BCMA CHIMERIC ANTIGEN RECEPTORS

1 . A chimeric antigen receptor (CAR) comprising: an extracellular domain that comprises a humanized anti-BCMA (B cell maturation antigen) antibody or antigen binding fragment thereof that binds one or more epitopes of a human BCMA polypeptide; a transmembrane domain, one or more intracellular co-stimulatory signaling domains, and a primary signaling domain.

2 . The CAR of claim 1 , wherein the humanized anti-BCMA antibody or antigen binding fragment that binds the human BCMA polypeptide is selected from the group consisting of: a Camel Ig, Ig NAR, Fab fragments, Fab′ fragments, F(ab)′2 fragments, F(ab)′3 fragments, Fv, single chain Fv antibody (“scFv”), bis-scFv, (scFv)2, minibody, diabody, triabody, tetrabody, disulfide stabilized Fv protein (“dsFv”), and single-domain antibody (sdAb, Nanobody).

3 . The CAR of claim 2 , wherein the humanized anti-BCMA antibody or antigen binding fragment that binds the human BCMA polypeptide is an scFv.

4 . The CAR of any one of claims 1 to 3 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises one or more CDRs as set forth in any one of SEQ ID NOs: 1-3.

5 . The CAR of any one of claims 1 to 4 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises one or more CDRs as set forth in any one of SEQ ID NOs: 4-6.

6 . The CAR of any one of claims 1 to 5 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises a variable light chain sequence as set forth in any one of SEQ ID NOs: 7-9.

7 . The CAR of claim 6 , wherein the variable light chain sequence comprises the CDR sequences set forth in SEQ ID NOs: 1-3.

8 . The CAR of any one of claims 1 to 7 , wherein the humanized anti-BCMA antibody or antigen binding fragment thereof comprises a variable heavy chain sequence as set forth in any one of SEQ ID NOs: 10-14.

9 . The CAR of claim 8 , wherein the variable heavy chain sequence comprises the CDR sequences set forth in SEQ ID NOs: 4-6

10 . The CAR of any one of claims 1 to 9 , wherein the transmembrane domain is from a polypeptide selected from the group consisting of: alpha, beta or zeta chain of the T-cell receptor, CD3ε, CD3ζ, CD4, CD5, CD8α, CD9, CD 16, CD22, CD27, CD28, CD33, CD37, CD45, CD64, CD80, CD86, CD 134, CD137, CD152, CD 154, and PD1.

11 . The CAR of any one of claims 1 to 9 , wherein the transmembrane domain is from a polypeptide selected from the group consisting of: CD8α; CD4, CD45, PD1, and CD152.

12 . The CAR of any one of claims 1 to 11 , wherein the transmembrane domain is from CD8α.

13 . The CAR of any one of claims 1 to 11 , wherein the transmembrane domain is from PD1.

14 . The CAR of any one of claims 1 to 11 , wherein the transmembrane domain is from CD152.

15 . The CAR of any one of claims 1 to 14 , wherein the one or more co-stimulatory signaling domains are from a co-stimulatory molecule selected from the group consisting of: CARD11, CD2, CD7, CD27, CD28, CD30, CD40, CD54 (ICAM), CD83, CD134 (OX40), CD137 (4-1BB), CD150 (SLAMF1), CD152 (CTLA4), CD223 (LAG3), CD270 (HVEM), CD273 (PD-L2), CD274 (PD-L1), CD278 (ICOS), DAP10, LAT, NKD2C SLP76, TRIM, and ZAP70.

16 . The CAR of any one of claims 1 to 14 , wherein the one or more co-stimulatory signaling domains are from a co-stimulatory molecule selected from the group consisting of: CD28, CD134, and CD137.

17 . The CAR of any one of claims 1 to 14 , wherein the one or more co-stimulatory signaling domains are from a co-stimulatory molecule selected from the group consisting of: CD28, CD134, and CD137.

18 . The CAR of any one of claims 1 to 14 , wherein the one or more co-stimulatory signaling domains is from CD28.

19 . The CAR of any one of claims 1 to 14 , wherein the one or more co-stimulatory signaling domains is from CD134.

20 . The CAR of any one of claims 1 to 14 , wherein the one or more co-stimulatory signaling domains is from CD137.

21 . The CAR of any one of claims 1 to 20 , further comprising a hinge region polypeptide.

22 . The CAR of claim 21 , wherein the hinge region polypeptide comprises a hinge region of CD8α.

23 . The CAR of claim 21 , wherein the hinge region polypeptide comprises a hinge region of PD1.

24 . The CAR of claim 21 , wherein the hinge region polypeptide comprises a hinge region of CD152.

25 . The CAR of any one of claims 1 to 24 , further comprising a spacer region.

26 . The CAR of claim 25 , wherein the spacer region polypeptide comprises CH2 and CH3 regions of IgG1 or IgG4.

27 . The CAR of any one of claims 1 to 26 , further comprising a signal peptide.

28 . The CAR of claim 27 , wherein the signal peptide comprises an IgG1 heavy chain signal polypeptide, a CD8α signal polypeptide, or a human GM-CSF receptor alpha signal polypeptide.

29 . The CAR of any one of claims 1 to 28 , comprising an amino acid sequence as set forth in any one of SEQ ID NOs: 15-29, 71, and 73.

30 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 15.

31 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 16.

32 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 17.

33 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 18.

34 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 19.

35 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 20.

36 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 21.

37 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 22.

38 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 23.

39 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 24.

40 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 25.

41 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 26.

42 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 27.

43 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 28.

44 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 29.

45 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 71.

46 . The CAR of any one of claims 1 to 29 , comprising an amino acid sequence as set forth in SEQ ID NO: 73.

47 . A polynucleotide encoding a CAR of any one of claims 1 to 46 .

48 . A polynucleotide encoding a CAR, wherein the polynucleotide sequence is set forth in any one of SEQ ID NOs: 30-44, 70, and 72.

49 . A vector comprising the polynucleotide of claim 47 or claim 48 .

50 . The vector of claim 49 , wherein the vector is an expression vector.

51 . The vector of claim 49 , wherein the vector is an episomal vector.

52 . The vector of claim 49 , wherein the vector is a viral vector.

53 . The vector of claim 49 , wherein the vector is a retroviral vector.

54 . The vector of claim 49 , wherein the vector is a lentiviral vector.

55 . The vector of claim 49 , wherein the lentiviral vector is selected from the group consisting essentially of: human immunodeficiency virus 1 (HIV-1); human immunodeficiency virus 2 (HIV-2), visna-maedi virus (VMV) virus; caprine arthritis-encephalitis virus (CAEV); equine infectious anemia virus (EIAV); feline immunodeficiency virus (FIV); bovine immune deficiency virus (BIV); and simian immunodeficiency virus (SIV).

56 . The vector according to any one of claims 52 to 55 , comprising a left (5′) retroviral LTR, a Psi (Ψ) packaging signal, a central polypurine tract/DNA flap (cPPT/FLAP), a retroviral export element; a promoter operably linked to the polynucleotide of claim 26 or claim 27 ; and a right (3′) retroviral LTR.

57 . The vector of claim 56 , further comprising a heterologous polyadenylation sequence.

58 . The vector of claim 56 or claim 57 , further comprising a hepatitis B virus posttranscriptional regulatory element (HPRE) or woodchuck post-transcriptional regulatory element (WPRE).

59 . The vector of any one of claims 56 to 58 , wherein the promoter of the 5′ LTR is replaced with a heterologous promoter.

60 . The vector of claim 59 , wherein the heterologous promoter is a cytomegalovirus (CMV) promoter, a Rous Sarcoma Virus (RSV) promoter, or an Simian Virus 40 (SV40) promoter.

61 . The vector of any one of claims 56 to 60 , wherein the 5′ LTR or 3′ LTR is a lentivirus LTR.

62 . The vector of any one of claims 56 to 61 , wherein the 3′ LTR comprises one or more modifications.

63 . The vector of any one of claims 56 to 62 , wherein the 3′ LTR comprises one or more deletions.

64 . The vector of any one of claims 56 to 63 , wherein the 3′ LTR is a self-inactivating (SIN) LTR.

65 . The vector of any one of claims 56 to 64 , wherein the polyadenylation sequence is a bovine growth hormone polyadenylation or signal rabbit β-globin polyadenylation sequence.

66 . The vector of any one of claims 59 to 65 , wherein the polynucleotide of claim 57 or claim 58 comprises an optimized Kozak sequence.

67 . The vector of any one of claims 56 to 66 , wherein the promoter operably linked to the polynucleotide of claim 57 or claim 58 is selected from the group consisting of: a cytomegalovirus immediate early gene promoter (CMV), an elongation factor 1 alpha promoter (EF1-α), a phosphoglycerate kinase-1 promoter (PGK), a ubiquitin-C promoter (UBQ-C), a cytomegalovirus enhancer/chicken beta-actin promoter (CAG), polyoma enhancer/herpes simplex thymidine kinase promoter (MC1), a beta actin promoter (β-ACT), a simian virus 40 promoter (SV40), and a myeloproliferative sarcoma virus enhancer, negative control region deleted, d1587rev primer-binding site substituted (MND) promoter.

68 . An immune effector cell comprising the vector of any one of claims 49 to 67 .

69 . The immune effector cell of claim 68 , wherein the immune effector cell is selected from the group consisting of: a T lymphocyte and a natural killer (NK) cell.

70 . A composition comprising the immune effector cell of claim 68 or claim 69 and a physiologically acceptable excipient.

71 . A method of generating an immune effector cell comprising a CAR according to any one of claims 1 to 46 comprising introducing into an immune effector cell the vector of an one of claims 49 to 67 .

72 . The method of claim 71 , further comprising stimulating the immune effector cell and inducing the cell to proliferate by contacting the cell with antibodies that bind CD3 and antibodies that bind to CD28; thereby generating a population of immune effector cells.

73 . The method of claim 72 , wherein the immune effector cell is stimulated and induced to proliferate before introducing the vector.

74 . The method of claim 72 , wherein the immune effector cells comprise T lymphocytes.

75 . The method of claim 72 , wherein the immune effector cells comprise NK cells.

76 . A method of treating a B cell related condition in a subject in need thereof, comprising administering to the subject a therapeutically effect amount of the composition of claim 70 .

77 . The method of claim 76 , wherein the B cell related condition is multiple myeloma, non-Hodgkin's lymphoma, B cell proliferations of uncertain malignant potential, lymphomatoid granulomatosis, post-transplant lymphoproliferative disorder, an immunoregulatory disorder, rheumatoid arthritis, myasthenia gravis, idiopathic thrombocytopenia purpura, anti-phospholipid syndrome, Chagas' disease, Grave's disease, Wegener's granulomatosis, poly-arteritis nodosa, Sjogren's syndrome, pemphigus vulgaris, scleroderma, multiple sclerosis, anti-phospholipid syndrome, ANCA associated vasculitis, Goodpasture's disease, Kawasaki disease, autoimmune hemolytic anemia, and rapidly progressive glomerulonephritis, heavy-chain disease, primary or immunocyte-associated amyloidosis, or monoclonal gammopathy of undetermined significance.

78 . The method of claim 76 , wherein the B cell related condition is a B cell malignancy.

79 . The method of claim 78 , wherein the B cell malignancy is multiple myeloma (MM) or non-Hodgkin's lymphoma (NHL).

80 . The method of claim 79 , wherein the MM is selected from the group consisting of: overt multiple myeloma, smoldering multiple myeloma, plasma cell leukemia, non-secretory myeloma, IgD myeloma, osteosclerotic myeloma, solitary plasmacytoma of bone, and extramedullary plasmacytoma.

81 . The method of claim 79 , wherein the NHL is selected from the group consisting of: Burkitt lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), diffuse large B-cell lymphoma, follicular lymphoma, immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, and mantle cell lymphoma.

82 . The method of claim 76 , wherein the B cell related condition is a plasma cell malignancy.

83 . The method of claim 76 , wherein the B cell related condition is an autoimmune disease.

84 . The method of claim 83 , wherein the autoimmune disease is systemic lupus erythematosus.

85 . The method of claim 83 , wherein the B cell related condition is rheumatoid arthritis.

86 . The method of claim 83 , wherein the B cell related condition is idiopathic thrombocytopenia purpura, or myasthenia gravis, or autoimmune hemolytic anemia.