Engineered phagocytic receptor compositions and methods of use thereof

Abstract

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 therapeutically effective amount of a therapeutic agent, wherein the therapeutic agent comprises a nanoparticle delivery vehicle comprising a recombinant polynucleic acid, wherein the recombinant polynucleic acid comprises a sequence encoding a chimeric fusion protein (CFP), the CFP comprising:   (i) an extracellular domain comprising an antigen binding domain, and   (ii) a transmembrane domain operatively linked to the extracellular domain;   wherein the transmembrane domain is a transmembrane domain from a protein that dimerizes with endogenous FcR-gamma receptors in myeloid cells; wherein after administration of the pharmaceutical composition to a human subject the CFP is expressed on the surface of myeloid cells of the human subject.   
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein the antigen binding domain is a CD5 binding domain that comprises Fab fragment, an scFv domain or an sdAb domain. 
     
     
         3 . The pharmaceutical composition of  claim 2 , wherein the CD5 binding domain comprises an scFv comprising: (i) a variable heavy chain (V H ) sequence with at least 90% sequence identity to SEQ ID NO: 1; and (ii) a variable light chain (V L ) sequence with at least 90% sequence identity to SEQ ID NO: 2. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein the CFP further comprises an intracellular domain, wherein the intracellular domain comprises one or more intracellular signaling domains, and wherein the one or more intracellular signaling domains comprises an intracellular signaling domain from FcγR, FcαR, FcεR or CD3zeta. 
     
     
         5 . The pharmaceutical composition of  claim 4 , wherein the intracellular signaling domain comprises a phosphoinositide 3-kinase (PI3K) recruitment domain. 
     
     
         6 . The pharmaceutical composition of  claim 5 , wherein the PI3K recruitment domain binds a p85 regulatory subunit of PI3K, and wherein the PI3K recruitment domain is from CD19. 
     
     
         7 . The pharmaceutical composition of  claim 4 , wherein the intracellular domain further comprises one or more additional intracellular signaling domains. 
     
     
         8 . The pharmaceutical composition of  claim 4 , wherein the intracellular domain comprises an intracellular signaling domain with at least 90% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 27. 
     
     
         9 . The pharmaceutical composition of  claim 1 , wherein the antigen binding domain is a HER2 binding domain that comprises a Fab fragment, an scFv domain or an sdAb domain 
     
     
         10 . The pharmaceutical composition of  claim 9 , wherein the HER2 binding domain comprises an scFv comprising: (i) a variable heavy chain V H  sequence with at least 90% sequence identity to SEQ ID NO: 8; and (ii) a V L  sequence with at least 90% sequence identity to SEQ ID NO: 9. 
     
     
         11 . The pharmaceutical composition of  claim 5 , wherein the PI3K recruitment domain domain comprises a sequence with at least 90% sequence identity to SEQ ID NO: 4. 
     
     
         12 . The pharmaceutical composition of  claim 7 , wherein the one or more additional intracellular signaling domains comprise an intracellular signaling domain from an intracellular signaling domain of CD40. 
     
     
         13 . The pharmaceutical composition of  claim 12 , wherein the intracellular signaling domain of CD40 comprises a sequence with at least 90% sequence identity to SEQ ID NO: 5. 
     
     
         14 . The pharmaceutical composition of  claim 1 , wherein the transmembrane domain comprises a transmembrane domain from CD16a, CD64, CD68 or CD89. 
     
     
         15 . (canceled) 
     
     
         16 . 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 antigen binding domain. 
     
     
         17 . The pharmaceutical composition of  claim 16 , wherein the extracellular domain comprises a sequence with at least 90% sequence identity to SEQ ID NO: 7. 
     
     
         18 . The pharmaceutical composition of  claim 1 , wherein the CFP comprises a sequence having at least 90% sequence identity to SEQ ID NO: 14. 
     
     
         19 . The pharmaceutical composition of  claim 1 , wherein the recombinant polynucleic acid is an mRNA. 
     
     
         20 . (canceled) 
     
     
         21 . The pharmaceutical composition of  claim 1 , wherein the nanoparticle delivery vehicle comprises a lipid nanoparticle. 
     
     
         22 . The pharmaceutical composition of  claim 21 , wherein the recombinant polynucleic acid is an mRNA and the lipid nanoparticle encapsulates the mRNA. 
     
     
         23 . The pharmaceutical composition of  claim 21 , wherein the lipid nanoparticle comprises a polar lipid and a non-polar lipid. 
     
     
         24 . The pharmaceutical composition of  claim 21 , wherein the lipid nanoparticle is from 100 to 300 nm in diameter. 
     
     
         25 . The pharmaceutical composition of  claim 1 , wherein the antigen binding domain is a GPC3 binding domain that comprises a Fab fragment, an scFv domain or an sdAb domain. 
     
     
         26 . The pharmaceutical composition of  claim 1 , wherein the extracellular domain is an extracellular domain from CD16a, CD64, CD68 or CD89. 
     
     
         27 . The pharmaceutical composition of  claim 26 , wherein the transmembrane domain is a transmembrane domain from CD16a, CD64, CD68 or CD89. 
     
     
         28 . The pharmaceutical composition of  claim 27 , wherein the CFP further comprises an intracellular domain, wherein the intracellular domain comprises an intracellular domain from CD16a, CD64, CD68 or CD89. 
     
     
         29 . A method of treating a cancer in a human subject in need thereof, comprising administering to the human subject a pharmaceutical composition of  claim 1 . 
     
     
         30 . The method of  claim 29 , wherein the cancer is a solid tumor. 
     
     
         31 . The method of  claim 29 , wherein the cancer is a lymphoma. 
     
     
         32 . The method of  claim 29 , wherein the cancer is T cell lymphoma.