TREATMENT OF CANCER USING HUMANIZED ANTI-BCMA CHIMERIC ANTIGEN RECEPTOR

• US 20160046724A1
• Filed: 07/21/2015
• Published: 02/18/2016
• Est. Priority Date: 07/21/2014
• Status: Active Grant

• Current Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Novartis Ag, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Brogdon, Jennifer, Glass, David, June, Carl H., Mannick, Joan, Milone, Michael C., Murphy, Leon, Ruella, Marco, Choi, Eugene, Ebersbach, Hilmar Erhard, Huet, Heather, Plesa, Gabriela, Richardson, Celeste, Singh, Reshma, Wang, Yongqiang, Wu, Qilong
• Granted Patent: US 10,174,095 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 38/177 Receptors; Cell surface antigens; Cell surface determinants
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70578 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 14/7151 for tumor necrosis factor [TNF], for lymphotoxin [LT]
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: C07K 2319/74 : containing a fusion for binding to a cell surface receptor
• CPC Class Codes: C12N 15/63 : Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 5/00 : Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor; plant reproduction by tissue culture techniques A01H4/00

Abstract

The invention provides compositions and methods for treating diseases associated with expression of BCMA. The invention also relates to chimeric antigen receptor (CAR) specific to BCMA vectors encoding the same, and recombinant T cells comprising the BCMA CAR. The invention also includes methods of administering a genetically modified T cell expressing a CAR that comprises a BCMA binding domain.

Claims

1. An isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an anti-BCMA binding domain, a transmembrane domain, and an intracellular signaling domain, and wherein said anti-BCMA binding domain comprises:

(i) a heavy chain complementary determining region 1 (HC CDR1), a heavy chain complementary determining region 2 (HC CDR2), and a heavy chain complementary determining region 3 (HC CDR3) of any anti-BMCA heavy chain binding domain amino acid sequences listed in Table 1 or 16; and

(ii) a light chain complementary determining region 1 (LC CDR1), a light chain complementary determining region 2 (LC CDR2), and a light chain complementary determining region 3 (LC CDR3) of any anti-BMCA light chain binding domain amino acid sequences listed in Table 1 or 16.

3. The isolated nucleic acid molecule of claim 1, wherein said LC CDR1, LC CDR2, and LC CDR3 are the LC CDR sequences listed in Table 21, 23, or 25.

4. The isolated nucleic acid molecule of claim 1, wherein said HC CDR1, HC CDR2, and HC CDR3 are the HC CDR sequences listed in Table 20, 22, or 24.

5. The isolated nucleic acid molecule of claim 1, which encodes a CAR comprising:

(i) the amino acid sequence of any light chain variable region listed in Table 1;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of the amino acid sequence of any of the light chain variable regions provided in Table 1;

or

(iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any of the light chain variable regions provided in Table 1.

6. The isolated nucleic acid molecule of claim 1, which encodes a CAR comprising:

(i) the amino acid sequence of any heavy chain variable region listed in Table 1;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of the amino acid sequence of any of the heavy chain variable regions provided in Table 1;

or

(iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any of the heavy chain variable regions provided in Table 1.

7. The isolated nucleic acid molecule of claim 1, which encodes a CAR comprising the amino acid sequence of any light chain variable region listed in Table 1, and the amino acid sequence of any heavy chain variable region listed in Table 1.

8. The isolated nucleic acid molecule of claim 1, wherein the encoded anti-BCMA binding domain comprises:

(i) the amino acid sequence selected from a group consisting of SEQ ID NO;

49, 39-53, 69-83, 84-98, 129-149, 171-191, 192-212, 255-258, 259-262, 263-266, 271, and 273;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any of SEQ ID NO;

49, 39-53, 69-83, 84-98, 129-149, 171-191, 192-212, 255-258, 259-262, 263-266, 271, or 273;

or

(iii) an amino acid sequence with 95-99% identity to any of SEQ ID NO;

49, 39-53, 69-83, 84-98, 129-149, 171-191, 192-212, 255-258, 259-262, 263-266, 271, or 273.

9. The isolated nucleic acid molecule of claim 1, wherein the anti-BCMA binding domain comprises a nucleotide sequence selected from a group consisting of SEQ ID NO:

64, 54-68, 150-170, 272, and 274, or a sequence with 95-99% identity thereof.

10. The isolated nucleic acid molecule of claim 1, wherein:

(i) the encoded CAR comprises a transmembrane domain that comprises a transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CDS, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154;

or

(ii) the encoded transmembrane domain comprises the amino acid sequence of SEQ ID NO;

6, an amino acid sequence comprising at least one, two or three modifications but not more than 20, 10 or 5 modifications of the amino acid sequence of SEQ ID NO;

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

6;

or (ii) the nucleic acid sequence encoding the transmembrane domain comprises the nucleotide sequence of SEQ ID NO;

17, or the nucleotide sequence with 95-99% identity thereof.

11. The isolated nucleic acid molecule of claim 1, wherein the encoded anti-BCMA binding domain comprises:

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

49;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to SEQ ID NO;

49;

or

(iii) an amino acid sequence with 95-99% identity to SEQ ID NO;

49.

12. The isolated nucleic acid molecule of claim 1, wherein the encoded anti-BCMA binding domain is connected to the transmembrane domain by a hinge region, wherein (i) the encoded hinge region comprises the amino acid sequence of SEQ ID NO:

2, or a sequence with 95-99% identity thereof;

or (ii) the nucleic acid sequence encoding the hinge region comprises the nucleotide sequence of SEQ ID NO;

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

14. The isolated nucleic acid molecule of claim 1, wherein the encoded intracellular domain comprises a costimulatory domain, wherein the costimulatory domain comprises a functional signaling domain derived from a protein selected from the group consisting of MHC class I molecules, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds with CD83.

15. The isolated nucleic acid molecule of claim 14, wherein the encoded costimulatory domain comprises the amino acid sequence of SEQ ID NO:

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

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

7.

16. The isolated nucleic acid molecule of claim 14, wherein the nucleic acid sequence encoding the costimulatory domain comprises the nucleotide sequence of SEQ ID NO:

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

17. The isolated nucleic acid molecule of claim 1, wherein the encoded intracellular signaling domain comprises a functional signaling domain of 4-1BB and a functional signaling domain of CD3 zeta.

18. The isolated nucleic acid molecule of claim 1, wherein the encoded intracellular signaling domain comprises the amino acid sequence of SEQ ID NO:

7 and the sequence of SEQ ID NO;

9 or SEQ ID NO;

10;

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

7 and the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

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

7 and the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

10.

19. The isolated nucleic acid molecule of claim 1, wherein the encoded intracellular signaling domain comprises the sequence of SEQ ID NO:

7 and the sequence of SEQ ID NO;

9 or SEQ ID NO;

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

20. The isolated nucleic acid molecule of claim 1, wherein the nucleic acid sequence encoding the intracellular signaling domain comprises the sequence of SEQ ID NO:

18, or a sequence with 95-99% identity thereof, and/or the sequence of SEQ ID NO;

20 or SEQ ID NO;

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

21. The isolated nucleic acid molecule of claim 1, further comprising a leader sequence which encodes the amino acid sequence of SEQ ID NO:

1.

22. The isolated nucleic acid molecule of claim 1, which encodes a CAR comprising:

(i) the amino acid sequence of any of SEQ ID NOs;

109, 99-113, 213-233, or 267-270;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any of SEQ ID NOs;

109, 99-113, 213-233, or 267-270;

or

(iii) an amino acid sequence with 95-99% identity to any of SEQ ID NOs;

109, 99-113, 213-233, or 267-270.

23. The isolated nucleic acid molecule of claim 1, comprising the nucleotide sequence of any of SEQ ID NOs:

124, 114-128 or 234-254, or a nucleotide sequence with 95-99% identity to any of SEQ ID NOs;

124, 114-128 or 234-254.

44. A vector comprising a nucleic acid molecule encoding a CAR polypeptide of claim 1, wherein the vector is a DNA vector, a RNA vector, a plasmid, a lentivirus vector, adenoviral vector, or a retrovirus vector.

46. An immune effector cell comprising the nucleic acid molecule of claim 1.

47. A method of making an immune effector cell, comprising transducing the immune effector cell with the vector of claim 44.

48. A method of generating a population of RNA-engineered cells comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, where the RNA comprises the nucleic acid molecule of claim 1.

49. A method of providing an anti-tumor immunity in a mammal comprising administering to the mammal an effective amount of a cell comprising the CAR nucleic acid molecule of claim 1.

51. A method of treating a mammal having a disease associated with expression of BCMA, comprising administering to the mammal an effective amount of a cell comprising the CAR nucleic acid molecule of claim 1.

52. The method of claim 51, wherein the disease associated with BCMA expression is:

(i) a cancer or malignancy, or a precancerous condition chosen from one or more of a myelodysplasia, a myelodysplastic syndrome or a preleukemia, or

(ii) a non-cancer related indication associated with expression of BCMA.

54. The method of claim 51, wherein the disease is an acute leukemia chosen from one or more of B-cell acute lymphoid leukemia (“

BALL”

), T-cell acute lymphoid leukemia (“

TALL”

), acute lymphoid leukemia (ALL);

chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL);

B cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt'"'"'s lymphoma, diffuse large B cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell- or a large cell-follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, Marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin'"'"'s lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia;

a prostate cancer, pancreatic cancer, lung cancer;

or a plasma cell proliferative disorder, monoclonal gammapathy of undetermined significance (MGUS), Waldenstrom'"'"'s macroglobulinemia, a plasmacytoma, systemic amyloid light chain amyloidosis, and POEMS syndrome, or a combination thereof.

55. The method of claim 51, wherein the cell is administered in combination with one or more of:

(i) an agent that increases the efficacy of the cell comprising the CAR nucleic acid or CAR polypeptide;

(ii) an agent that ameliorates one or more side effects associated with administration of the cell comprising the CAR nucleic acid or CAR polypeptide;

or

(iii) an agent that treats the disease associated with BCMA.

56. The method of claim 51, wherein the cell is administered in combination with a checkpoint inhibitor.

59. The cell of claim 46, further expressing a molecule that comprises a first polypeptide that comprises at least a portion of an inhibitory molecule, associated with a second polypeptide that comprises a positive signal from an intracellular signaling domain.

61. The method of claim 55, wherein the agent is an mTOR inhibitor and the subject is administered a low, immune enhancing, dose of an mTOR inhibitor.

63. The method of claim 51, wherein the cell is administered in combination with a cell comprising a CD19 CAR molecule.

64. The method of claim 63, wherein the disease associated with BCMA is multiple myeloma.

66. The method of claim 51, wherein the cell is administered to the subject by one, two, three, or more separate administrations of a partial dose,

optionally wherein a first percentage of the total dose is administered on a first day of treatment, a second percentage of the total dose is administered on a subsequent (e.g., second, third, fourth, fifth, sixth, or seventh or later) day of treatment, and optionally, a third percentage (e.g., the remaining percentage) of the total dose is administered on a yet subsequent (e.g., third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, or later) day of treatment,

optionally wherein 10% of the total dose of cells is administered on the first day, 30% of the total dose of cells is administered on the second day, and the remaining 60% of the total dose of cells is administered on the third day of treatment, and

optionally wherein the total cell dose comprises 1 to 5×

10⁷ or 1 to 5×

10⁸ cells.

67. The isolated nucleic acid molecule of claim 1, wherein the encoded intracellular signaling domain comprises a primary signaling domain comprising a functional signaling domain derived from CD3 zeta, TCR zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CDS, CD22, CD79a, CD79b, CD278 (also known as “

ICOS”

), Fcε

RI, DAP10, DAP12, or CD66d.

68. The isolated nucleic acid molecule of claim 67, wherein the encoded primary signaling domain comprises a functional signaling domain of CD3 zeta, wherein the CD3 zeta comprises the amino acid sequence of SEQ ID NO:

9 or SEQ ID NO;

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

9 or SEQ ID NO;

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

9 or SEQ ID NO;

10.

71. The isolated nucleic acid molecule of claim 1, wherein the encoded CAR is expressed in frame and as a single polypeptide chain.

2. (canceled)

13. (canceled)

24. (canceled)

25. An isolated chimeric antigen receptor (CAR) polypeptide, wherein the CAR comprises an anti-BCMA binding domain, a transmembrane domain, and an intracellular signaling domain, and wherein said anti-BCMA binding domain comprises:

(i) a heavy chain complementary determining region 1 (HC CDR1), a heavy chain complementary determining region 2 (HC CDR2), and a heavy chain complementary determining region 3 (HC CDR3) of any anti-BCMA heavy chain binding domain amino acid sequences listed in Table 1 or 16; and

(ii) a light chain complementary determining region 1 (LC CDR1), a light chain complementary determining region 2 (LC CDR2), and a light chain complementary determining region 3 (LC CDR3) of any anti-BCMA light chain binding domain amino acid sequences listed in Table 1 or 16.

27. The isolated CAR polypeptide of claim 25, wherein:

(i) said LC CDR1, LC CDR2, and LC CDR3 are the LC CDR sequences listed in Table 21, 23, or 25;

(ii) said HC CDR1, HC CDR2 and HC CDR3 are the HC CDR sequences listed in Table 21, 23, or 25;

or

(iii) both (i) and (ii).

29. The isolated CAR polypeptide of claim 25, comprising:

(i) the amino acid sequence of any light chain variable region listed in Table 1;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of the amino acid sequence of any of the light chain variable region provided in Table 1;

(iii) an amino acid sequence with 95-99% identity to the amino acid sequence of any of the light chain variable region provided in Table 1;

(iv) the amino acid sequence of any heavy chain variable region listed in Table 1;

(v) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications of the amino acid sequence of any of the heavy chain variable regions provided in Table 1;

or

(vi) an amino acid sequence with 95-99% identity to the amino acid sequence of any of the heavy chain variable regions provided in Table 1;

or

(vii) any of (i)-(iii) and any of (iv)-(vi).

31. The isolated CAR polypeptide of claim 25, comprising:

(i) the amino acid sequence of any light chain variable region listed in Table 1, and the amino acid sequence of any heavy chain variable region provided in Table 1;

(ii) an amino acid sequence selected from a group consisting of SEQ ID NO;

49, 39-53, 69-83, 84-98, 129-149, 171-191, 192-212, 255-258, 259-262, 263-266, 271, and 273;

(iii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any of SEQ ID NO;

49, 39-53, 69-83, 84-98, 129-149, 171-191, 192-212, 255-258, 259-262, 263-266, 271, and 273;

or

(iv) an amino acid sequence with 95-99% identity to any of SEQ ID NO;

49, 39-53, 69-83, 84-98, 129-149, 171-191, 192-212, 255-258, 259-262, 263-266, 271, and 273.

33. The isolated CAR polypeptide of claim 25, wherein the transmembrane domain comprises a transmembrane domain from a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CDS, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154.

34. The isolated CAR polypeptide of claim 25, wherein

(i) the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

6;

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

6;

or

(iii) a sequence with 95-99% identity to the amino acid sequence of SEQ ID NO;

6.

35. The isolated CAR polypeptide of claim 25, wherein the anti-BCMA binding domain is connected to the transmembrane domain by a hinge region, wherein the hinge region comprises SEQ ID NO:

2 or SEQ ID NO;

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

37. The isolated CAR polypeptide of claim 25, wherein the encoded intracellular domain comprises a costimulatory domain, wherein the costimulatory domain comprises a functional signaling domain derived from a protein selected from the group consisting of MHC class I molecules, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation molecules (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds with CD83.

38. The isolated CAR polypeptide of claim 25, wherein the costimulatory domain comprises the amino acid sequence of SEQ ID NO:

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

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

7.

39. The isolated CAR polypeptide of claim 25, wherein the intracellular signaling domain comprises a functional signaling domain of 4-1BB and a functional signaling domain of CD3 zeta.

40. The isolated CAR polypeptide of claim 25, wherein the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO:

7 and the sequence of SEQ ID NO;

9 or SEQ ID NO;

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

7 and the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

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

7 and the amino acid sequence of SEQ ID NO;

9 or SEQ ID NO;

10.

43. The isolated CAR polypeptide of claim 25, comprising:

(i) the amino acid sequence of any of SEQ ID NO;

109, 99-113, 213-233, or 267-270;

(ii) an amino acid sequence having at least one, two or three modifications but not more than 30, 20 or 10 modifications to any of SEQ ID NOs;

109, 99-113, 213-233, or 267-270;

or

(iii) an amino acid sequence with 95-99% identity to any of SEQ ID NO;

109, 99-113, 213-233, or 267-270.

69. The isolated CAR polypeptide of claim 25, wherein the intracellular signaling domain comprises a primary signaling domain comprising a functional signaling domain derived from CD3 zeta, TCR zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CDS, CD22, CD79a, CD79b, CD278 (also known as “

ICOS”

), Fcε

RI, DAP10, DAP12, or CD66d.

70. The isolated CAR polypeptide of claim 69, wherein the primary signaling domain comprises a functional signaling domain of CD3 zeta, wherein the CD3 zeta comprises the amino acid sequence of SEQ ID NO:

9 or SEQ ID NO;

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

9 or SEQ ID NO;

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

9 or SEQ ID NO;

10.

26. (canceled)

28. (canceled)

30. (canceled)

32. (canceled)

36. (canceled)

41. (canceled)

42. (canceled)

45. (canceled)

50. (canceled)

53. (canceled)

57. (canceled)

58. (canceled)

60. (canceled)

62. (canceled)

65. (canceled)