Chimeric antigen receptors targeting Fc receptor-like 5 and uses thereof
The presently disclosed subject matter provides for methods and compositions for treating a neoplasia (e.g., multiple myeloma). It relates to chimeric antigen receptors (CARs) that specifically target Fc Receptor-like 5 (FcRL5), e.g., domain 9 of FcRL5, and immunoresponsive cells comprising such CARs. The presently disclosed FcRL5-targeted CARs have enhanced immune-activating properties, including anti-tumor activity.
1. A chimeric antigen receptor (CAR), comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular domain, wherein the extracellular antigen-binding domain specifically binds to Fc Receptor-Like 5 (FcRL5) and comprises:
(a) a light chain variable region comprising a CDR1, a CDR2, and a CDR3 of the light chain variable region sequence set forth in SEQ ID NO:3, and a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable region sequence set forth in SEQ ID NO:4;
(b) a light chain variable region comprising a CDR1, a CDR2, and a CDR3 of the light chain variable region sequence set forth in SEQ ID NO:115, and a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable region sequence set forth in SEQ ID NO:116;
(c) a light chain variable region comprising a CDR1, a CDR2, and a CDR3 of the light chain variable region sequence set forth in SEQ ID NO:219, and a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable region sequence set forth in SEQ ID NO:220; or
(d) a light chain variable region comprising a CDR1, a CDR2, and a CDR3 of the light chain variable region sequence set forth in SEQ ID NO:235, and a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 of the heavy chain variable region sequence set forth in SEQ ID NO: 236.
2. The CAR of claim 1 , wherein:
(a) the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:312, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:313, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 314; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:309, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:310, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 311;
(b) the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:329, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:330, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 443; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:440, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:441, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 442;
(c) the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:533, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:534, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 535; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:403, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:404, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 532; or
(d) the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:544, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:448, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 545; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:411, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:412, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 543.
3. The CAR of claim 1 , wherein:
(a) the light chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:3, and the heavy chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:4;
(b) the light chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:115, and the heavy chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:116;
(c) the light chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:219, and the heavy chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:220; or
(d) the light chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:235, and the heavy chain variable region comprises an amino acid sequence that has at least about 90% sequence identity to the sequence set forth in SEQ ID NO:236.
4. The CAR of claim 3 , wherein:
(a) the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:3, and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:4;
(b) the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:115, and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:116;
(c) the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:219, and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:220; or
(d) the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:235, and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 236.
5. The CAR of claim 2 , wherein the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:533, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:534, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:535; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:403, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:404, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:532.
6. The CAR of claim 2 , wherein the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:544, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:448, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:545; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:411, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:412, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:543.
7. The CAR of claim 2 , wherein the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:329, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:330, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:443; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO:440, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO:441, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO:442.
8. The CAR of claim 4 , wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:219, and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:220.
9. The CAR of claim 4 , wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:235, and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:236.
10. The CAR of claim 4 , wherein the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO:115, and the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:116.
11. The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises a single-chain variable fragment (scFv).
12. The CAR of claim 11 , wherein the extracellular antigen-binding domain comprises a human scFv.
13. The CAR of claim 1 , wherein the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide, or a combination thereof.
14. The CAR of claim 1 , wherein the intracellular domain comprises a CD3ζ polypeptide.
15. The CAR of claim 14 , wherein the intracellular domain further comprises at least one signaling region.
16. The CAR of claim 15 , wherein the at least one signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a PD-1 polypeptide, a CTLA-4 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide, or a combination thereof.
17. The CAR of claim 15 , wherein the at least one signaling region comprises at least one co-stimulatory signaling region.
18. The CAR of claim 17 , wherein the at least one co-stimulatory signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combination thereof.
19. The CAR of claim 1 , wherein:
(a) the transmembrane domain comprises a CD28 polypeptide, and the intracellular domain comprises a CD3ζ polypeptide and a co-stimulatory signaling region that comprises a CD28 polypeptide;
(b) the transmembrane domain comprises a CD8 polypeptide, and the intracellular domain comprises a CD3ζ polypeptide and a co-stimulatory signaling region that comprises a 4-1BB polypeptide; or
(c) the transmembrane domain comprises a CD28 polypeptide, and the intracellular domain comprises a CD3ζ polypeptide and a co-stimulatory signaling region that comprises a 4-1BB polypeptide.
20. An immunoresponsive cell comprising the CAR of claim 1 .
21. The immunoresponsive cell of claim 20 , wherein the immunoresponsive cell is further transduced with a nucleic acid molecule encoding at least one co-stimulatory ligand such that the immunoresponsive cell expresses the at least one co-stimulatory ligand; or a nucleic acid molecule encoding at least one cytokine such that the immunoresponsive cell secretes the at least one cytokine.
22. The immunoresponsive cell of claim 20 , wherein the immunoresponsive cell is selected from the group consisting of a T cell, a Natural Killer (NK) cell, a cytotoxic T lymphocyte (CTL), a regulatory T cell, a human embryonic stem cell, a lymphoid progenitor cell, a T cell-precursor cell, and a pluripotent stem cell from which lymphoid cells may be differentiated.
23. The immunoresponsive cell of claim 20 , wherein the immunoresponsive cell is a T cell.
24. A nucleic acid molecule encoding the CAR of claim 1 .
25. The nucleic acid molecule of claim 24 , comprising the nucleotide sequence set forth in SEQ ID NO:953, SEQ ID NO:957, or SEQ ID NO:958, or a sequence having at least about 90% identity to the nucleotide sequence set forth in SEQ ID NO: 953, SEQ ID NO:957, or SEQ ID NO:958.
26. A vector comprising the nucleic acid molecule of claim 24 .
27. A host cell expressing the nucleic acid molecule of claim 24 .
28. A T cell comprising the CAR of claim 1 .
29. A method for producing an immunoresponsive cell that binds to Fc Receptor-Like 5 (FcRL5), comprising introducing into an immunoresponsive cell a nucleic acid molecule that encodes the CAR of claim 1 .
30. A pharmaceutical composition comprising an effective amount of the immunoresponsive cell of claim 20 and a pharmaceutically acceptable excipient.
31. A kit for treating a tumor, the kit comprising the immunoresponsive cell of claim 20 and written instructions for using the immunoresponsive cell for treating a subject having a tumor.