Modified monocytes/macrophage expressing chimeric antigen receptors and uses thereof
The present invention includes methods and compositions for treating cancer, whether a solid tumor or a hematologic malignancy. By expressing a chimeric antigen receptor in a monocyte, macrophage or dendritic cell, the modified cell is recruited to the tumor microenvironment where it acts as a potent immune effector by infiltrating the tumor and killing the target cells. One aspect includes a modified cell and pharmaceutical compositions comprising the modified cell for adoptive cell therapy and treating a disease or condition associated with immunosuppression.
1. A pharmaceutical composition comprising:
(a) a population of modified human cells comprising a chimeric antigen receptor (CAR), wherein:
(i) at least 50% of the cells in the population of cells are CD14+ or CD11b+, and
(ii) the population of cells has been depleted for cells expressing CD3 and CD19 and
(b) a pharmaceutically acceptable excipient.
2. The pharmaceutical composition of claim 1 , wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule.
3. The pharmaceutical composition of claim 2 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain, an anti-mesothelin antigen binding domain, an anti-CD19 antigen binding domain or an anti-PSMA antigen binding domain.
4. The pharmaceutical composition of claim 2 , wherein the antigen binding domain of the CAR comprises an antibody selected from the group consisting of a monoclonal antibody, a polyclonal antibody, a synthetic antibody, human antibody, humanized antibody, single domain antibody, single chain variable fragment, and antigen-binding fragments thereof.
5. The pharmaceutical composition of claim 2 , wherein the transmembrane domain of the CAR comprises a CD8 or CD28 transmembrane domain.
6. The pharmaceutical composition of claim 2 , wherein the intracellular domain of the CAR comprises dual signaling domains.
7. The pharmaceutical composition of claim 2 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain.
8. The pharmaceutical composition of claim 1 , wherein at least 70% of the cells in the population of cells are CD14+ or CD11b+.
9. The pharmaceutical composition of claim 1 , wherein the modified human cells exhibit targeted effector activity.
10. The pharmaceutical composition of claim 9 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR.
11. The pharmaceutical composition of claim 9 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, targeted cellular cytotoxicity, antigen presentation, and cytokine secretion.
12. The pharmaceutical composition of claim 9 , wherein the targeted effector activity is enhanced by inhibition of CD47 or SIRPα activity.
13. The pharmaceutical composition of claim 1 , further comprising an agent selected from the group consisting of a nucleic acid, an antibiotic, an anti-inflammatory agent, an antibody or antibody fragments thereof, a growth factor, a cytokine, an enzyme, a protein, a peptide, a fusion protein, a synthetic molecule, an organic molecule, a carbohydrate or the like, a lipid, a hormone, a microsome, a derivative or a variation thereof, and any combination thereof.
14. The pharmaceutical composition of claim 1 , wherein the modified human cells exhibit reduced SIRPα activity relative to an unmodified human cell.
15. The pharmaceutical composition of claim 1 , wherein at least 35% of the cells in the composition express a CAR.
16. A method for stimulating an immune response to a target tumor cell or tumor tissue in a subject comprising administering to a subject a therapeutically effective amount of the pharmaceutical composition of claim 1 .
17. A method of modifying a population of human cells, comprising:
introducing a chimeric antigen receptor (CAR) into the population of human cells,
wherein after introducing the CAR into the population of human cells, at least 50% of the modified cells in the population of cells are CD14+ or CD11b+, and the population of cells has been depleted for cells expressing CD3 and CD19.
18. The method of claim 17 , wherein introducing the CAR into the population of human cells comprises introducing a nucleic acid sequence encoding the CAR.
19. The method of claim 18 , wherein introducing the nucleic acid sequence comprises electroporating an mRNA encoding the CAR into the population of human cells.
20. The method of claim 19 , wherein introducing the nucleic acid sequence comprises transducing the population of human cells with a viral vector comprising a nucleic acid sequence encoding the CAR.
21. The method of claim 17 , wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule.
22. The method of claim 21 , wherein the antigen binding domain comprises an anti-HER2 antigen binding domain, an anti-mesothelin antigen binding domain, an anti-CD-19 antigen binding domain, or an anti-PSMA antigen binding domain.
23. The method of claim 21 , wherein the transmembrane domain of the CAR comprises a CD8 or CD28 transmembrane domain.
24. The method of claim 21 , wherein the intracellular domain of the CAR comprises dual signaling domains.
25. The method of claim 21 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain.
26. The method of claim 21 , wherein at least 70% of the cells in the population of cells are CD14+ or CD11b+.
27. The method of claim 17 , wherein the modified human cells exhibit targeted effector activity.
28. The method of claim 27 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR.
29. The method of claim 27 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, targeted cellular cytotoxicity, antigen presentation, and cytokine secretion.
30. The method of claim 17 , wherein the modified human cells exhibit reduced SIRPα activity relative to an unmodified human cell.