Bcma chimeric antigen receptors and uses thereof

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.

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, wherein the anti-BCMA binding domain comprises a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HC CDR1), a heavy chain complementarity determining region 2 (HC CDR2), and a heavy chain complementarity determining region 3 (HC CDR3), and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LC CDR1), a light chain complementarity determining region 2 (LC CDR2), and a light chain complementarity determining region 3 (LC CDR3), wherein the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of:
 (i) SEQ ID NOs: 44, 45, 84, 54, 55, and 56, respectively; or   (ii) SEQ ID NOs: 179, 180, 181, 147, 182, and 183, respectively.   
     
     
         2 . An isolated CAR comprising an anti-BCMA binding domain, a transmembrane domain, and an intracellular signaling domain, wherein the anti-BCMA binding domain comprises a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HC CDR1), a heavy chain complementarity determining region 2 (HC CDR2), and a heavy chain complementarity determining region 3 (HC CDR3), a light chain complementarity determining region 1 (LC CDR1), a light chain complementarity determining region 2 (LC CDR2), and a light chain complementarity determining region 3 (LC CDR3), wherein the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of:
 (i) SEQ ID NOs: 44, 45, 84, 54, 55, and 56, respectively; or   (ii) SEQ ID NOs: 179, 180, 181, 147, 182, and 183, respectively.   
     
     
         3 . (canceled) 
     
     
         4 . The isolated nucleic acid molecule of  claim 1 , wherein the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of:
 (i) SEQ ID NOs: 44, 45, 76, 54, 55, and 56, respectively:   (ii) SEQ ID NOs: 44, 45, 46, 54, 55, and 56, respectively:   (iii) SEQ ID NOs: 44, 45, 68, 54, 55, and 56, respectively;   (iv) SEQ ID NOs: 137, 138, 139, 147, 148, and 149, respectively; or   (v) SEQ ID NOs: 160, 161, 162, 147, 170, and 171, respectively.   
     
     
         5 . The isolated nucleic acid molecule of  claim 1 , wherein the VH comprises the amino acid sequence of SEQ ID NO: 78, 52, 70, 145, or 168, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         6 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 79, 53, 71, 146, or 169, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         7 . The isolated nucleic acid molecule of  claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NO: 61, 154, or 173, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         8 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 62, 155, or 174, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         9 . The isolated nucleic acid molecule of  claim 1 , wherein the VH and VL comprise the amino acid sequences of:
 (i) SEQ ID NOs: 78 and 61, respectively:   (ii) SEQ ID NOs: 52 and 61, respectively;   (iii) SEQ ID NOs: 70 and 61, respectively:   (iv) SEQ ID NOs: 145 and 154, respectively, or   (v) SEQ ID NOs: 168 and 173, respectively.   
     
     
         10 . The isolated nucleic acid molecule of  claim 1 , wherein the anti-BCMA binding domain comprises a single-chain fragment variable (scFv) comprising the amino acid sequence of SEQ ID NO: 80, 64, 72, 156, or 175, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         11 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 81, 65, 73, 157, or 176, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         12 . The isolated nucleic acid molecule of  claim 1 , wherein the CAR comprises the amino acid sequence of SEQ ID NO: 82, 66, 74, 158, or 177, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         13 . The isolated nucleic acid molecule of  claim 1 , wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 83, 67, 75, 159, or 178, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto. 
     
     
         14 .- 35 . (canceled) 
     
     
         36 . The isolated nucleic acid molecule of  claim 1 , wherein the VH and VL are connected by a linker, comprising the amino acid sequence of SEQ ID NO: 63 or 104. 
     
     
         37 . The isolated nucleic acid molecule of  claim 1 , wherein:
 (i) the transmembrane domain comprises a transmembrane domain of a protein chosen from the alpha, beta or zeta chain of T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 or CD154;   (ii) the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 6, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or   (iii) the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 17, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.   
     
     
         38 . The isolated nucleic acid molecule of  claim 1 , wherein the anti-BCMA binding domain is connected to the transmembrane domain by a hinge region, wherein:
 (i) the hinge region comprises the amino acid sequence of SEQ ID NO: 2, 3, or 4, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or   (ii) the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 13, 14, or 15, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.   
     
     
         39 . The isolated nucleic acid molecule of  claim 1 , wherein the intracellular signaling domain comprises a primary signaling domain, wherein:
 (i) the primary signaling domain comprises a functional signaling domain derived from CD3 zeta, TCR zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, CD278 (ICOS), FcεRI, DAP10, DAP12, or CD66d;   (ii) the primary signaling domain comprises the amino acid sequence of SEQ ID NO: 9 or 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or   (iii) the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, or SEQ ID NO: 256, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.   
     
     
         40 . The isolated nucleic acid molecule of  claim 1 , wherein the intracellular signaling domain comprises a costimulatory signaling domain, wherein:
 (i) the costimulatory signaling domain comprises a functional signaling domain derived from a MHC class I molecule, a TNF receptor protein, an Immunoglobulin-like protein, a cytokine receptor, an integrin, a signalling lymphocytic activation molecule (SLAM protein), an activating NK cell receptor, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, ICAM-1, 4-1BB (CD137), B7-H3, 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, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAMI, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMFI, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG/Cbp, CD19a, CD28-OX40, CD28-4-1BB, or a ligand that specifically binds with CD83;   (ii) the costimulatory signaling domain comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or   (iii) the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 18 or SEQ ID NO: 255, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.   
     
     
         41 . The isolated nucleic acid molecule of  claim 1 , wherein the intracellular signaling domain comprises a functional signaling domain derived from 4-1BB and a functional signaling domain derived from CD3 zeta, optionally wherein the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 7 (or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto) and the amino acid sequence of SEQ ID NO: 9 or 10 (or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto), optionally wherein the intracellular signaling domain comprises the amino acid sequence of SEQ ID NO: 7 and the amino acid sequence of SEQ ID NO: 9 or 10. 
     
     
         42 . The isolated nucleic acid molecule of  claim 1 , wherein the CAR further comprises a leader sequence comprising the amino acid sequence of SEQ ID NO: 1. 
     
     
         43 . The isolated nucleic acid molecule of  claim 1 , wherein the CAR comprises one or more of the following properties:
 (i) the CAR, when expressed in a cell, activates NFAT signaling in the cell in the presence of BCMA-expressing cells;   (ii) the CAR, when expressed in a cell, induces cytotoxicity of BCMA-expressing cells; and   (iii) the CAR, when expressed in a cell, induces expression of a cytokine in the cell in the presence of BCMA-expressing cells.   
     
     
         44 . (canceled) 
     
     
         45 . An anti-BCMA binding domain comprising a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HC CDR1), a heavy chain complementarity determining region 2 (HC CDR2), and a heavy chain complementarity determining region 3 (HC CDR3), and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LC CDR1), a light chain complementarity determining region 2 (LC CDR2), and a light chain complementarity determining region 3 (LC CDR3), wherein the HC CDR1, HC CDR2, HC CDR3, LC CDR1, LC CDR2, and LC CDR3 comprise the amino acid sequences of:
 (i) SEQ ID NOs: 44, 45, 84, 54, 55, and 56, respectively; or   (ii) SEQ ID NOs: 179, 180, 181, 147, 182, and 183, respectively.   
     
     
         46 .- 47 . (canceled) 
     
     
         48 . A vector comprising the nucleic acid molecule of  claim 1 . 
     
     
         49 . (canceled) 
     
     
         50 . A cell comprising the nucleic acid molecule of  claim 1 . 
     
     
         51 . A method of making a cell comprising transducing a cell with the vector of  claim 48 . 
     
     
         52 . A method of making an RNA-engineered cell comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, wherein the RNA comprises the nucleic acid molecule of  claim 1 . 
     
     
         53 . A method of providing an anti-tumor immunity in a subject comprising administering to the subject an effective amount of the cell of  claim 50 . 
     
     
         54 . A method of treating a subject having a disease associated with expression of BCMA comprising administering to the subject an effective amount of the cell of  claim 50 . 
     
     
         55 .- 60 . (canceled)