Therapeutic cell compositions and methods of manufacturing and 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) an ex vivo population of human cells comprising a recombinant polynucleic acid, wherein the recombinant polynucleic acid comprises a sequence encoding a chimeric fusion protein (CFP), wherein:
(i) at least 50% of the cells in the ex vivo population of human cells are CD14+ and CD16−, and
(ii) less than 10% of the cells in the ex vivo population of human cells are dendritic cells; and
(b) a pharmaceutically acceptable excipient.
2. The pharmaceutical composition of claim 1 , wherein at least 50% of the cells in the ex vivo population of human cells are CCR2+ and/or CCR5+.
3. The pharmaceutical composition of claim 1 , wherein at least 50% of the cells in the ex vivo population of human cells are CD63+.
4. The pharmaceutical composition of claim 1 , wherein at least 50% of the cells in the ex vivo population of human cells are CD56−, CD3−, and/or CD19−.
5. The pharmaceutical composition of claim 1 , wherein less than 40% of the cells in the ex vivo population of human cells are macrophage cells.
6. The pharmaceutical composition of claim 1 , wherein:
(a) at least 50% of the cells in the ex vivo population of human cells are CCR2+ and/or CCR5+;
(b) at least 50% of the cells in the ex vivo population of human cells are CD63+;
(c) at least 50% of the cells in the ex vivo population of human cells are CD56−, CD3−, and/or CD19−; and
(d) less than 40% of the cells in the ex vivo population of human cells are macrophage cells.
7. The pharmaceutical composition of claim 1 , wherein the recombinant polynucleic acid comprises RNA.
8. The pharmaceutical composition of claim 1 , wherein the recombinant polynucleic acid comprises a sequence encoding a CFP, and the ex vivo population of human cells lacks tonic signaling through the CFP.
9. The pharmaceutical composition of claim 1 , wherein the CFP comprises:
(a) an extracellular domain comprising an antigen binding domain, and
(b) a transmembrane domain operatively linked to the extracellular domain.
10. The pharmaceutical composition of claim 9 , wherein the antigen binding domain is a CD5 binding domain or a HER2 binding domain.
11. The pharmaceutical composition of claim 9 , wherein the CFP further comprises an intracellular domain derived from a phagocytic receptor or a scavenger receptor.
12. The pharmaceutical composition of claim 9 , wherein the CFP comprises:
(a) an extracellular domain comprising:
(i) a scFv that specifically binds CD5 or HER2, and
(ii) a hinge domain derived from CD8, or CD28 or an extracellular domain of CD68 or a portion thereof;
(b) a CD8 transmembrane domain, a CD28 transmembrane domain or a CD68 transmembrane domain; and
(c) an intracellular domain comprising at least two intracellular signaling domains, wherein the at least two intracellular signaling domains comprise
(i) a first intracellular signaling domain derived from FcγR or FcεR, and
(ii) a second intracellular signaling domain that:
(A) comprises a PI3-kinase (PI3K) recruitment domain, or
(B) is derived from CD40.
13. A method of treating a disease or condition in a human subject in need thereof comprising: administering the pharmaceutical composition of claim 1 to the human subject.
14. The method of claim 13 , wherein cells of the ex vivo population of human cells:
(a) differentiate into effector cells in the subject after administration;
(b) infiltrate into a diseased site of the subject after administration or migrate to a diseased site of the subject after administration; or
(c) have a life-span of at least 5 days in the subject after administration.
15. The method of claim 13 , wherein the ex vivo population of human cells is from the human subject.
16. The method of claim 13 , wherein the pharmaceutical composition is administered to the human subject within 72 hours after the recombinant polynucleic acid has been introduced into the ex vivo population of human cells.
17. The method of claim 13 , wherein the ex vivo population of human cells has been cultured for less than 48 hours ex vivo prior to administration.
18. A method of negatively selecting cells for preparing the pharmaceutical composition of claim 1 , the method comprising: (a) contacting a biological sample from a human subject with an anti-CD16 antibody and one or more antibodies selected from anti-CD56 antibody, anti-CD3 antibody and anti-CD19 antibody, (b) collecting cells in the biological sample that are not bound by the anti-CD16 antibody and not bound by the one or more antibodies, and (c) introducing a recombinant polynucleic acid comprising a sequence encoding a CFP into cells collected from (b), thereby forming a population of cells, wherein: (i) at least 50% of the cells in the population of cells are CD14+ and CD16, and (ii) less than 10% of the cells in the population of cells are dendritic cells.
19. The method of claim 18 , wherein the method comprises flow cytometry.
20. The pharmaceutical composition of claim 1 , wherein the ex vivo population of human cells comprises at least 1×10 7 cells.
21. The pharmaceutical composition of claim 1 , wherein the antigen binding domain is a cancer antigen binding domain.