US2021095001A1PendingUtilityA1

Compositions and methods of phospholipase a2 receptor chimeric autoantibody receptor t cells

59
Assignee: UNIV PENNSYLVANIAPriority: May 2, 2018Filed: May 2, 2019Published: Apr 1, 2021
Est. expiryMay 2, 2038(~11.8 yrs left)· nominal 20-yr term from priority
Inventors:Saar GillJonathan HoganAimee S. PayneBaomei Wang
A61K 40/11A61K 40/32A61K 40/4202A61K 40/31A61K 35/17A61K 2300/00A61K 2121/00C12N 5/0636C07K 2317/34C12N 2740/16043C12N 2510/00C07K 2319/70C07K 2319/40C07K 2319/03C07K 14/70578C07K 14/7056C07K 14/70517C07K 14/7051C07K 14/70503A61P 37/06A61P 13/12A61K 48/005A61K 39/0008A61K 45/06C12N 15/85C07K 14/705C07K 2319/02A61K 48/00C12N 15/62A61K 39/0005
59
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

The invention includes compositions comprising at least one chimeric autoantibody receptor (CAAR) specific for an anti-phospholipase A2 receptor (PLA2R) autoantibody-based B cell receptor, polynucleotides encoding the CAAR, vectors comprising a polynucleotide encoding the CAAR, and recombinant T cells comprising the CAAR. The invention also includes methods of making a genetically modified cell, e.g., a genetically modified T cell, expressing a PLA2R-CAAR wherein the expressed CAAR comprises a PLA2R extracellular domain.

Claims

exact text as granted — not AI-modified
1 . A polynucleotide encoding a chimeric autoantibody receptor (CAAR), wherein the CAAR comprises a phospholipase A2 receptor (PLA2R) autoantigen or fragment thereof, and optionally, a transmembrane domain, an intracellular domain of a costimulatory molecule, and/or a signaling domain. 
     
     
         2 . The polynucleotide of  claim 1 , wherein the PLA2R autoantigen or fragment thereof is selected from the group consisting of:
 (a) an extracellular domain comprising a CysR or ricin B type lectin domain, a fibronectin type II domain, a C-type lectin domain 1 and a C-type lectin domain 2; and,   (b) an extracellular domain comprising a CysR or ricin B type lectin domain, a fibronectin type II domain, a C-type lectin domain 1, a C-type lectin domain 2 and a C-type lectin domain 3.   
     
     
         3 . The polynucleotide of  claim 2 , wherein
 the PLA2R extracellular domain (a) is encoded by a nucleic acid sequence comprising SEQ ID NO: 8, or   the PLA2R extracellular domain (b) is encoded by a nucleic acid sequence comprising SEQ ID NO: 15.   
     
     
         4 . The polynucleotide of  claim 1 , wherein the PLA2R autoantigen or fragment thereof is selected from the group consisting of:
 (a) an extracellular domain comprising a cysteine rich domain,   (b) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, and a C-type lectin domain 1,   (c) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, a C-type lectin domain 1, a C-type lectin domain 2, and a C-type lectin domain 3,   (d) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, a C-type lectin domain 1, a C-type lectin domain 2, a C-type lectin domain 3, and a C-type lectin domain 7,   (e) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, a C-type lectin domain 1, and a C-type lectin domain 7, and   (f) an extracellular domain comprising a cysteine rich domain, a C-type lectin domain 1, and a C-type lectin domain 7.   
     
     
         5 . The polynucleotide of  claim 4 , wherein
 the PLA2R extracellular domain (a) is encoded by a nucleic acid sequence comprising SEQ ID NO: 47 or SEQ ID NO: 60,   the PLA2R extracellular domain (b) is encoded by a nucleic acid sequence comprising SEQ ID NO: 50 or SEQ ID NO: 62,   the PLA2R extracellular domain (c) is encoded by a nucleic acid sequence comprising SEQ ID NO: 52,   the PLA2R extracellular domain (d) is encoded by a nucleic acid sequence comprising SEQ ID NO: 54,   the PLA2R extracellular domain (e) is encoded by a nucleic acid sequence comprising SEQ ID NO: 56, or   the PLA2R extracellular domain (f) is encoded by a nucleic acid sequence comprising SEQ ID NO: 58.   
     
     
         6 . The polynucleotide of  claim 1 , wherein the transmembrane domain comprises a CD8 alpha chain transmembrane domain. 
     
     
         7 . The polynucleotide of  claim 6 , wherein the CD8 alpha chain transmembrane domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 20. 
     
     
         8 . The polynucleotide of  claim 6 , wherein the CD8 alpha chain transmembrane domain comprises the amino acid sequence of SEQ ID NO: 19. 
     
     
         9 . The polynucleotide of  claim 1 , wherein the CAAR further comprises a hinge domain. 
     
     
         10 . The polynucleotide of  claim 9 , wherein the hinge domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 10 or SEQ ID NO: 42 or SEQ ID NO: 64. 
     
     
         11 . The polynucleotide of  claim 9 , wherein the hinge domain comprises the amino acid sequence of SEQ ID NO: SEQ ID NO: 43 or SEQ ID NO: 44. 
     
     
         12 . The polynucleotide of  claim 1 , wherein the CAAR further comprises a GS linker. 
     
     
         13 . The polynucleotide of  claim 12 , wherein the GS linker is encoded by a nucleic acid sequence comprising SEQ ID NO: 68. 
     
     
         14 . The polynucleotide of  claim 12 , wherein the GS linker comprises SEQ ID NO: 69. 
     
     
         15 . The polynucleotide of  claim 1 , wherein the intracellular domain of a costimulatory molecule comprises 4-1BB. 
     
     
         16 . The polynucleotide of  claim 15 , wherein the 4-1BB intracellular domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 22 or SEQ ID NO: 66. 
     
     
         17 . The polynucleotide of  claim 15 , wherein the 4-1BB intracellular domain comprises the amino acid sequence of SEQ ID NO: 21. 
     
     
         18 . The polynucleotide of  claim 1 , wherein the signaling domain comprises a CD3 zeta signaling domain. 
     
     
         19 . The polynucleotide of  claim 18 , wherein the CD3 zeta signaling domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 24 or SEQ ID NO: 72. 
     
     
         20 . The polynucleotide of  claim 18 , wherein the CD3 zeta signaling domain comprises an amino acid sequence of SEQ ID NO: 23 or SEQ ID NO: 45. 
     
     
         21 . The polynucleotide of  claim 1 , wherein the CAAR is encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11, 17, 25, 27, 29, 31, 33, 35, 37, and 39. 
     
     
         22 . The polynucleotide of  claim 1 , wherein the CAAR comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 18, 26, 28, 30, 32, 34, 36, 38, and 40. 
     
     
         23 . The polynucleotide of  claim 1 , wherein the CAAR comprises a phospholipase A2 receptor (PLA2R) autoantigen or fragment thereof, a killer immunoglobulin-like receptor (KIR) transmembrane domain and a KIR cytoplasmic domain, optionally wherein the KIR is KIRS2 or KIR2DS2. 
     
     
         24 . A vector comprising the polynucleotide of  claim 1 . 
     
     
         25 . The vector of  claim 24 , wherein the vector is a lentiviral vector. 
     
     
         26 . The vector of  claim 24 , wherein the vector is a RNA vector. 
     
     
         27 . (canceled) 
     
     
         28 . A chimeric autoantibody receptor (CAAR) comprising an extracellular domain comprising a phospholipase A2 receptor (PLA2R) autoantigen or fragment thereof, and optionally, a transmembrane domain, an intracellular domain of a costimulatory molecule, and/or a signaling domain. 
     
     
         29 . The CAAR of  claim 28 , wherein the PLA2R autoantigen or fragment thereof is selected from the group consisting of:
 (a) an extracellular domain comprising a ricin B type lectin domain, a fibronectin type II domain, a C-type lectin domain 1 and a C-type lectin domain 2; and,   (b) an extracellular domain comprising a ricin B type lectin domain, a fibronectin type II domain, a C-type lectin domain 1, a C-type lectin domain 2 and a C-type lectin domain 3.   
     
     
         30 . The CAAR of  claim 29 , wherein
 the PLA2R extracellular domain (a) is encoded by a nucleic acid sequence comprising SEQ ID NO: 8, or   the PLA2R extracellular domain (b) is encoded by a nucleic acid sequence comprising SEQ ID NO: 15.   
     
     
         31 . The CAAR of  claim 28 , wherein the PLA2R autoantigen or fragment thereof is selected from the group consisting of:
 (a) an extracellular domain comprising a cysteine rich domain,   (b) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, and a C-type lectin domain 1,   (c) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, a C-type lectin domain 1, a C-type lectin domain 2, a C-type lectin domain 3, and a C-type lectin domain 7,   (d) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, a C-type lectin domain 1, a C-type lectin domain 2, a C-type lectin domain 3, and a C-type lectin domain 7,   (e) an extracellular domain comprising a cysteine rich domain, a fibronectin type II domain, a C-type lectin domain 1, and a C-type lectin domain 7, and   (f) an extracellular domain comprising a cysteine rich domain, a C-type lectin domain 1, and a C-type lectin domain 7.   
     
     
         32 . The CAAR of  claim 31 , wherein
 the PLA2R extracellular domain (a) comprises SEQ ID NO: 49 or SEQ ID NO: 61,   the PLA2R extracellular domain (b) comprises the amino acid sequence of SEQ ID NO: 51 or SEQ ID NO: 63,   the PLA2R extracellular domain (c) comprises the amino acid sequence of SEQ ID NO: 53,   the PLA2R extracellular domain (d) comprises the amino acid sequence of SEQ ID NO: 55, or   the PLA2R extracellular domain (e) comprises the amino acid sequence of SEQ ID NO: 57, or   the PLA2R extracellular domain (f) comprises the amino acid sequence of SEQ ID NO: 59.   
     
     
         33 . The CAAR of  claim 28 , wherein the transmembrane domain comprises a CD8 alpha chain transmembrane domain. 
     
     
         34 . The CAAR of  claim 33 , wherein the CD8 alpha chain transmembrane domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 20. 
     
     
         35 . The CAAR of  claim 33 , wherein the CD8 alpha chain transmembrane domain comprises the amino acid sequence of SEQ ID NO: 19. 
     
     
         36 . The CAAR of  claim 28 , wherein the CAAR further comprises a hinge domain. 
     
     
         37 . The CAAR of  claim 36 , wherein the hinge domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 10 or SEQ ID NO: 42 or SEQ ID NO: 64. 
     
     
         38 . The CAAR of  claim 36 , wherein the hinge domain comprises the amino acid sequence of SEQ ID NO: SEQ ID NO: 43 or SEQ ID NO: 44. 
     
     
         39 . The CAAR of  claim 28 , wherein the CAAR further comprises a GS linker. 
     
     
         40 . The CAAR of  claim 39 , wherein the GS linker is encoded by a nucleic acid sequence comprising SEQ ID NO: 68. 
     
     
         41 . The CAAR of  claim 39 , wherein the GS linker comprises SEQ ID NO: 69. 
     
     
         42 . The CAAR of  claim 28 , wherein the intracellular domain of a costimulatory molecule comprises 4-1BB. 
     
     
         43 . The CAAR of  claim 42 , wherein the 4-1BB intracellular domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 22 or SEQ ID NO: 66. 
     
     
         44 . The CAAR of  claim 42 , wherein the 4-1BB intracellular domain comprises the amino acid sequence of SEQ ID NO: 21. 
     
     
         45 . The CAAR of  claim 28 , wherein the signaling domain comprises a CD3 zeta signaling domain. 
     
     
         46 . The CAAR of  claim 45 , wherein the CD3 zeta signaling domain is encoded by a nucleic acid sequence comprising SEQ ID NO: 24 or SEQ ID NO: 72. 
     
     
         47 . The CAAR of  claim 45 , wherein the CD3 zeta signaling domain comprises the amino acid sequence of SEQ ID NO: 23 or SEQ ID NO: 45. 
     
     
         48 . The CAAR of  claim 28 , wherein the CAAR is encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 11, 17, 25, 27, 29, 31, 33, 35, 37, and 39. 
     
     
         49 . The CAAR of  claim 28 , wherein the CAAR comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 18, 26, 28, 30, 32, 34, 36, 38, and 40. 
     
     
         50 . The CAAR of  claim 28 , wherein the CAAR comprises an extracellular domain comprising phospholipase A2 receptor (PLA2R) autoantigen or fragment thereof, a killer immunoglobulin-like receptor (KIR) transmembrane domain and a KIR cytoplasmic domain, optionally wherein the KIR is KIRS2 or KIR2DS2. 
     
     
         51 . A genetically modified cell comprising the CAAR of  claim 28 . 
     
     
         52 . The cell of  claim 51 , wherein the cell expresses the CAAR and has high affinity to autoantibody-based BCRs on B cells. 
     
     
         53 . The cell of  claim 51 , wherein the cell expresses the CAAR and induces killing of B cells expressing autoantibodies. 
     
     
         54 . The cell of  claim 51 , wherein the cell expresses the CAAR and has limited toxicity toward healthy cells. 
     
     
         55 . The cell of  claim 51 , wherein the cell is selected from the group consisting of a helper T cell, a cytotoxic T cell, a memory T cell, regulatory T cell, gamma delta T cell, a natural killer cell, a cytokine induced killer cell, a cell line thereof, a T memory stem cell, a T cell derived from a pluripotent stem and other effector cell. 
     
     
         56 . The cell of  claim 51 , wherein the CAAR comprises an extracellular domain comprising a phospholipase A2 receptor (PLA2R) autoantigen or fragment thereof, a killer immunoglobulin-like receptor (KIR) transmembrane domain and a KIR cytoplasmic domain, optionally wherein the KIR is KIRS2 or KIR2DS2; and wherein the cell further comprises DAP12. 
     
     
         57 . A pharmaceutical composition comprising the polynucleotide of  claim 1 , and a pharmaceutically acceptable excipient. 
     
     
         58 . A method for (a) preventing glomerulus damage in a subject at risk of developing an autoantibody-mediated kidney disease, (b) reducing glomerulus damage in a subject suffering from an autoantibody-mediated kidney disease, and/or (c) treating an autoantibody-mediated kidney disease in a subject, the method comprising: administering to the subject an effective amount of a genetically modified T cell comprising the polynucleotide of  claim 1 . 
     
     
         59 . (canceled) 
     
     
         60 . The method of  claim 58 , wherein the autoantibody mediated kidney disease is selected from the group consisting of a glomerular disease and a primary membranous nephropathy. 
     
     
         61 . The method of  claim 58 , wherein the subject is a human. 
     
     
         62 . The method of  claim 58 , wherein the modified T cell targets B cells. 
     
     
         63 . The method of  claim 58 , wherein the CAAR comprises an extracellular domain comprising phospholipase A2 receptor (PLA2R) autoantigen or fragment thereof, a killer immunoglobulin-like receptor (KIR) transmembrane domain and a KIR cytoplasmic domain, optionally wherein the KIR is KIRS2 or KIR2DS2; and wherein the cell further comprises a polynucleotide encoding DAP12. 
     
     
         64 . (canceled) 
     
     
         65 . (canceled) 
     
     
         66 . (canceled) 
     
     
         67 . (canceled) 
     
     
         68 . The vector of  claim 24 , wherein the vector comprises an inducible promoter operably linked to the polynucleotide encoding the CAAR. 
     
     
         69 . The cell of  claim 51 , wherein the cell comprises a polynucleotide encoding the CAAR operably linked to an inducible promoter. 
     
     
         70 . A pharmaceutical composition comprising the CAAR of  claim 28  and a pharmaceutically acceptable excipient. 
     
     
         71 . A pharmaceutical composition comprising the cell of  claim 51 , and a pharmaceutically acceptable excipient.

Join the waitlist — get patent alerts

Track US2021095001A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.