Compositions and methods of phospholipase A2 receptor chimeric autoantibody receptor T cells
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.
1. An isolated polynucleotide encoding a chimeric autoantibody receptor (CAAR), wherein the CAAR comprises:
(i) an extracellular domain comprising a phospholipase A2 receptor (PLA2R) autoantigen comprising the amino acid sequence of SEQ ID NO: 59;
(ii) a CD8 alpha chain transmembrane domain;
(iii) a 4-1BB intracellular domain; and
(iv) a CD3 zeta signaling domain.
2. The polynucleotide of claim 1 , wherein the CD8 alpha chain transmembrane domain comprises the amino acid sequence of SEQ ID NO: 19.
3. The polynucleotide of claim 1 , wherein the 4-1BB intracellular domain comprises the amino acid sequence of SEQ ID NO: 21.
4. The polynucleotide of claim 1 , wherein the CD3 zeta signaling domain comprises the amino acid sequence of SEQ ID NO: 23 or SEQ ID NO: 45.
5. The polynucleotide of claim 1 , wherein:
(i) the CD8 alpha chain transmembrane domain comprises the amino acid sequence of SEQ ID NO: 19;
(ii) the 4-1BB intracellular domain comprises the amino acid sequence of SEQ ID NO: 21; and
(iii) CD3 zeta signaling domain comprises the amino acid sequence of SEQ ID NO: 45.
6. A recombinant vector comprising the polynucleotide of claim 1 .
7. A chimeric autoantibody receptor (CAAR) comprising:
(i) an extracellular domain comprising a phospholipase A2 receptor (PLA2R) autoantigen comprising the amino acid sequence of SEQ ID NO: 59;
(ii) a CD8 alpha chain transmembrane domain;
(iii) a 4-1BB intracellular domain; and
(iv) a CD3 zeta signaling domain.
8. The CAAR of claim 7 , wherein the CD8 alpha chain transmembrane domain comprises the amino acid sequence of SEQ ID NO: 19.
9. The CAAR of claim 7 , wherein the 4-1BB intracellular domain comprises the amino acid sequence of SEQ ID NO: 21.
10. The CAAR of claim 7 , wherein the CD3 zeta signaling domain comprises the amino acid sequence of SEQ ID NO: 23 or SEQ ID NO: 45.
11. The CAAR of claim 7 , wherein:
(i) the CD8 alpha chain transmembrane domain comprises the amino acid sequence of SEQ ID NO: 19;
(ii) the 4-1BB intracellular domain comprises the amino acid sequence of SEQ ID NO: 21; and
(iii) the CD3 zeta signaling domain comprises the amino acid sequence of SEQ ID NO: 45.
12. A genetically modified cell comprising thy: CAAR of claim 7 .
13. A pharmaceutical composition comprising the polynucleotide of claim 1 and a pharmaceutically acceptable excipient.
14. A pharmaceutical composition comprising the genetically modified cell of claim 12 and a pharmaceutically acceptable excipient.
15. A method for treating an autoantibody-mediated kidney disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the genetically modified cell of claim 12 , thereby treating the autoantibody mediated kidney disease in the subject.
16. The method of claim 15 , wherein the autoantibody mediated kidney disease is selected from the group consisting of a glomerular disease and a primary membranous nephropathy.
17. A method for reducing glomerulus damage in a subject at risk of or suffering from an autoantibody-mediated kidney disease, the method comprising administering to the subject an effective amount of the genetically modified cell of claim 12 , thereby preventing or reducing glomerulus damage in the subject.
18. The method of claim 17 , wherein the autoantibody mediated kidney disease is selected from the group consisting of a glomerular disease and a primary membranous nephropathy.