:5 and less than about 5:1), or between at or about 1:3 and at or about 3:1 (or greater than about 1:3 and less than about 3:1), such as between at or about 2:1 and at or about 1:5 (or greater than about 1:5 and less than about 2:1, such as at or about 5:1, 4.5:1, 4:1, 3.5:1, 3:1, 2.5:1, 2:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9: 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, or 1:5. In some aspects, the tolerated difference is within about 1%, about 2%, about 3%, about 4% about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50% of the desired ratio, including any value in between these ranges.

[0263] The cells can be administered by any suitable means. The cells are administered in a dosing regimen to achieve a therapeutic effect, such as a reduction in tumor burden. Dosing and administration may depend in part on the schedule of administration of the immunomodulatory compound, which can be administered prior to, subsequent to and/or simultaneously with initiation of administration of the T cell therapy. Various dosing schedules of the T cell therapy include but are not limited to single or multiple administrations over various time-points, bolus administration, and pulse infusion. In certain embodiments, the engineered T cells express a recombinant receptor. In certain embodiments, the engineered T cells express a CAR.

[0264] In particular embodiments, the numbers and/or concentrations of cells refer to the number of recombinant receptor (e.g., CAR)-expressing cells. In other embodiments, the numbers and/or concentrations of cells refer to the number or concentration of all cells, T cells, or peripheral blood mononuclear cells (PBMCs) administered.

[0265] In some aspects, the size of the dose is determined based on one or more criteria such as response of the subject to prior treatment, e.g. chemotherapy, disease burden in the subject, such as tumor load, bulk, size, or degree, extent, or type of metastasis, stage, and/or likelihood or incidence of the subject developing toxic outcomes, e.g., CRS, macrophage activation syndrome, tumor lysis syndrome, neurotoxicity, and/or a host immune response against the cells and/or recombinant receptors being administered.

[0266] In some embodiments, administration of the gamma secretase inhibitor in combination with the cells is able to increase, such as substantially or significantly increase, the expansion or proliferation of the cells, and thus a lower dose of cells can be administered to the subject. In some cases, the provided methods allow a lower dose of such cells to be administered, to achieve the same or better efficacy of treatment as the dose in a method in which the cell therapy is administered without administering the gamma secretase inhibitor, such as at least 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold or 10-fold less than the dose in a method in which the cell therapy is administered without administering the gamma secretase inhibitor.

[0267] In some embodiments, for example, the dose contains between or between about 5.0.times.10.sup.6 and 2.25.times.10.sup.7, 5.0.times.10.sup.6 and 2.0.times.10.sup.7, 5.0.times.10.sup.6 and 1.5.times.10.sup.7, 5.0.times.10.sup.6 and 1.0.times.10.sup.7, 5.0.times.10.sup.6 and 7.5.times.10.sup.6, 7.5.times.10.sup.6 and 2.25.times.10.sup.7, 7.5.times.10.sup.6 and 2.0.times.10.sup.7, 7.5.times.10.sup.6 and 1.5.times.10.sup.7, 7.5.times.10.sup.6 and 1.0.times.10.sup.7, 1.0.times.10.sup.7 and 2.25.times.10.sup.7, 1.0.times.10.sup.7 and 2.0.times.10.sup.7, 1.0.times.10.sup.7 and 1.5.times.10.sup.7, 1.5.times.10.sup.7 and 2.25.times.10.sup.7, 1.5.times.10.sup.7 and 2.0.times.10.sup.7, 2.0.times.10.sup.7 and 2.25.times.10.sup.7. In some embodiments, the dose of cells contains a number of cells, that is between at least or at least about 5.times.10.sup.6, 6.times.10.sup.6, 7.times.10.sup.6, 8.times.10.sup.6, 9.times.10.sup.6, 10.times.10.sup.6 and about 15.times.10.sup.6 recombinant-receptor expressing cells, such as recombinant-receptor expressing cells that are CD8+. In some embodiments, such dose, such as such target number of cells refers to the total recombinant-receptor expressing cells in the administered composition.

[0268] In some embodiments, for example, the lower dose contains less than about 5.times.10.sup.6 cells, recombinant receptor (e.g. CAR)-expressing cells, T cells, and/or PBMCs per kilogram body weight of the subject, such as less than about 4.5.times.10.sup.6, 4.times.10.sup.6, 3.5.times.10.sup.6, 3.times.10.sup.6, 2.5.times.10.sup.6, 2.times.10.sup.6, 1.5.times.10.sup.6, 1.times.10.sup.6, 5.times.10.sup.5, 2.5.times.10.sup.5, or 1.times.10.sup.5 such cells per kilogram body weight of the subject. In some embodiments, the lower dose contains less than about 1.times.10.sup.5, 2.times.10.sup.5, 5.times.10.sup.5, or 1.times.10.sup.6 of such cells per kilogram body weight of the subject, or a value within the range between any two of the foregoing values. In some embodiments, such values refer to numbers of recombinant receptor-expressing cells; in other embodiments, they refer to the number of T cells or PBMCs or total cells administered.

[0269] In some embodiments, one or more subsequent dose of cells can be administered to the subject. In some embodiments, the subsequent dose of cells is administered greater than or greater than about 7 days, 14 days, 21 days, 28 days or 35 days after initiation of administration of the first dose of cells. The subsequent dose of cells can be more than, approximately the same as, or less than the first dose. In some embodiments, administration of the T cell therapy, such as administration of the first and/or second dose of cells, can be repeated.

[0270] In some embodiments, initiation of administration of the cell therapy, e.g. the dose of cells or a first dose of a split dose of cells, is administered before (prior to), concurrently with or after (subsequently or subsequent to) the administration of the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof.

[0271] In some embodiments, the dose of cells, or the subsequent dose of cells, is administered concurrently with initiating administration of the gamma secretase inhibitor in accord with the combination therapy methods. In some embodiments, the dose of cells, or the subsequent dose of cells, is administered on the same day as initiating administration of the gamma secretase inhibitor in accord with the combination therapy methods. In some embodiments, the dose of cells, or the subsequent dose of cells, is administered within 1 day, within 2 days, within 3 days, within 4 days, within 5 days, within 6 days, or within 7 days of initiating administration of the gamma secretase inhibitor in accord with the combination therapy methods.

[0272] In some embodiments, the dose of cells, or the subsequent dose of cells, is administered prior to starting or initiating administration of the gamma secretase inhibitor in accord with the provided combination therapy. In some embodiments, the dose of cells is administered at least or at least about 1 hour, at least or at least about 2 hours, at least or at least about 3 hours, at least or at least about 6 hours, at least or at least about 12 hours, at least or at least about 1 day, at least or at least about 2 days, at least or at least about 3 days, at least or about at least 4 days, at least or at least about 5 days, at least or about at least 6 days, at least or at least about 7 days, at least or about at least 12 days, at least or at least about 14 days, at least or about at least 15 days, at least or at least about 21 days, at least or at least about 28 days, at least or about at least 30 days, at least or at least about 35 days, at least or at least about 42 days, at least or about at least 60 days or at least or about at least 90 days prior to administering the gamma secretase inhibitor in accord with the provided combination therapy.

[0273] In some embodiments, the administration of the gamma secretase inhibitor in accord with the provided combination therapy is at a time in which the prior administration of the immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy) is associated with, or is likely to be associated with, a decreased functionality of the T cells compared to the functionality of the T cells at a time just prior to initiation of the immunotherapy (e.g., T cell therapy, such as CAR-T cell therapy) or at a preceding time point after initiation of the T cell therapy. In some embodiments, the method involves, subsequent to administering the dose of cells of the T cell therapy, e.g., adoptive T cell therapy, but prior to administering the gamma secretase inhibitor, assessing a sample from the subject for one or more functions of T cells, such as expansion or persistence of the cells, e.g. as determined by level or amount in the blood, or other phenotypes or desired outcomes as described herein, e.g., such as those described in Section III. Various parameters for determining or assessing the regimen of the combination therapy are described in Section III.

[0274] B. Administration of Gamma Secretase Inhibitor

[0275] In some embodiments of the methods, compositions, combinations, kits and uses provided herein, the combination therapy can be administered in one or more compositions, e.g., a pharmaceutical composition containing an inhibitor of gamma secretase, and/or the cell therapy, e.g., T cell therapy.

[0276] In some embodiments, the gamma secretase inhibitor inhibits or reduces the intramembrane cleavage of one or more receptors on a cell, for example, a tumor/cancer cell. In some embodiments, the IC.sub.50 of the inhibition of the intramembrane cleavage of the cell surface receptor on the cancer/tumor cell is less than about 100 .mu.M, 50 .mu.M, 10 .mu.M, 1 .mu.M, 0.75 .mu.M, 0.5 .mu.M, 0.25 .mu.M, 0.1 .mu.M, 75 nM, 50 nM, 25 nM, or 10 nM. In some embodiments, he gamma secretase inhibitor inhibits intramembrane cleavage of the cell surface receptor on the cancer/tumor cell with a half-maximal inhibitory concentration (IC.sub.50) of 0.01 nM to 10 nM, 0.01 nM to 5 nM, 0.01 nM to 1 nM, 0.01 nM, to 0.5 nM, 0.01 nM to 0.35 nM, 0.01 nM to 0.25 nM, 0.01 nM to 1.0 nM, 0.01 nM to 0.05 nM, 0.05 nM to 10 nM, 0.05 nM to 5 nM, 0.05 nM to 1 nM, 0.05 nM, to 0.5 nM, 0.05 nM to 0.35 nM, 0.05 nM to 0.25 nM, 0.05 nM to 1.0 nM, 1.0 nM to 10 nM, 1.0 nM nM to 5 nM, 1.0 nM nM to 1 nM, 1.0 nM nM, to 0.5 nM, 1.0 nM nM to 0.35 nM, 1.0 nM nM to 0.25 nM, 0.25 nM to 10 nM, 0.25 nM to 5 nM, 0.25 nM to 1 nM, 0.25 nM, to 0.5 nM, 0.25 nM to 0.35 nM, 0.35 nM to 10 nM, 0.35 nM to 5 nM, 0.35 nM to 1 nM, 0.35 nM, to 0.5 nM, 0.5 nM to 10 nM, 0.5 nM to 5 nM, 0.5 nM to 1 nM, 1 nM to 10 nM, 1 nM to 5 nM or 5 nM to 10 nM. In some embodiments, the IC.sub.50 of the inhibition of the intramembrane cleavage of the cell surface receptor on the cancer/tumor cell is about 10 nM-25 nM, 25 nM-50 nM, 50 nM-75 nM, 75 nM-0.1 .mu.M, 0.1 .mu.M-0.25 .mu.M, 0.25 .mu.M-0.5 .mu.M, 0.5 .mu.M-0.75 .mu.M, 0.75 .mu.M-1 .mu.M, 1 .mu.M-10 .mu.M, 10 .mu.M-25 .mu.M, 25 .mu.M-50 .mu.M, 50 .mu.M-75 .mu.M, or 75 .mu.M-100 .mu.M. In some embodiments, the receptor(s) is/are selected from the group consisting of B cell maturation antigen (BCMA), Notch 1, Notch 2, Notch 3, Notch 4, cell surface associated Mucin 1 (MUC1), Ephrin B2, Betaglycan (TGFBR3), CD43, CD44, CSF1R, CX3CR1, CXCL16, Delta1, E-cadherin, N-cadherin, HLA-A2, IFNaR2, IL1R1, IL1R2, IL6R, and amyloid precursor protein (APP). In some embodiments, the receptor is cell surface associated Mucin 1 (MUC1).

[0277] In some embodiments, the an inhibitor of gamma secretase inhibits or reduces the intramembrane cleavage of one or more receptors described above on a cell, for example, a tumor/cancer cell, and wherein the administration of the gamma secretase increases the level of the receptor expression on the cell. In some embodiments, the receptor is surface B cell maturation antigen (BCMA). In some embodiments, the gamma secretase inhibitor prohibits intramembrane cleavage of surface B cell maturation antigen (BCMA), and wherein the IC.sub.50 of the inhibition of the cleavage of surface BCMA is less than about 100 .mu.M, 50 .mu.M, 25 .mu., 10 .mu.M, 1 .mu.M, 0.75 .mu.M, 0.5 .mu.M, 0.25 .mu.M, 0.1 .mu.M, 75 nM, 50 nM, 25 nM, or 10 nM. In some embodiments, the IC.sub.50 of the inhibition of the intramembrane cleavage of the cell surface receptor on the cancer/tumor cell is about 10 nM-25 nM, 25 nM-50 nM, 50 nM-75 nM, 75 nM-0.1 .mu.M, 0.1 .mu.M-0.25 .mu.M, 0.25 .mu.M-0.5 .mu.M, 0.5 .mu.M-0.75 .mu.M, 0.75 .mu.M-1 .mu.M, 1 .mu.M-10 .mu.M, 10.mu.M-25 .mu.M, 25 .mu.M-50 .mu.M, 50 .mu.M-75 .mu.M, or 75 .mu.M-100 .mu.M. In some embodiments, the IC.sub.50 of the inhibition of the cleavage of surface BCMA is less than 0.5 nM. In some embodiments, the IC.sub.50 of the inhibition of the cleavage of surface BCMA is about 0.01 nM to about 0.5 nM, or 0.01 nM to about 0.5 nM, or about 0.01 nM to about 0.35 nM.

[0278] In some embodiments, the inhibitor of gamma secretase has a low brain penetration. In some embodiments, only 0.1%, 0.25%, 0.5%, 0.75%, 1%, 2.5%, 5%, 7.5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% of the gamma secretase inhibitor can penetrate the brain after administration.

[0279] In some embodiments, the inhibitor of gamma secretase selectively inhibits or reduces intramembrane cleavage of part of the substrates of gamma secretase. In some embodiments, the gamma secretase inhibitor selectively inhibits or reduces the intramembrane cleavage of surface B cell maturation antigen (BCMA).

[0280] In some embodiments, the gamma secretase inhibitor is a peptide inhibitor or non-peptide inhibitor. In some embodiments, the gamma secretase inhibitor is a peptide inhibitor from among peptide aldehydes derivatives, difluoroketones derivatives, hydroxyethylene dipeptide isotere derivatives, alpha-helical peptide derivatives and dipeptide analogs. In some embodiments, the gamma secretase inhibitor is a non-peptide inhibitor selected from benzodiazepines derivatives and sulfonamides derivatives. In some embodiments, the gamma secretase is a transition state inhibitor, for example, LY-4115754 or LY685,458. In some embodiments, the gamma secretase inhibitor is a non-transition state inhibitor such as DAPT, RO4929097. In some embodiments, the gamma secretase inhibitor is a gamma secretase modulator. In some embodiments, gamma secretase modulator is a nonsteroidal anti-inflammatory drugs-type modulator.

[0281] In some embodiments, the gamma secretase inhibitor is LY3039478, secretase inhibitor I (GSI I) Z-Leu-Leu-Norleucine; .gamma.-secretase inhibitor II (GSI II); .gamma.-secretase inhibitor III (GSI III), N-Benzyloxycarbonyl-Leu-leucinal, N-(2-Naphthoyl)-Val-phenylalaninal; .gamma.-secretase inhibitor III (GSI IV); .gamma.-secretase inhibitor III (GSI V), N-Benzyloxycarbonyl-Leu-phenylalaninal; .gamma.-secretase inhibitor III (GSI VI), 1-(S)-endo-N-(1,3,3)-Trimethylbicyclo[2.2.1]hept-2-yl)-4-fluorophenyl Sulfonamide; .gamma.-secretase inhibitor III (GSI VII), Menthyloxycarbonyl-LL-CHO; .gamma.-secretase inhibitor III (GSI IX), (DAPT), N-[N-(3,5-Difluorophenacetyl-L-alanyl)]-S-phenylglycine t-Butyl Ester; .gamma.-secretase inhibitor X (GSI X), {1S-Benzyl-4R-[1-(1S-carbamoyl-2-phenethylcarbamoyl)-1S-3-methylbutylcarb- -amoyl]-2R-hydroxy-5-phenylpentyl}carbamic Acid tert-butyl Ester; .gamma.-secretase inhibitor XI (GSI XI), 7-Amino-4-chloro-3-methoxyisocoumarin; .gamma.-secretase inhibitor XII (GSI XII), Z-Ile-Leu-CHO; .gamma.-secretase inhibitor XIII (GSI XIII), Z-Tyr-Ile-Leu-CHO; .gamma.-secretase inhibitor XIV (GSI XIV), Z-Cys(t-Bu)-Ile-Leu-CHO; .gamma.-secretase inhibitor XVI (GSI XVI), N-[N-3,5-Difluorophenacetyl]-L-alanyl-S-phenylglycine Methyl Ester; .gamma.-secretase inhibitor XVII (GSI XVII); .gamma.-secretase inhibitor XIX (GSI XIX), benzo[e][1,4]diazepin-3-yl)-butyramide; .gamma.-secretase inhibitor XX (GSI XX), (S,S)-2-[2-(3,5-Difluorophenyl)acetylamino]-N-(5-methyl-6-oxo-6,7-dihydro- -5H-dibenzo[b,d]azepin-7-yl)propionamide; .gamma.-secretase inhibitor XXI (GSI XXI), (S,S)-2-[2-(3,5-Difluorophenyl)-acetylamino]-N-(1-methyl-2-oxo-5-phenyl-2- -,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-propionamide; Gamma40 secretase inhibitor I, N-trans-3,5-Dimethoxycinnamoyl-Ile-leucinal; Gamma40 secretase inhibitor II, N-tert-Butyloxycarbonyl-Gly-Val-Valinal Isovaleryl-V V-Sta-A-Sta-OCH3; MK-0752 (Merck); MRK-003 (Merck); semagacestat/LY450139 (Eli Lilly); RO4929097; PF-03084,014; BMS-708163; MPC-7869 (.gamma.-secretase modifier), YO-01027 (Dibenzazepine), Compound E ([(2S)-2-{[(3,5-Difluorophenyl)acetyl]amino}-N-[(3S)-1-methyl-2-oxo-5-p- henyl-2,3-dihydro-1H-1,4-benzodiazepin-3-yl]propanamide], available from Alexis Biochemicals), LY411575 (Eli Lilly and Co.), L-685,458 (Sigma-Aldrich), BMS-289948 (4-chloro-N-(2,5-difluorophenyl)-N-((1R)-{4-fluoro-2-[3-(1H-imidazol-1-yl- )propyl]phenyl}ethyl)benzenesulfonamide hydrochloride) and BMS -299897 (4-[2-((1R)-1-{[(4-chlorophenyl)sulfonyl]-2,5-difluoroanilino}ethyl)-5-fl- uorophenyljbutanoic acid) (Bristol Myers Squibb).

[0282] In some embodiments, the gamma secretase inhibitor is LY3039478 or is a stereoisomer thereof or is a pharmaceutically acceptable salt or hydrate of the foregoing.

[0283] In some embodiments, the gamma secretase inhibitor is a compound of the structure:

##STR00009##

[0284] or a stereoisomer thereof or a pharmaceutically acceptable salt or hydrate of the foregoing.

[0285] In some embodiments, Compound 1 is a single stereoisomer as depicted. In some cases, there are two chiral centers giving rise to four stereoisomers. Thus, in some embodiments, reference to Compound 1 also includes racemic mixtures including Compound 1. Specific stereoisomers can be prepared by stereospecific synthesis using enantiomerically pure or enriched starting materials. The specific stereoisomers of either starting materials, intermediates, or racemic mixtures including Compound 1 can be resolved by techniques well known in the art, such as those found in Stereochemistry of Organic Compounds, E. I. Eliel and S. H. Wilen (Wiley 1994) and Enantiomers, Racemates, and Resolutions, J., Jacques, A. Collet, and S. H. Wilen (Wiley 1991), including chromatography on chiral stationary phases, enzymatic resolutions, or fractional crystallization or chromatography of diastereomers formed for that purpose, such as diastereomeric salts.

[0286] In some embodiments, the gamma secretase inhibitor is a compound of the structure:

##STR00010##

[0287] or a pharmaceutically acceptable salt or hydrate thereof.

[0288] In some embodiments, the gamma secretase inhibitor is a compound of the structure:

##STR00011##

[0289] or a pharmaceutically acceptable salt or hydrate thereof.

[0290] In some embodiments, the gamma secretase inhibitor is a compound of the structure:

##STR00012##

[0291] or a pharmaceutically acceptable salt or hydrate thereof.

[0292] In some embodiments, the gamma secretase inhibitor is a compound of the structure:

##STR00013##

[0293] or a pharmaceutically acceptable salt or hydrate thereof.

[0294] In some embodiments, the compound is Compound 1. In some embodiments, Compound 1 is named: 4,4,4-trifluoro-N-[(1S)-2-[[(7S)-5-(2-hydroxyethyl)-6-oxo-7H-pyrido[2,3-d- ][3]benzazepin-7-yl]amino]-1-methyl-2-oxo-ethyl]butanamide; and may also be named: N-[(1S)-2-[[(7S)-6,7-dihydro-5-(2-hydroxyethyl)-6-oxo-5H-pyrido- [3,2-a][3]benzazepin-7-yl]amino]-1-methyl-2-oxoethyl]-4,4,4-trifluorobutan- amide; and other names may be used to unambiguously identify Compound 1. In some embodiments, Compound 1 is known as LY3039478. See also published PCT App. No. WO2013/016081 for the synthesis and preparion of Compound 1.

[0295] 1. Compositions and Formulations

[0296] In some embodiments of the combination therapy methods, compositions, combinations, kits and uses provided herein, the combination therapy can be administered in one or more compositions, e.g., a pharmaceutical composition containing a gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof.

[0297] In some embodiments, the composition, e.g., a pharmaceutical composition containing the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, can include carriers such as a diluent, adjuvant, excipient, or vehicle with which gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, and/or the cells are administered. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin. Such compositions will contain a therapeutically effective amount of the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, generally in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. Saline solutions and aqueous dextrose and glycerol solutions also can be employed as liquid carriers, particularly for injectable solutions. The pharmaceutical compositions can contain any one or more of a diluents(s), adjuvant(s), antiadherent(s), binder(s), coating(s), filler(s), flavor(s), color(s), lubricant(s), glidant(s), preservative(s), detergent(s), sorbent(s), emulsifying agent(s), pharmaceutical excipient(s), pH buffering agent(s), or sweetener(s) and a combination thereof. In some embodiments, the pharmaceutical composition can be liquid, solid, a lyophilized powder, in gel form, and/or combination thereof. In some aspects, the choice of carrier is determined in part by the particular inhibitor and/or by the method of administration.

[0298] Pharmaceutically acceptable carriers are generally nontoxic to recipients at the dosages and concentrations employed, and include, but are not limited to: buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g. Zn-protein complexes); and/or non-ionic surfactants such as polyethylene glycol (PEG), stabilizers and/or preservatives. The compositions containing gamma secretase inhibitor or a pharmaceutically acceptable salt of hydrate thereof can also be lyophilized.

[0299] In some embodiments, the pharmaceutical compositions can be formulated for administration by any route known to those of skill in the art including intramuscular, intravenous, intradermal, intralesional, intraperitoneal injection, subcutaneous, intratumoral, epidural, nasal, oral, vaginal, rectal, topical, local, otic, inhalational, buccal (e.g., sublingual), and transdermal administration or any route. In some embodiments, other modes of administration also are contemplated. In some embodiments, the administration is by bolus infusion, by injection, e.g., intravenous or subcutaneous injections, intraocular injection, periocular injection, subretinal injection, intravitreal injection, trans-septal injection, subscleral injection, intrachoroidal injection, intracameral injection, subconjectval injection, subconjuntival injection, sub-Tenon's injection, retrobulbar injection, peribulbar injection, or posterior juxtascleral delivery. In some embodiments, administration is by parenteral, intrapulmonary, and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. In some embodiments, a given dose is administered by a single bolus administration. In some embodiments, it is administered by multiple bolus administrations, for example, over a period of no more than 3 days, or by continuous infusion administration.

[0300] In some embodiments, the administration can be local, topical or systemic depending upon the locus of treatment. In some embodiments local administration to an area in need of treatment can be achieved by, for example, but not limited to, local infusion during surgery, topical application, e.g., in conjunction with a wound dressing after surgery, by injection, by means of a catheter, by means of a suppository, or by means of an implant. In some embodiments, compositions also can be administered with other biologically active agents, either sequentially, intermittently or in the same composition. In some embodiments, administration also can include controlled release systems including controlled release formulations and device controlled release, such as by means of a pump. In some embodiments, the administration is oral.

[0301] In some embodiments, pharmaceutically and therapeutically active compounds and derivatives thereof are typically formulated and administered in unit dosage forms or multiple dosage forms. Each unit dose contains a predetermined quantity of therapeutically active compound sufficient to produce the desired therapeutic effect, in association with the required pharmaceutical carrier, vehicle or diluent. In some embodiments, unit dosage forms, include, but are not limited to, tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil water emulsions containing suitable quantities of the compounds or pharmaceutically acceptable derivatives thereof. Unit dose forms can be contained ampoules and syringes or individually packaged tablets or capsules. Unit dose forms can be administered in fractions or multiples thereof. In some embodiments, a multiple dose form is a plurality of identical unit dosage forms packaged in a single container to be administered in segregated unit dose form. Examples of multiple dose forms include vials, bottles of tablets or capsules or bottles of pints or gallons.

[0302] 2. Dosage Schedule of Gamma Secretase Inhibitor

[0303] In some embodiments, the provided combination therapy method involves administering to the subject a therapeutically effective amount of a gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, and the cell therapy, such as a T cell therapy (e.g. CAR-expressing T cells).

[0304] In some embodiments, the administration of the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, is initiated prior to, subsequently to, during, during the course of, simultaneously, near simultaneously, sequentially and/or intermittently with the administration of the cell therapy, such as a T cell therapy (e.g. CAR-expressing T cells). In some embodiments, the method involves initiating the administration of gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, prior to administration of the T cell therapy. In other embodiments, the method involves initiating the administration of the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, after administration of a cell therapy (e.g., a CAR-expressing T cell therapy). In some embodiments, the dosage schedule comprises initiating the administration of gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, concurrently or simultaneously with the administration of a cell therapy (e.g., a CAR-expressing T cell therapy).

[0305] In some embodiments, the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof is administered in a cycle. In some embodiments, the cycle comprises an administration period in which the gamma secretase inhibitor is administered followed by a rest period during which the gamma secretase inhibitor is not administered. In some embodiments, the total number of days of the cycle, e.g. from the beginning of initiating administration of the gamma secretase inhibitor, is greater than or greater than about or is about 1 day, 3, days, 7 days, 14 days, 21 days, 28 days, 30 days, 40 days, 50 days, 60 days or more.

[0306] In some embodiments, the initiation of the administration of the gamma secretase inhibitor is carried out in at least one cycle and initiation of administration of the cell therapy (e.g., CAR-expressing T cells) are carried out on the same day, optionally concurrently. In some embodiments, the initiation of the administration of the gamma secretase inhibitor in at least one cycle is prior to initiation of administration of the cell therapy. In some embodiments, the initiation of the administration of the gamma secretase inhibitor in at least one cycle is concurrent with or on the same day as initiation of administration of the cell therapy. In some embodiments, the gamma secretase inhibitor is administered from or from about 0 to 30 days, such as 0 to 15 days, 0 to 6 days, 0 to 96 hours, 0 to 24 hours, 0 to 12 hours, 0 to 6 hours, or 0 to 2 hours, 2 hours to 15 days, 2 hours to 6 days, 2 hours to 96 hours, 2 hours to 24 hours, 2 hours to 12 hours, 2 hours to 6 hours, 6 hours to 30 days, 6 hours to 15 days, 6 hours to 6 days, 6 hours to 96 hours, 6 hours to 24 hours, 6 hours to 12 hours, 12 hours to 30 days, 12 hours to 15 days, 12 hours to 6 days, 12 hours to 96 hours, 12 hours to 24 hours, 24 hours to 30 days, 24 hours to 15 days, 24 hours to 6 days, 24 hours to 96 hours, 96 hours to 30 days, 96 hours to 15 days, 96 hours to 6 days, 6 days to 30 days, 6 days to 15 days, or 15 days to 30 days prior to initiation of the cell therapy (e.g., CAR-expressing T cell therapy). In some aspects, the gamma secretase inhibitor is administered no more than about 96 hours, 72 hours, 48 hours, 24 hours, 12 hours, 6 hours, 2 hours or 1 hour prior to initiation of the T cell therapy.

[0307] In some of any such embodiments in which the gamma secretase inhibitor, or a pharmaceutically acceptable salt of hydrate thereof, is given prior to the cell therapy (e.g. T cell therapy, such as CAR-T cell therapy), the administration of the gamma secretase inhibitor, continues at regular intervals until the initiation of the cell therapy and/or for a time after the initiation of the cell therapy.

[0308] In some embodiments, the gamma secretase inhibitor is administered, or is further administered, after administration of the cell therapy (e.g. T cell therapy, such as CAR-T cell therapy). In some embodiments, the gamma secretase inhibitor is administered within or within about 1 hours, 2 hours, 6 hours, 12 hours, 24 hours, 48 hours, 96 hours, 4 days, 5 days, 6 days or 7 days, 14 days, 15 days, 21 days, 24 days, 28 days, 30 days, 36 days, 42 days, 60 days, 72 days or 90 days after initiation of administration of the cell therapy (e.g. T cell therapy).

[0309] In some embodiments, the provided methods involve continued administration, such as at regular intervals, of the gamma secretase inhibitor after initiation of administration of the cell therapy. For example, the gamma secretase inhibitor is administered at regular intervals of daily, every other day, every third day, three times a week, or once a week for a period of time.

[0310] In some embodiments, the gamma secretase inhibitor is administered, such as at regular intervals as described, for up to or up to about 1 day, up to or up to about 2 days, up to or up to about 3 days, up to or up to about 4 days, up to or up to about 5 days, up to or up to about 6 days, up to or up to about 7 days, up to or up to about 12 days, up to or up to about 14 days, up to or up to about 21 days, up to or up to about 24 days, up to or up to about 28 days, up to or up to about 30 days, up to or up to about 35 days, up to or up to about 42 days, up to or up to about 60 days or up to or up to about 90 days, up to or up to about 120 days, up to or up to about 180 days, up to or up to about 240 days, up to or up about 360 days, or up to or up to about 720 days or more after the initiation of administration of the cell therapy (e.g. T cell therapy, such as CAR-T cell therapy).

[0311] In some of any such above embodiments, the gamma secretase inhibitor is administered prior to and after initiation of administration of the cell therapy (e.g. T cell therapy, such as CAR-T cell therapy).

[0312] In some embodiments, the initiation of the administration of the gamma secretase inhibitor is carried out at or after, optionally immediately after or within 1 to 3 days after: (i) peak or maximum level of the cells of the T cell therapy are detectable in the blood of the subject; (ii) the number of cells of the T cell therapy detectable in the blood, after having been detectable in the blood, is not detectable or is reduced, optionally reduced compared to a preceding time point after administration of the T cell therapy; (iii) the number of cells of the T cell therapy detectable in the blood is decreased by or more than 1.2-fold, 1.5-fold, 2.0-fold, 3.0-fold, 4.0-fold, 5.0-fold, 10-fold or more the peak or maximum number cells of the T cell therapy detectable in the blood of the subject after initiation of administration of the T cell therapy; (iv) at a time after a peak or maximum level of the cells of the T cell therapy are detectable in the blood of the subject, the number of cells of or derived from the T cells detectable in the blood from the subject is less than 10%, less than 5%, less than 1% or less than 0.1% of total peripheral blood mononuclear cells (PBMCs) in the blood of the subject; (v) the subject exhibits disease progression and/or has relapsed following remission after treatment with the T cell therapy; and/or (iv) the subject exhibits increased tumor burden as compared to tumor burden at a time prior to or after administration of the T cells and prior to initiation of administration of the gamma secretase inhibitor.

[0313] In some aspects, initiation of administration of the gamma secretase inhibitor is at a time after the subject has relapsed or is suspected to or likely to relapse following administration of the cell therapy (e.g. T cell therapy, such as CAR T-cell therapy). In some cases, the cell therapy is an anti-BCMA CAR-T cell therapy and the gamma secretase inhibitor is administered at a time after the subject has relapsed or is suspected to or likely to relapse following administration of the anti-BCMA CAR-T cell therapy.

[0314] In some embodiments, the initiation of the administration of the gamma secretase inhibitor in at least one cycle is after initiation of administration of the cell therapy. In some embodiments, the initiation of the administration of the gamma secretase inhibitor is at least or about at least 1 day, at least or about at least 2 days, at least or about at least 3 days, at least or about at least 4 days, at least or about at least 5 days, at least or about at least 6 days, at least or about at least 7 days, at least or about at least 8 days, at least or about at least 9 days, at least or about at least 10 days, at least or at least about 12 days, at least or about at least 14 days, at least or at least about 15 days, at least or about at least 21 days, at least or at least about 24 days, at least or about at least 28 days, at least or about at least 30 days, at least or about at least 35 days or at least or about at least 42 days, at least or about at least 60 days, or at least or about at least 90 days after initiation of the administration of the cell therapy. In some embodiments, the initiation of the administration of the gamma secretase inhibitor is carried out at least 2 days after, at least 1 week after, at least 2 weeks after, at least 3 weeks after, or at least 4 weeks after, the initiation of the administration of the cell therapy. In some embodiments, the initiation of the administration of the gamma secretase inhibitor is carried out 2 to 28 days or 7 to 21 days after initiation of administration of the cell therapy. In some embodiments, the initiation of the administration of the gamma secretase inhibitor is carried out at a time that is greater than or greater than about 14 days, 15 days, 16 days, 17 days, 18 days, 19, days, 20 days, 21 days, 24 days, or 28 days after initiation of the administration of the cell therapy.

[0315] In some embodiments, the gamma secretase inhibitor is administered several times a day, twice a day, daily, every other day, three times a week, twice a week, or once a week after initiation of the cell therapy. In some embodiments, the gamma secretase inhibitor is administered daily. In some embodiments the gamma secretase inhibitor is administered twice a day. In some embodiments, the gamma secretase inhibitor is administered three times a day. In other embodiments, the gamma secretase inhibitor is administered every other day.

[0316] In some embodiments, the gamma secretase inhibitor is administered daily. In some embodiments, the gamma secretase inhibitor is administered during the administration period for a plurality of consecutive days, such as for up to about 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more than 30 consecutive days. In some embodiments, the gamma secretase inhibitor, is administered for greater than or greater than about 7 consecutive days, greater than or greater than about 14 consecutive days, greater than or greater than about 21 consecutive days, greater than or greater than about 21 consecutive days, or greater than or greater than about 28 consecutive days. In some embodiments, the gamma secretase inhibitor, is administered during the administration period for up to 21 consecutive days. In some embodiments, the gamma secretase inhibitor, is administered during the administration period for up to 21 consecutive days, wherein the cycle comprises greater than 30 days beginning upon initiation of the administration of the gamma secretase inhibitor.

[0317] In some embodiments, the gamma secretase inhibitor is administered during the administration period for no more than about 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or no more than 30 consecutive days. In certain embodiments, the gamma secretase inhibitor is administered once daily for 14 days over a 21 day treatment cycle. In certain embodiments, the gamma secretase inhibitor is administered once daily for 21 days over a 28 day treatment cycle. In some embodiments, the gamma secretase inhibitor is administered during the administration period for no more than 14 consecutive days.

[0318] In some embodiments, the gamma secretase inhibitor is administered in a cycle, wherein the cycle comprises the administration of the gamma secretase inhibitor for a plurality of consecutive days followed by a rest period during which the gamma secretase inhibitor is not administered. In some embodiments, the rest period is greater than about 1 day, greater than about 3 consecutive days, greater than about 5 consecutive days, greater than about 7 consecutive days, greater than about 8 consecutive days, greater than about 9 consecutive days, greater than about 10 consecutive days, greater than about 11 consecutive days, greater than about 12 consecutive days, greater than about 13 consecutive days, greater than about 14 consecutive days, greater than about 15 consecutive days, greater than about 16 consecutive days, greater than about 17 consecutive days, greater than about 18 consecutive days, greater than about 19 consecutive days, greater than about 20 consecutive days, or greater than about 21 or more consecutive days. In some embodiments, the rest period is greater than 7 consecutive days, greater than 14 consecutive days, greater than 21 days, or greater than 28 days. In some embodiments, the rest period is greater than about 14 consecutive days. In some embodiments, the cycle of administration of the gamma secretase inhibitor does not contain a rest period.

[0319] In some embodiments, the gamma secretase inhibitor is administered in a cycle, wherein the cycle is repeated at least one time. In some embodiments, the gamma secretase inhibitor is administered for at least 2 cycles, at least 3 cycles, at least 4 cycles, at least 5 cycles, at least 6 cycles, at least 7 cycles, at least 8 cycles, at least 9 cycles, at least 10 cycles, at least 11 cycles, or at least 12 cycles. In some embodiments, the gamma secretase inhibitor is administered for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24 cycles.

[0320] In some embodiments, the gamma secretase inhibitor is administered at least six times daily, five times daily, four times daily, three times daily, twice daily, once daily, every other day, every three days, three times a week, twice weekly, once weekly or only one time prior to or subsequently to initiation of administration of the cell therapy. In some embodiments, the gamma secretase inhibitor is administered three times a week. In some embodiments, the administration of the inhibitor is carried out in a treatment cycle that is at least or at least about or 14 days, at least or at least about or 21 days or at least or at least about or 28 days. In some embodiments, the gamma secretase inhibitor is administered in multiple doses in regular intervals prior to, during, during the course of, and/or after the period of administration of the cell therapy. In some embodiments, the gamma secretase inhibitor is administered in one or more doses in regular intervals prior to the administration of the cell therapy. In some embodiments, the gamma secretase inhibitor is administered in one or more doses in regular intervals after the administration of the cell therapy. In some embodiments, one or more of the doses of the gamma secretase inhibitor, can occur simultaneously with the administration of a dose of the cell therapy.

[0321] In some embodiments, the dose, frequency, duration, timing and/or order of administration of the gamma secretase inhibitor, is determined, based on particular thresholds or criteria of results of the screening step and/or assessment of treatment outcomes described herein, e.g., those described in Section III herein.

[0322] In some embodiments, the method involves administering the cell therapy to a subject that has been previously administered a therapeutically effective amount of the gamma secretase inhibitor. In some embodiments, the gamma secretase inhibitor is administered to a subject before administering a dose of cells expressing a recombinant receptor to the subject. In some embodiments, the treatment with the gamma secretase inhibitor occurs at the same time as the administration of the dose of cells. In some embodiments, the gamma secretase inhibitor is administered after the administration of the dose of cells.

[0323] In some embodiments, administration of the gamma secretase inhibitor is carried out for an administration period of 2 to 28 days, such as 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days. In some embodiments, the gamma secretase inhibitor is administered daily for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days. In some embodiments, the gamma secretase inhibitor is administered twice a day for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days. In some embodiments, the gamma secretase inhibitor is administered three times a day for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days. In some embodiments, the gamma secretase inhibitor is administered every other day for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days. In some embodiments, the gamma secretase inhibitor is administered three times a week for 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days.

[0324] In some embodiments, the gamma secretase inhibitor is administered on days 2, 4, 7, 9, 11, 14, 16, and 18 after initiation of administration of the cell therapy (e.g. T cell therapy).

[0325] In some embodiments of the methods provided herein, the gamma secretase inhibitor, and the cell therapy are administered simultaneously or near simultaneously.

[0326] In some embodiments, gamma secretase inhibitor is administered at a total daily dosage amount of at most or at most about 50 mg/day, 100 mg/day, 150 mg/day, 175 mg/day, 200 mg/day, 250 mg/day, 300 mg/day, 350 mg/day, 400 mg/day, 450 mg/day, 500 mg/day, 600 mg/day, 700 mg/day, 800 mg/day or less. In some embodiments, the gamma secretase inhibitor is administered, or each administration of the inhibitor is independently administered, in an amount of 0.5 mg to 500 mg, 0.5 mg to 250 mg, 0.5 mg to 100 mg, 0.5 mg to 50 mg, 0.5 mg to 25 mg, 0.5 mg to 10 mg, 0.5 mg to 5.0 mg, 0.5 mg to 2.5 mg, 0.5 mg to 1.0 mg, 1.0 mg to 500 mg, 1.0 mg to 250 mg, 1.0 mg to 100 mg, 1.0 mg to 50 mg, 1.0 mg to 25 mg, 1.0 mg to 10 mg, 1.0 mg to 5.0 mg, 1.0 mg to 2.5 mg, 2.5 mg to 500 mg, 2.5 mg to 250 mg, 2.5 mg to 100 mg, 2.5 mg to 50 mg, 2.5 mg to 25 mg, 2.5 mg to 10 mg, 2.5 mg to 5.0 mg, 5.0 mg to 500 mg, 5.0 mg to 250 mg, 5.0 mg to 100 mg, 5.0 mg to 50 mg, 5.0 mg to 25 mg, 5.0 mg to 10 mg, 10 mg to 500 mg, 10 mg to 250 mg, 10 mg to 100 mg, 10 mg to 50 mg, 10 mg to 25 mg, 25 mg to 500 mg, 25 mg to 250 mg, 25 mg to 100 mg, 25 mg to 50 mg, 50 mg to 500 mg, 50 mg to 250 mg, 50 mg to 100 mg, 100 mg to 500 mg, 100 mg to 250 mg or 250 mg to 500 mg. In some embodiments, the inhibitor is administered, or each administration of the inhibitor is independently administered, in an amount that is at least or at least about or is or is about 0.5 mg, 1.0 mg, 2.5 mg, 5.0 mg, 10.0 mg, 25 mg, 50 mg, 100 mg, 250 mg or 500 mg. In some embodiments, the amount is a once daily amount of the gamma secretase inhibitor. Such dosage amounts can be administered at regular intervals as described, such as every other day, every third day, three times a week or once a week for an administration period. In some embodiments, the administration period is 2 to 28 days, such as 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days.

[0327] In some embodiments, the gamma secretase inhibitor is administered at a dosage of from about 1 mg to about 20 mg, e.g., from about 1 mg to about 10 mg, from about 2.5 mg to about 7.5 mg, from about 5 mg to about 15 mg, such as about 5 mg, 10 mg, 15 mg or 20 mg. In some embodiments, the gamma secretase inhibitor is administered at a dose of from at or about 10 .mu.g/kg to at or about 5 mg/kg, such as at or about 50 .mu.g/kg to at or about 2 mg/kg , at or about 50 .mu.g/kg to at or about 1 mg/kg, at or about 50 .mu.g/kg to at or about 500 .mu.g/kg, at or about 50 .mu.g/kg to at or about 250 mg/kg, at or about 100 .mu.g/kg to at or about 200 mg/kg, at or about 50 to at or bout 100 .mu.g/kg, at or about 100 .mu.g/kg to about 2 mg/kg, at or about 100 .mu.g/kg to at or about 1 mg/kg, at or about 100 .mu.g/kg to at or about 500 .mu.g/kg, at or about 100 .mu.g/kg to at or about 250 .mu.g/kg, at or about 100 .mu.g/kg to at or about 200 .mu.g/kg, at or about 100 .mu.g/kg, at or about 200 .mu.g/kg to about 2 mg/kg, at or about 200 .mu.g/kg to at or about 1 mg/kg, at or about 200 .mu.g/kg to at or about at or about 500 mg/kg, at or about 200 .mu.g/kg to at or about 250 .mu.g/kg or at or about 250 .mu.g/kg to at or about 2 mg/kg, at or about 250 .mu.g/kg to at or about 1 mg/kg, at or about 250 .mu.g/kg to at or about at or about 500 .mu.g/kg, In some embodiments, the gamma secretase inhibitor is administered at a dose of from at or about 400 .mu.g/kg to at or about 600 .mu.g/kg, such as at or about 500 .mu.g/kg. In some embodiments, the amount is a once daily amount of the gamma secretase inhibitor. Such dosage amounts can be administered at regular intervals as described, such as every other day, every third day, three times a week or once a week for an administration period. In some embodiments, the administration period is 2 to 28 days, such as 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days.

[0328] In some embodiments, the gamma secretase inhibitor is administered at a total daily dosage amount of at least or at least about 0.1 mg per day, 0.5 mg per day, 1.0 mg per day, 2.5 mg per day, 5 mg per day, 10 mg per day, 25 mg per day, 50 mg per day or 100 mg per day. In some embodiments, the dose of the inhibitor is about 25 mg per day. In particular embodiments, the dose of the inhibitor is or is about10 mg per day. Such dosage amounts can be administered at regular intervals as described, such as every other day, every third day, three times a week or once a week for an administration period. In some embodiments, the administration period is 2 to 28 days, such as 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days.

[0329] In some embodiments, the gamma secretase inhibitor is administered in an amount greater than or greater than about 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg and less than 25 mg. In some embodiments, the gamma secretase inhibitor is administered in an amount greater than or greater than about 1 mg per day, 2.5 mg per day, 5 mg per day, 7.5 mg per day, 10 mg per day, 15 mg per day and less than 25 mg per day. Such dosage amounts can be administered at regular intervals as described, such as every other day, every third day, three times a week or once a week for an administration period. In some embodiments, the administration period is 2 to 28 days, such as 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or more than 21 days.

[0330] In some embodiments, the gamma secretase inhibitor is administered on days 2, 4, 7, 9, 11, 14, 16, and 18 after initiation of administration of the cell therapy (e.g. T cell therapy).

[0331] In some embodiments, dosages, such as daily dosages, are administered in one or more divided doses, such as 2, 3, or 4 doses, or in a single formulation. The gamma secretase inhibitor can be administered alone, in the presence of a pharmaceutically acceptable carrier, or in the presence of other therapeutic agents.

[0332] One skilled in the art will recognize that higher or lower dosages of the gamma secretase inhibitor could be used, for example depending on the particular agent and the route of administration. In some embodiments, the gamma secretase inhibitor may be administered alone or in the form of a pharmaceutical composition wherein the compound is in admixture or mixture with one or more pharmaceutically acceptable carriers, excipients, or diluents. In some embodiments, the gamma secretase inhibitor may be administered either systemically or locally to the organ or tissue to be treated. Exemplary routes of administration include, but are not limited to, topical, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, intratumoral, and intravenous), oral, sublingual, rectal, transdermal, intranasal, vaginal and inhalation routes. In some embodiments, the route of administration is oral, parenteral, rectal, nasal, topical, or ocular routes, or by inhalation. In some embodiments, the gamma secretase inhibitor is administered orally. In some embodiments, the gamma secretase inhibitor is administered orally in solid dosage forms, such as capsules, tablets and powders, or in liquid dosage forms, such as elixirs, syrups and suspensions.

[0333] Once improvement of the patient's disease has occurred, the dose may be adjusted for preventative or maintenance treatment. For example, the dosage or the frequency of administration, or both, may be reduced as a function of the symptoms, to a level at which the desired therapeutic or prophylactic effect is maintained. If symptoms have been alleviated to an appropriate level, treatment may cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms. Patients may also require chronic treatment on a long-term basis.

[0334] 3. Additional Therapy

[0335] In some aspects, the provided methods can further include administering one or more lymphodepleting therapies, such as prior to or simultaneous with initiation of administration of the cell therapy (e.g., T cell therapy). In some embodiments, the lymphodepleting therapy comprises administration of a phosphamide, such as cyclophosphamide. In some embodiments, the lymphodepleting therapy can include administration of fludarabine.

[0336] In some aspects, preconditioning subjects with immunodepleting (e.g., lymphodepleting) therapies can improve the effects of adoptive cell therapy (ACT). Preconditioning with lymphodepleting agents, including combinations of cyclosporine and fludarabine, have been effective in improving the efficacy of transferred tumor infiltrating lymphocyte (TIL) cells in cell therapy, including to improve response and/or persistence of the transferred cells. See, e.g., Dudley et al., Science, 298, 850-54 (2002); Rosenberg et al., Clin Cancer Res, 17(13):4550-4557 (2011). Likewise, in the context of CAR+ T cells, several studies have incorporated lymphodepleting agents, most commonly cyclophosphamide, fludarabine, bendamustine, or combinations thereof, sometimes accompanied by low-dose irradiation. See Han et al. Journal of Hematology & Oncology, 6:47 (2013); Kochenderfer et al., Blood, 119: 2709-2720 (2012); Kalos et al., Sci Transl Med, 3(95):95ra73 (2011); Clinical Trial Study Record Nos.: NCT02315612; NCT01822652.

[0337] Such preconditioning can be carried out with the goal of reducing the risk of one or more of various outcomes that could dampen efficacy of the therapy. These include the phenomenon known as "cytokine sink," by which T cells, B cells, NK cells compete with TILs for homeostatic and activating cytokines, such as IL-2, IL-7, and/or IL-15; suppression of TILs by regulatory T cells, NK cells, or other cells of the immune system; impact of negative regulators in the tumor microenvironment. Muranski et al., Nat Clin Pract Oncol. December; 3(12): 668-681 (2006).

[0338] Thus in some embodiments, the provided method further involves administering a lymphodepleting therapy to the subject. In some embodiments, the method involves administering the lymphodepleting therapy to the subject prior to the administration of the dose of cells. In some embodiments, the lymphodepleting therapy contains a chemotherapeutic agent such as fludarabine and/or cyclophosphamide. In some embodiments, the administration of the cells and/or the lymphodepleting therapy is carried out via outpatient delivery.

[0339] In some embodiments, the methods include administering a preconditioning agent, such as a lymphodepleting or chemotherapeutic agent, such as cyclophosphamide, fludarabine, or combinations thereof, to a subject prior to the administration of the dose of cells. For example, the subject may be administered a preconditioning agent at least 2 days prior, such as at least 3, 4, 5, 6, or 7 days prior, to the first or subsequent dose. In some embodiments, the subject is administered a preconditioning agent no more than 7 days prior, such as no more than 6, 5, 4, 3, or 2 days prior, to the administration of the dose of cells.

[0340] In some embodiments, the subject is preconditioned with cyclophosphamide at a dose between or between about 20 mg/kg and 100 mg/kg of body surface area of the subject, such as between or between about 40 mg/kg and 80 mg/kg. In some aspects, the subject is preconditioned with or with about 60 mg/kg of cyclophosphamide. In some embodiments, the fludarabine can be administered in a single dose or can be administered in a plurality of doses, such as given daily, every other day or every three days. In some embodiments, the cyclophosphamide is administered once daily for one or two days. In some embodiments, where the lymphodepleting agent comprises cyclophosphamide, the subject is administered cyclophosphamide at a dose between or between about 100 mg/m.sup.2 and 500 mg/m.sup.2 body surface area of the subject, such as between or between about 200 mg/m.sup.2 and 400 mg/m.sup.2, or 250 mg/m.sup.2 and 350 mg/m.sup.2, inclusive. In some instances, the subject is administered about 300 mg/m.sup.2 of cyclophosphamide. In some embodiments, the cyclophosphamide can be administered in a single dose or can be administered in a plurality of doses, such as given daily, every other day or every three days. In some embodiments, cyclophosphamide is administered daily, such as for 1-5 days, for example, for 2 to 4 days. In some instances, the subject is administered about 300 mg/m.sup.2 body surface area of the subject, of cyclophosphamide, daily for 3 days, prior to initiation of the cell therapy. In some embodiments, where the lymphodepleting agent comprises fludarabine, the subject is administered fludarabine at a dose between or between about 1 mg/m.sup.2 and 100 mg/m.sup.2 of body surface area of the subject, such as between or between about 10 mg/m.sup.2 and 75 mg/m.sup.2, 15 mg/m.sup.2 and 50 mg/m.sup.2, 20 mg/m.sup.2 and 30 mg/m.sup.2, 20 mg/m.sup.2 and 40 mg/m.sup.2, 24 mg/m.sup.2 and 35 mg/m.sup.2, or 24 mg/m.sup.2 and 26 mg/m.sup.2, each inclusive. In some instances, the subject is administered about 30 mg/m.sup.2 of fludarabine. In some instances, the subject is administered 25 mg/m.sup.2 of fludarabine. In some embodiments, the fludarabine can be administered in a single dose or can be administered in a plurality of doses, such as given daily, every other day or every three days. In some embodiments, fludarabine is administered daily, such as for 1-5 days, for example, for 3 to 5 days or for 3-4 days. In some instances, the subject is administered about 30 mg/m.sup.2 body surface area of the subject, of fludarabine, daily for 3 days, prior to initiation of the cell therapy.

[0341] In some embodiments, the lymphodepleting agent comprises a combination of agents, such as a combination of cyclophosphamide and fludarabine. Thus, the combination of agents may include cyclophosphamide at any dose or administration schedule, such as those described above, and fludarabine at any dose or administration schedule, such as those described above. For example, in some aspects, the subject is administered 60 mg/kg (-2 g/m.sup.2) of cyclophosphamide and 3 to 5 doses of 25 mg/m.sup.2 fludarabine prior to the dose of cells. In some aspects, the subject is administered fludarabine at or about 30 mg/m.sup.2 body surface area of the subject, daily, and cyclophosphamide at or about 300 mg/m.sup.2 body surface area of the subject, daily, for 3 days.

[0342] In one exemplary dosage regime, prior to receiving the first dose, subjects receive a gamma secretase inhibitor 1 day before the administration of cells and a lymphodepleting preconditioning chemotherapy of cyclophosphamide and fludarabine (CY/FLU), which is administered at least two days before the first dose of CAR-expressing cells and generally no more than 7 days before administration of cells. In another exemplary dosage regime, subjects receive the gamma secretase inhibitor concurrently with the administration of cells, such as on the same day. In yet another exemplary dosage regime, subjects receive the gamma secretase inhibitor several days after the administration of cells, such as 7, 8, 9, 10, 11, 12, 13, 14, or more than 14 days after. In some cases, for example, cyclophosphadmide is given from 24 to 27 days after the administration of the gamma secretase inhibitor. After preconditioning treatment, subjects are administered the dose of CAR-expressing T cells as described above.

[0343] In some embodiments, the administration of the preconditioning agent prior to infusion of the dose of cells improves an outcome of the treatment. For example, in some aspects, preconditioning improves the efficacy of treatment with the dose or increases the persistence of the recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells) in the subject. In some embodiments, preconditioning treatment increases disease-free survival, such as the percent of subjects that are alive and exhibit no minimal residual or molecularly detectable disease after a given period of time following the dose of cells. In some embodiments, the time to median disease-free survival is increased.

[0344] Once the cells are administered to the subject (e.g., human), the biological activity of the engineered cell populations in some aspects is measured by any of a number of known methods. Parameters to assess include specific binding of an engineered or natural T cell or other immune cell to antigen, in vivo, e.g., by imaging, or ex vivo, e.g., by ELISA or flow cytometry. In certain embodiments, the ability of the engineered cells to destroy target cells can be measured using any suitable method known in the art, such as cytotoxicity assays described in, for example, Kochenderfer et al., J. Immunotherapy, 32(7): 689-702 (2009), and Herman et al. J. Immunological Methods, 285(1): 25-40 (2004). In certain embodiments, the biological activity of the cells also can be measured by assaying expression and/or secretion of certain cytokines, such as CD 107a, IFN.gamma., IL-2, and TNF. In some aspects the biological activity is measured by assessing clinical outcome, such as reduction in tumor burden or load. In some aspects, toxic outcomes, persistence and/or expansion of the cells, and/or presence or absence of a host immune response, are assessed.

[0345] In some embodiments, the administration of the preconditioning agent prior to infusion of the dose of cells improves an outcome of the treatment such as by improving the efficacy of treatment with the dose or increases the persistence of the recombinant receptor-expressing cells (e.g., CAR-expressing cells, such as CAR-expressing T cells) in the subject. Therefore, in some embodiments, the dose of preconditioning agent given in the method which is a combination therapy with the gamma secretase inhibitor and cell therapy is higher than the dose given in the method without the gamma secretase inhibitor.

II. CELL THERAPY AND ENGINEERING CELLS

[0346] In some embodiments, the cell therapy (e.g., T cell therapy) for use in accord with the provided combination therapy methods includes administering engineered cells expressing recombinant receptors designed to recognize and/or specifically bind to molecules associated with the disease or condition and result in a response, such as an immune response against such molecules upon binding to such molecules. The receptors may include chimeric receptors, e.g., chimeric antigen receptors (CARs), and other transgenic antigen receptors including transgenic T cell receptors (TCRs).

[0347] In some embodiments, the cells contain or are engineered to contain an engineered receptor, e.g., an engineered antigen receptor, such as a chimeric antigen receptor (CAR), or a T cell receptor (TCR). Also provided are populations of such cells, compositions containing such cells and/or enriched for such cells, such as in which cells of a certain type such as T cells or CD8.sup.+ or CD4.sup.+ cells are enriched or selected. Among the compositions are pharmaceutical compositions and formulations for administration, such as for adoptive cell therapy. Also provided are therapeutic methods for administering the cells and compositions to subjects, e.g., patients.

[0348] Thus, in some embodiments, the cells include one or more nucleic acids introduced via genetic engineering, and thereby express recombinant or genetically engineered products of such nucleic acids. In some embodiments, gene transfer is accomplished by first stimulating the cells, such as by combining it with a stimulus that induces a response such as proliferation, survival, and/or activation, e.g., as measured by expression of a cytokine or activation marker, followed by transduction of the activated cells, and expansion in culture to numbers sufficient for clinical applications.

[0349] A. Recombinant Receptors

[0350] The cells generally express recombinant receptors, such as antigen receptors including functional non-TCR antigen receptors, e.g., chimeric antigen receptors (CARs), and other antigen-binding receptors such as transgenic T cell receptors (TCRs). Also among the receptors are other chimeric receptors.

[0351] In some embodiments of the provided methods and uses, recombinant receptors, including chimeric receptors, such as a chimeric antigen receptors, contain one or more domains that combine a ligand-binding domain (e.g. antibody or antibody fragment) that provides specificity for a desired antigen (e.g., tumor antigen) with intracellular signaling domains. In some embodiments, the intracellular signaling domain is an activating intracellular domain portion, such as a T cell activating domain, providing a primary activation signal. In some embodiments, the intracellular signaling domain contains or additionally contains a costimulatory signaling domain to facilitate effector functions. In some embodiments, chimeric receptors when genetically engineered into immune cells can modulate T cell activity, and, in some cases, can modulate T cell differentiation or homeostasis, thereby resulting in genetically engineered cells with improved longevity, survival and/or persistence in vivo, such as for use in adoptive cell therapy methods.

[0352] 1. Chimeric Antigen Receptors (CARS)

[0353] In some embodiments, engineered cells, such as T cells, are provided that express a CAR with specificity for a particular antigen (or marker or ligand or receptor), such as an antigen expressed on the surface of a particular cell type.

[0354] In particular embodiments, the recombinant receptor, such as a chimeric receptor, contains an intracellular signaling region, which includes a cytoplasmic signaling domain (also interchangeably called an intracellular signaling domain), such as a cytoplasmic (intracellular) region capable of inducing a primary activation signal in a T cell, for example, a cytoplasmic signaling domain of a T cell receptor (TCR) component (e.g. a cytoplasmic signaling domain of a zeta chain of a CD3-zeta (CD3.zeta.) chain or a functional variant or signaling portion thereof) and/or that comprises an immunoreceptor tyrosine-based activation motif (ITAM).

[0355] In some embodiments, the chimeric receptor further contains an extracellular ligand-binding domain that specifically binds to a ligand (e.g. antigen) antigen. In some embodiments, the chimeric receptor is a CAR that contains an extracellular antigen-recognition domain that specifically binds to an antigen. In some embodiments, the ligand, such as an antigen, is a protein expressed on the surface of cells. The chimeric receptors, such as CARs, generally include an extracellular antigen binding domain, such as a portion of an antibody molecule, generally a variable heavy (V.sub.H) chain region and/or variable light (V.sub.L) chain region of the antibody, e.g., an scFv antibody fragment. In some embodiments, the CAR is a TCR-like CAR and the antigen is a processed peptide antigen, such as a peptide antigen of an intracellular protein, which, like a TCR, is recognized on the cell surface in the context of a major histocompatibility complex (MHC) molecule.

[0356] Exemplary antigen receptors, including CARs, and methods for engineering and introducing such receptors into cells, include those described, for example, in international patent application publication numbers WO200014257, WO2013126726, WO2012/129514, WO2014031687, WO2013/166321, WO2013/071154, WO2013/123061, WO2016/0046724, WO2016/014789, WO2016/090320, WO2016/094304, WO2017/025038, WO2017/173256, U.S. patent application publication numbers US2002131960, US2013287748, US20130149337, U.S. Pat. Nos. 6,451,995, 7,446,190, 8,252,592, 8,339,645, 8,398,282, 7,446,179, 6,410,319, 7,070,995, 7,265,209, 7,354,762, 7,446,191, 8,324,353, 8,479,118, and 9,765,342, and European patent application number EP2537416, and/or those described by Sadelain et al., Cancer Discov., 3(4): 388-398 (2013); Davila et al., PLoS ONE 8(4): e61338 (2013); Turtle et al., Curr. Opin. Immunol., 24(5): 633-39 (2012); Wu et al., Cancer, 18(2): 160-75 (2012). In some aspects, the antigen receptors include a CAR as described in U.S. Pat. No. 7,446,190, and those described in International Patent Application Publication No. WO/2014055668 A1. Examples of the CARs include CARs as disclosed in any of the aforementioned publications, such as WO2014031687, U.S. Pat. Nos. 8,339,645, 7,446,179, US 2013/0149337, U.S. Pat. Nos. 7,446,190, 8,389,282, Kochenderfer et al., Nature Reviews Clinical Oncology, 10, 267-276 (2013); Wang et al., J. Immunother. 35(9): 689-701 (2012); and Brentjens et al., Sci Transl Med. 5(177) (2013). See also WO2014031687, U.S. Pat. Nos. 8,339,645, 7,446,179, US 2013/0149337, U.S. Pat. Nos. 7,446,190, 8,389,282, WO 2016/090320, WO2016090327, WO2010104949A2 and WO2017173256.

[0357] In some embodiments, the CAR is constructed with a specificity for a particular antigen (or marker or ligand or receptor), such as an antigen expressed in a particular cell type to be targeted by adoptive therapy, e.g., a cancer marker, and/or an antigen intended to induce a dampening response, such as an antigen expressed on a normal or non-diseased cell type. Thus, the CAR typically includes in its extracellular portion one or more antigen binding molecules, such as one or more antigen-binding fragment, domain, or portion, or one or more antibody variable domains, and/or antibody molecules. In some embodiments, the CAR includes an antigen-binding portion or portions of an antibody molecule, such as a single-chain antibody fragment (scFv) derived from the variable heavy (VH) and variable light (VL) chains of a monoclonal antibody (mAb).

[0358] In some embodiments, the antibody or antigen-binding portion thereof is expressed on cells as part of a recombinant receptor, such as an antigen receptor. Among the antigen receptors are functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). Generally, a CAR containing an antibody or antigen-binding fragment that exhibits TCR-like specificity directed against peptide-MHC complexes also may be referred to as a TCR-like CAR. In some embodiments, the extracellular antigen binding domain specific for an MHC-peptide complex of a TCR-like CAR is linked to one or more intracellular signaling components, in some aspects via linkers and/or transmembrane domain(s). In some embodiments, such molecules can typically mimic or approximate a signal through a natural antigen receptor, such as a TCR, and, optionally, a signal through such a receptor in combination with a costimulatory receptor.

[0359] In some embodiments, the recombinant receptor, such as a chimeric receptor (e.g. CAR), includes a ligand-binding domain that binds, such as specifically binds, to an antigen (or a ligand or receptor). Among the antigens targeted by the chimeric receptors are those expressed in the context of a disease, condition, or cell type to be targeted via the adoptive cell therapy. Among the diseases and conditions are proliferative, neoplastic, and malignant diseases and disorders, including cancers and tumors, including hematologic cancers, cancers of the immune system, such as lymphomas, leukemias, and/or myelomas, such as B, T, and myeloid leukemias, lymphomas, and multiple myelomas.

[0360] In some embodiments, the antigen targeted by the receptor is a polypeptide. In some embodiments, it is a carbohydrate or other molecule. In some embodiments, the antigen is selectively expressed or overexpressed on cells of the disease or condition, e.g., the tumor or pathogenic cells, as compared to normal or non-targeted cells or tissues. In other embodiments, the antigen is expressed on normal cells and/or is expressed on the engineered cells.

[0361] In some embodiments, the CAR contains an antibody or an antigen-binding fragment (e.g. scFv) that specifically recognizes an antigen, such as an intact antigen, expressed on the surface of a cell.

[0362] In some embodiments, the antigen (or a ligand or receptor) is a tumor antigen or cancer marker. In certain embodiments, the antigen is or includes .alpha.v.beta.6 integrin (avb6 integrin), B cell maturation antigen (BCMA), B7-H3, B7-H6, carbonic anhydrase 9 (CA9, also known as CAIX or G250), a cancer-testis antigen, cancer/testis antigen 1B (CTAG, also known as NY-ESO-1 and LAGE-2), carcinoembryonic antigen (CEA), a cyclin, cyclin A2, C-C Motif Chemokine Ligand 1 (CCL-1), CD19, CD20, CD22, CD23, CD24, CD30, CD33, CD38, CD44, CD44v6, CD44v7/8, CD123, CD133, CD138, CD171, chondroitin sulfate proteoglycan 4 (CSPG4), epidermal growth factor protein (EGFR), type III epidermal growth factor receptor mutation (EGFR vIII), epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), ephrinB2, ephrine receptor A2 (EPHa2), estrogen receptor, Fc receptor like 5 (FCRL5; also known as Fc receptor homolog 5 or FCRH5), fetal acetylcholine receptor (fetal AchR), a folate binding protein (FBP), folate receptor alpha, ganglioside GD2, O-acetylated GD2 (OGD2), ganglioside GD3, glycoprotein 100 (gp100), glypican-3 (GPC3), G Protein Coupled Receptor 5D (GPCR5D), Her2/neu (receptor tyrosine kinase erb-B2), Her3 (erb-B3), Her4 (erb-B4), erbB dimers, Human high molecular weight-melanoma-associated antigen (HMW-MAA), hepatitis B surface antigen, Human leukocyte antigen A1 (HLA-A1), Human leukocyte antigen A2 (HLA-A2), IL-22 receptor alpha(IL-22Ra), IL-13 receptor alpha 2 (IL-13Ra2), kinase insert domain receptor (kdr), kappa light chain, L1 cell adhesion molecule (L1-CAM), CE7 epitope of L1-CAM, Leucine Rich Repeat Containing 8 Family Member A (LRRC8A), Lewis Y, Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, MAGE-A10, mesothelin (MSLN), c-Met, murine cytomegalovirus (CMV), mucin 1 (MUC1), MUC16, natural killer group 2 member D (NKG2D) ligands, melan A (MART-1), neural cell adhesion molecule (NCAM), oncofetal antigen, Preferentially expressed antigen of melanoma (PRAME), progesterone receptor, a prostate specific antigen, prostate stem cell antigen (PSCA), prostate specific membrane antigen (PSMA), Receptor Tyrosine Kinase Like Orphan Receptor 1 (ROR1), survivin, Trophoblast glycoprotein (TPBG also known as 5T4), tumor-associated glycoprotein 72 (TAG72), Tyrosinase related protein 1 (TRP1, also known as TYRP1 or gp75), Tyrosinase related protein 2 (TRP2, also known as dopachrome tautomerase, dopachrome delta-isomerase or DCT), vascular endothelial growth factor receptor (VEGFR), vascular endothelial growth factor receptor 2 (VEGFR2), Wilms Tumor 1 (WT-1), a pathogen-specific or pathogen-expressed antigen, or an antigen associated with a universal tag, and/or biotinylated molecules, and/or molecules expressed by HIV, HCV, HBV or other pathogens. Antigens targeted by the receptors in some embodiments include antigens associated with a B cell malignancy, such as any of a number of known B cell marker. In some embodiments, the antigen is or includes BCMA, CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igkappa, Iglambda, CD79a, CD79b or CD30.

[0363] In some embodiments, the antigen (or a ligand) is or includes orphan tyrosine kinase receptor ROR1, B cell maturation antigen (BCMA), carbonic anhydrase 9 (CAIX), Her2/neu (receptor tyrosine kinase erbB2), L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, epithelial glycoprotein 2 (EPG-2), epithelial glycoprotein 40 (EPG-40), EPHa2, erb-B2, erb-B3, erb-B4, erbB dimers, EGFR vIII, folate binding protein (FBP), FCRL5, FCRH5, fetal acetylcholine receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kinase insert domain receptor (kdr), kappa light chain, Lewis Y, L1-cell adhesion molecule, (L1-CAM), Melanoma-associated antigen (MAGE)-A1, MAGE-A3, MAGE-A6, Preferentially expressed antigen of melanoma (PRAME), survivin, TAG72, B7-H6, IL-13 receptor alpha 2 (IL-13Ra2), CA9, GD3, HMW-MAA, CD171, G250/CAIX, HLA-AI MAGE A1, HLA-A2 NY-ESO-1, PSCA, folate receptor-a, CD44v6, CD44v7/8, avb6 integrin, 8H9, NCAM, VEGF receptors, 5T4, Foetal AchR, NKG2D ligands, CD44v6, dual antigen, a cancer-testes antigen, mesothelin, murine CMV, mucin 1 (MUC1), MUC16, PSCA, NKG2D, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, O-acetylated GD2 (OGD2), CE7, Wilms Tumor 1 (WT-1), a cyclin, cyclin A2, CCL-1, CD138, a pathogen-specific antigen and an antigen associated with a universal tag, and/or biotinylated molecules, and/or molecules expressed by HIV, HCV, HBV or other pathogens. Antigens targeted by the receptors in some embodiments include antigens associated with a B cell malignancy, such as any of a number of known B cell marker. In some embodiments, the antigen targeted by the receptor is CD20, CD19, CD22, ROR1, CD45, CD21, CD5, CD33, Igkappa, Iglambda, CD79a, CD79b or CD30.

[0364] Antigens targeted by the receptors in some embodiments include orphan tyrosine kinase receptor ROR1, Her2, L1-CAM, CD19, CD20, CD22, mesothelin, CEA, and hepatitis B surface antigen, anti-folate receptor, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, 3, or 4, FBP, fetal acethycholine e receptor, GD2, GD3, HMW-MAA, IL-22R-alpha, IL-13R-alpha2, kdr, kappa light chain, Lewis Y, L1-cell adhesion molecule, MAGE-A1, mesothelin, MUC1, MUC16, PSCA, NKG2D Ligands, NY-ESO-1, MART-1, gp100, oncofetal antigen, ROR1, TAG72, VEGF-R2, carcinoembryonic antigen (CEA), prostate specific antigen, PSMA, Her2/neu, estrogen receptor, progesterone receptor, ephrinB2, CD123, c-Met, GD-2, and MAGE A3, CE7, Wilms Tumor 1 (WT-1), a cyclin, such as cyclin A1 (CCNA1), and/or biotinylated molecules, and/or molecules expressed by HIV, HCV, HBV or other pathogens.

[0365] In some embodiments, the CAR binds a pathogen-specific or pathogen-expressed antigen. In some embodiments, the CAR is specific for viral antigens (such as HIV, HCV, HBV, etc.), bacterial antigens, and/or parasitic antigens.

[0366] In some embodiments, the CAR contains a TCR-like antibody, such as an antibody or an antigen-binding fragment (e.g. scFv) that specifically recognizes an intracellular antigen, such as a tumor-associated antigen, presented on the cell surface as a MHC-peptide complex. In some embodiments, an antibody or antigen-binding portion thereof that recognizes an MHC-peptide complex can be expressed on cells as part of a recombinant receptor, such as an antigen receptor. Among the antigen receptors are functional non-TCR antigen receptors, such as chimeric antigen receptors (CARs). Generally, a CAR containing an antibody or antigen-binding fragment that exhibits TCR-like specificity directed against peptide-MHC complexes also may be referred to as a TCR-like CAR.

[0367] Reference to "Major histocompatibility complex" (MHC) refers to a protein, generally a glycoprotein, that contains a polymorphic peptide binding site or binding groove that can, in some cases, complex with peptide antigens of polypeptides, including peptide antigens processed by the cell machinery. In some cases, MHC molecules can be displayed or expressed on the cell surface, including as a complex with peptide, i.e. MHC-peptide complex, for presentation of an antigen in a conformation recognizable by an antigen receptor on T cells, such as a TCRs or TCR-like antibody. Generally, MHC class I molecules are heterodimers having a membrane spanning a chain, in some cases with three a domains, and a non-covalently associated .beta.2 microglobulin. Generally, MHC class II molecules are composed of two transmembrane glycoproteins, .alpha. and .beta., both of which typically span the membrane. An MHC molecule can include an effective portion of an MHC that contains an antigen binding site or sites for binding a peptide and the sequences necessary for recognition by the appropriate antigen receptor. In some embodiments, MHC class I molecules deliver peptides originating in the cytosol to the cell surface, where a MHC-peptide complex is recognized by T cells, such as generally CD8.sup.+ T cells, but in some cases CD4+ T cells. In some embodiments, MHC class II molecules deliver peptides originating in the vesicular system to the cell surface, where they are typically recognized by CD4.sup.+ T cells. Generally, MHC molecules are encoded by a group of linked loci, which are collectively termed H-2 in the mouse and human leukocyte antigen (HLA) in humans. Hence, typically human MHC can also be referred to as human leukocyte antigen (HLA).

[0368] The term "MHC-peptide complex" or "peptide-MHC complex" or variations thereof, refers to a complex or association of a peptide antigen and an MHC molecule, such as, generally, by non-covalent interactions of the peptide in the binding groove or cleft of the MHC molecule. In some embodiments, the MHC-peptide complex is present or displayed on the surface of cells. In some embodiments, the MHC-peptide complex can be specifically recognized by an antigen receptor, such as a TCR, TCR-like CAR or antigen-binding portions thereof.

[0369] In some embodiments, a peptide, such as a peptide antigen or epitope, of a polypeptide can associate with an MHC molecule, such as for recognition by an antigen receptor. Generally, the peptide is derived from or based on a fragment of a longer biological molecule, such as a polypeptide or protein. In some embodiments, the peptide typically is about 8 to about 24 amino acids in length. In some embodiments, a peptide has a length of from or from about 9 to 22 amino acids for recognition in the MHC Class II complex. In some embodiments, a peptide has a length of from or from about 8 to 13 amino acids for recognition in the MHC Class I complex. In some embodiments, upon recognition of the peptide in the context of an MHC molecule, such as MHC-peptide complex, the antigen receptor, such as TCR or TCR-like CAR, produces or triggers an activation signal to the T cell that induces a T cell response, such as T cell proliferation, cytokine production, a cytotoxic T cell response or other response.

[0370] In some embodiments, a TCR-like antibody or antigen-binding portion, are known or can be produced by methods known in the art (see e.g. US Published Application Nos. US 2002/0150914; US 2003/0223994; US 2004/0191260; US 2006/0034850; US 2007/00992530; US20090226474; US20090304679; and International PCT Publication No. WO 03/068201).

[0371] In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to a MHC-peptide complex, can be produced by immunizing a host with an effective amount of an immunogen containing a specific MHC-peptide complex. In some cases, the peptide of the MHC-peptide complex is an epitope of antigen capable of binding to the MHC, such as a tumor antigen, for example a universal tumor antigen, myeloma antigen or other antigen. In some embodiments, an effective amount of the immunogen is then administered to a host for eliciting an immune response, wherein the immunogen retains a three-dimensional form thereof for a period of time sufficient to elicit an immune response against the three-dimensional presentation of the peptide in the binding groove of the MHC molecule. Serum collected from the host is then assayed to determine if desired antibodies that recognize a three-dimensional presentation of the peptide in the binding groove of the MHC molecule is being produced. In some embodiments, the produced antibodies can be assessed to confirm that the antibody can differentiate the MHC-peptide complex from the MHC molecule alone, the peptide of interest alone, and a complex of MHC and irrelevant peptide. The desired antibodies can then be isolated.

[0372] In some embodiments, an antibody or antigen-binding portion thereof that specifically binds to an MHC-peptide complex can be produced by employing antibody library display methods, such as phage antibody libraries. In some embodiments, phage display libraries of mutant Fab, scFv or other antibody forms can be generated, for example, in which members of the library are mutated at one or more residues of a CDR or CDRs. See e.g. US published application No. US20020150914, US2014/0294841; and Cohen C J. et al. (2003) J Mol. Recogn. 16:324-332.

[0373] The term "antibody" herein is used in the broadest sense and includes polyclonal and monoclonal antibodies, including intact antibodies and functional (antigen-binding) antibody fragments, including fragment antigen binding (Fab) fragments, F(ab').sub.2 fragments, Fab' fragments, Fv fragments, recombinant IgG (rIgG) fragments, variable heavy chain (V.sub.H) regions capable of specifically binding the antigen, single chain antibody fragments, including single chain variable fragments (scFv), and single domain antibodies (e.g., sdAb, sdFv, nanobody) fragments. The term encompasses genetically engineered and/or otherwise modified forms of immunoglobulins, such as intrabodies, peptibodies, chimeric antibodies, fully human antibodies, humanized antibodies, and heteroconjugate antibodies, multispecific, e.g., bispecific, antibodies, diabodies, triabodies, and tetrabodies, tandem di-scFv, tandem tri-scFv. Unless otherwise stated, the term "antibody" should be understood to encompass functional antibody fragments thereof. The term also encompasses intact or full-length antibodies, including antibodies of any class or sub-class, including IgG and sub-classes thereof, IgM, IgE, IgA, and IgD.

[0374] In certain embodiments, multispecific binding molecules, e.g., multispecific chimeric receptors, such as multispecific CARs, can contain any of the multispecific antibodies, including, e.g. bispecific antibodies, multispecific single-chain antibodies, e.g., diabodies, triabodies, and tetrabodies, tandem di-scFvs, and tandem tri-scFvs.

[0375] In some embodiments, the antigen-binding proteins, antibodies and antigen binding fragments thereof specifically recognize an antigen of a full-length antibody. In some embodiments, the heavy and light chains of an antibody can be full-length or can be an antigen-binding portion (a Fab, F(ab')2, Fv or a single chain Fv fragment (scFv)). In other embodiments, the antibody heavy chain constant region is chosen from, e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE, particularly chosen from, e.g., IgG1, IgG2, IgG3, and IgG4, more particularly, IgG1 (e.g., human IgG1). In another embodiment, the antibody light chain constant region is chosen from, e.g., kappa or lambda, particularly kappa.

[0376] Among the provided antibodies are antibody fragments. An "antibody fragment" or "antigen-binding fragment" refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds. Examples of antibody fragments include but are not limited to Fv, Fab, Fab', Fab'-SH, F(ab').sub.2; diabodies; linear antibodies; variable heavy chain (V.sub.H) regions, single-chain antibody molecules such as scFvs and single-domain V.sub.H single antibodies; and multispecific antibodies formed from antibody fragments. In some embodiments, the antigen-binding domain in the provided CARs is or comprises an antibody fragment comprising a variable heavy chain (V.sub.H) and a variable light chain (V.sub.L) region. In particular embodiments, the antibodies are single-chain antibody fragments comprising a variable heavy chain region and/or a variable light chain region, such as scFvs.

[0377] The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to antigen. The variable domains of the heavy chain and light chain (V.sub.H and V.sub.L, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved framework regions (FRs) and three CDRs. (See, e.g., Kindt et al. Kuby Immunology, 6th ed., W.H. Freeman and Co., page 91 (2007). A single V.sub.H or V.sub.L domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind a particular antigen may be isolated using a V.sub.H or V.sub.L domain from an antibody that binds the antigen to screen a library of complementary V.sub.L or V.sub.H domains, respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991).

[0378] The terms "complementarity determining region," and "CDR," synonymous with "hypervariable region" or "HVR," are known in the art to refer to non-contiguous sequences of amino acids within antibody variable regions, which confer antigen specificity and/or binding affinity. In general, there are three CDRs in each heavy chain variable region (CDR-H1, CDR-H2, CDR-H3) and three CDRs in each light chain variable region (CDR-L1, CDR-L2, CDR-L3). "Framework regions" and "FR" are known in the art to refer to the non-CDR portions of the variable regions of the heavy and light chains. In general, there are four FRs in each full-length heavy chain variable region (FR-H1, FR-H2, FR-H3, and FR-H4), and four FRs in each full-length light chain variable region (FR-L1, FR-L2, FR-L3, and FR-L4).

[0379] The precise amino acid sequence boundaries of a given CDR or FR can be readily determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. ("Kabat" numbering scheme); Al-Lazikani et al., (1997) JMB 273,927-948 ("Chothia" numbering scheme); MacCallum et al., J. Mol. Biol. 262:732-745 (1996), "Antibody-antigen interactions: Contact analysis and binding site topography," J. Mol. Biol. 262, 732-745." ("Contact" numbering scheme); Lefranc M P et al., "IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains," Dev Comp Immunol, 2003 January; 27(1):55-77 ("IMGT" numbering scheme); Honegger A and Pluckthun A, "Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool," J Mol Biol, 2001 Jun. 8; 309(3):657-70, ("Aho" numbering scheme); and Martin et al., "Modeling antibody hypervariable loops: a combined algorithm," PNAS, 1989, 86(23):9268-9272, ("AbM" numbering scheme).

[0380] The boundaries of a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignments, while the Chothia scheme is based on structural information. Numbering for both the Kabat and Chothia schemes is based upon the most common antibody region sequence lengths, with insertions accommodated by insertion letters, for example, "30a," and deletions appearing in some antibodies. The two schemes place certain insertions and deletions ("indels") at different positions, resulting in differential numbering. The Contact scheme is based on analysis of complex crystal structures and is similar in many respects to the Chothia numbering scheme. The AbM scheme is a compromise between Kabat and Chothia definitions based on that used by Oxford Molecular's AbM antibody modeling software.

[0381] Table 1, below, lists exemplary position boundaries of CDR-L1, CDR-L2, CDR-L3 and CDR-H1, CDR-H2, CDR-H3 as identified by Kabat, Chothia, AbM, and Contact schemes, respectively. For CDR-H1, residue numbering is listed using both the Kabat and Chothia numbering schemes. FRs are located between CDRs, for example, with FR-L1 located before CDR-L1, FR-L2 located between CDR-L1 and CDR-L2, FR-L3 located between CDR-L2 and CDR-L3 and so forth. It is noted that because the shown Kabat numbering scheme places insertions at H35A and H35B, the end of the Chothia CDR-H1 loop when numbered using the shown Kabat numbering convention varies between H32 and H34, depending on the length of the loon.


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