Chimeric antigen receptors for phagocytosis
The present disclosure generally relate to chimeric antigen receptors (CARs) that bind an engulfment receptor expressed on the surface of a phagocytic cell and activate the endogenous phagocytic signaling pathway. Also disclosed are compositions and methods useful for producing such CARs, nucleic acids encoding same, phagocytic cells that have been modified to include a targeted effector activity directed towards a cell of interest such as, e.g., a cancer cell, as well as for modifying a cell and/or for the treatment of various health disorders such as cancer, including solid tumor and hematologic malignancy.
1. A pharmaceutical composition comprising
(a) a cell comprising a recombinant polynucleotide, wherein the recombinant polynucleotide comprises a sequence encoding a recombinant polypeptide comprising:
(i) an extracellular domain comprising an antigen-binding region specific for a cell surface antigen,
(ii) a transmembrane domain,
(iii) an intracellular domain comprising:
(A) an intracellular signaling domain from FcRγ, CD3zeta or Megf10, and
(B) a p85-recruitment domain that binds a p85 regulatory subunit of phosphoinositide 3-kinase (PI3K), wherein the p85-recruitment domain is from CD19; and
(b) a pharmaceutically acceptable carrier.
2. The pharmaceutical composition of claim 1 , wherein the antigen-binding region comprises an a single-chain variable fragment (scFv) or a single domain antibody (sdAb) variable domain.
3. The pharmaceutical composition of claim 1 , wherein the cell surface antigen is selected from the group consisting of CD19, CD22, HER2 (ERBB2/neu), Mesothelin, PSCA, CD123, CD30, CD171, CD138, CS-1, CLECL1, CD33, CD79b, EGFRvIII, GD2, GD3, BCMA, PSMA, ROR1, FLT3, TAG72, CD38, CD44v6, CEA, EPCAM, B7H3 (CD276), KIT (CD117), CD213A2, IL-1 IRa, PRSS21, VEGFR2, FSHR, TROP2, CD24, MUC-16, PDGFR-beta, SSEA-4, CD20, MUC1, EGFR, NCAM, Prostase, PAP, ELF2M, Ephrin B2, FAP, EphA2, GM3, TEM1/CD248, TEM7R, CLDN6, TSHR, GPRC5D, CD97, CD179a, ALK, and IGLL1.
4. The pharmaceutical composition of claim 1 , wherein the cell is selected from the group consisting of a macrophage, a dendritic cell, a mast cell, a monocyte, a neutrophil, a microglial cell and an astrocyte.
5. The pharmaceutical composition of claim 4 , wherein the cell is a monocyte cell.
6. The pharmaceutical composition of claim 1 , wherein the transmembrane domain comprises a stalk and/or transmembrane domain from CD8, Megf10, FcRγ, Bai1, MerTK, TIM4, Stabilin-1, Stabilin-2, RAGE, CD300f, Integrin subunit αv, Integrin subunit β5, CD36, LRP1, SCARF1, C1Qa, Axl, CD45 or CD86.
7. The pharmaceutical composition of claim 6 , wherein the transmembrane domain comprises a transmembrane domain from CD8.
8. The pharmaceutical composition of claim 1 , wherein the intracellular signaling domain comprises at least 1 ITAM motif.
9. The pharmaceutical composition of claim 1 , wherein binding of the antigen-binding region to the cell surface antigen activates an endogenous phagocytic signaling pathway of the cell.
10. The pharmaceutical composition of claim 1 , wherein the intracellular domain comprises the intracellular signaling domain from FcRγ.
11. The pharmaceutical composition of claim 1 , wherein the intracellular domain comprises the intracellular signaling domain from CD3zeta.
12. The pharmaceutical composition of claim 1 , wherein the intracellular domain comprises the intracellular signaling domain from Megf10.
13. The pharmaceutical composition of claim 1 , wherein the p85-recruitment domain comprises amino acids 345-379 of SEQ ID NO: 4.
14. The pharmaceutical composition of claim 1 , wherein the p85-recruitment domain is operably linked downstream to the intracellular signaling domain from FcRγ.
15. The pharmaceutical composition of claim 1 , wherein the recombinant polypeptide comprises SEQ ID NO: 5.
16. The pharmaceutical composition of claim 1 , wherein the sequence encoding the recombinant polypeptide comprises SEQ ID NO: 14.
17. The pharmaceutical composition of claim 1 , wherein the extracellular domain comprises an scFv specific to CD19, CD22 or HER2 (ERBB2/neu); and wherein the transmembrane domain is a transmembrane domain from CD8.
18. A method of treating cancer in a subject in need thereof comprising administering the pharmaceutical composition of claim 1 to the subject, thereby treating the cancer in the subject.
19. The method of claim 18 , wherein the cancer is a lymphoma.