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 population of modified human monocytes comprising a chimeric antigen receptor (CAR),
wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule,
wherein at least 50% of the modified human monocytes in the population of cells express the CAR, and
wherein at least 70% of the modified human monocytes are viable.
2. The population of claim 1 , wherein the antigen binding domain comprises an anti-HER2, anti-mesothelin, anti-CD19, or anti-PSMA antigen binding domain.
3. The population of claim 1 , 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.
4. The population of claim 1 , wherein the transmembrane domain of the CAR comprises a CD8 or CD28 transmembrane domain.
5. The population of claim 1 , wherein the intracellular domain of the CAR comprises dual signaling domains.
6. The population of claim 1 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain.
7. The population of claim 1 , wherein the modified human monocytes exhibit targeted effector activity.
8. The population of claim 7 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR.
9. The population of claim 7 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, targeted cellular cytotoxicity, antigen presentation, and cytokine secretion.
10. The population of claim 7 , wherein the targeted effector activity is enhanced by inhibition of CD47 or SIRPα activity.
11. The population 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.
12. The population of claim 1 , wherein the modified human monocytes exhibit reduced SIRPα activity relative to unmodified human monocytes.
13. A pharmaceutical composition comprising the population of modified human monocytes of claim 1 and a pharmaceutically acceptable carrier.
14. A method for stimulating an immune response to a target tumor cell or tumor tissue in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising the modified human monocytes of claim 1 .
15. A method of modifying a population of human monocytes, comprising:
introducing a chimeric antigen receptor (CAR) into the population of human monocytes,
wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule,
wherein after introducing the CAR into the population of human monocytes, at least 50% of the monocytes in the population of modified human monocytes express the CAR and at least 70% of the modified human monocytes are viable.
16. The method of claim 15 , wherein introducing the CAR into the population of human monocytes comprises introducing a nucleic acid sequence encoding the CAR.
17. The method of claim 16 , wherein introducing the nucleic acid sequence comprises electroporating an mRNA encoding the CAR.
18. The method of claim 17 , wherein introducing the nucleic acid sequence comprises transducing the cell with a viral vector comprising a nucleic acid sequence encoding the CAR.
19. The method of claim 16 , wherein the modified human monocytes exhibit targeted effector activity.
20. The method of claim 19 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, targeted cellular cytotoxicity, antigen presentation, and cytokine secretion.
21. The method of claim 15 , further comprising inhibiting CD47 or SIRPα activity to enhance the targeted effector activity.
22. The method of claim 21 , wherein inhibiting CD47 or SIRPα activity comprises contacting the human monocyte with a blocking anti-CD47 antibody or a blocking anti-SIRPα antibody.
23. The method of claim 15 , wherein the antigen binding domain comprises an anti-HER2, anti-mesothelin, anti-CD19, or anti-PSMA antigen binding domain.
24. The method of claim 15 , wherein the antigen binding domain of the CAR comprises an antibody selected from the group consisting of a synthetic antibody, human antibody, humanized antibody, single domain antibody, single chain variable fragment, and antigen-binding fragments thereof.