Engineered phagocytic receptor compositions and methods of use thereof
The present disclosure provides compositions and methods for making and using engineered killer phagocytic cells for immunotherapy in cancer or infection by expressing a chimeric antigen receptor having an enhanced phagocytic activity, the chimeric receptor is encoded by a recombinant nucleic acid.
1. A pharmaceutical composition comprising:
(a) myeloid cells from a human subject comprising a recombinant polynucleic acid, wherein the myeloid cells are CD14 + and CD16 − , and wherein the recombinant polynucleic acid comprises a sequence encoding a chimeric fusion protein (CFP), the CFP comprising:
(i) an extracellular domain comprising a CD5 binding domain, wherein the CD5 binding domain comprises an scFv comprising a variable heavy chain (VH) sequence with SEQ ID NO: 1 and a variable light chain (VL) sequence with SEQ ID NO: 2,
(ii) a CD8 transmembrane domain operatively linked to the extracellular domain; and
(iii) an intracellular domain comprising at least two intracellular signaling domains, wherein the at least two intracellular signaling domains comprise:
(A) a first intracellular signaling domain derived from FcγR or FcεR, and
(B) a second intracellular signaling domain comprising a PI3K recruitment domain; and
(b) a pharmaceutically acceptable carrier;
wherein the myeloid cells express the CFP and exhibit at least a 1.1 fold increase in phagocytosis of a target cell expressing CD5; and
wherein the pharmaceutical composition comprises an effective amount of the myeloid cells to inhibit growth of a solid tumor when administered to a human subject with the solid tumor.
2. The pharmaceutical composition of claim 1 , wherein the CD5 binding domain binds to CD5 with an affinity of 250 nM or less.
3. The pharmaceutical composition of claim 1 , wherein the intracellular domain further comprises one or more additional intracellular signaling domains.
4. The pharmaceutical composition of claim 3 , wherein the one or more additional intracellular signaling domains comprise an intracellular signaling domain derived from a receptor other than Megf10, MerTk, FcαR, and Bai1.
5. The pharmaceutical composition of claim 3 , wherein the one or more additional intracellular signaling domains comprises a proinflammatory signaling domain.
6. The pharmaceutical composition of claim 3 , wherein the one or more additional intracellular signaling domains comprise an intracellular signaling domain derived from an intracellular signaling domain of CD40.
7. The pharmaceutical composition of claim 6 , wherein the intracellular signaling domain derived from an intracellular signaling domain of CD40 comprises a sequence with at least 90% sequence identity to SEQ ID NO: 5.
8. The pharmaceutical composition of claim 1 , wherein the intracellular domain comprises an intracellular signaling domain with at least 90% sequence identity to SEQ ID NO: 3.
9. The pharmaceutical composition of claim 1 , wherein the PI3K recruitment domain comprises a sequence with at least 90% sequence identity to SEQ ID NO: 4.
10. The pharmaceutical composition of claim 1 , wherein the CD8 transmembrane domain comprises a sequence with at least 90% sequence identity to SEQ ID NO: 6.
11. The pharmaceutical composition of claim 1 , wherein the extracellular domain further comprises a hinge domain derived from CD8, wherein the hinge domain is operatively linked to the transmembrane domain and the CD5 binding domain.
12. The pharmaceutical composition of claim 11 , wherein the hinge domain derived from CD8 comprises a sequence with at least 90% sequence identity to SEQ ID NO: 7.
13. The pharmaceutical composition of claim 1 , wherein the recombinant polynucleic acid is an mRNA or circRNA.
14. The pharmaceutical composition of claim 1 , wherein the CFP comprises a sequence having at least 90% sequence identity to SEQ ID NO: 14.
15. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition comprises at least 6×10 6 of the myeloid cells.
16. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition comprises an effective amount of the myeloid cells to inhibit growth of a solid tumor that is T cell lymphoma when administered to a human subject with the solid tumor that is T cell lymphoma.