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 modified macrophage or monocyte 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, and
wherein the modified macrophage or monocyte exhibits reduced SIRPα activity relative to an unmodified cell.
2. The modified macrophage or monocyte of claim 1 , wherein the modified macrophage or monocyte comprises deletion of SIRPα using CRISPR/Cas9.
3. The modified macrophage or monocyte of claim 1 , wherein the modified macrophage or monocyte has been treated with an anti-SIRPα antibody.
4. The modified macrophage or monocyte of claim 1 , wherein the modified macrophage or monocyte comprises anti-SIRPα antibodies.
5. The modified macrophage or monocyte of claim 1 , wherein the modified macrophage or monocyte comprises anti-SIRPα siRNA.
6. The modified macrophage or monocyte of claim 1 , wherein the modified macrophage or monocyte exhibits increased targeted effector activity relative to an unmodified cell.
7. The modified macrophage or monocyte of claim 6 , wherein the increased targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR.
8. The modified macrophage or monocyte of claim 6 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, targeted cellular cytotoxicity, antigen presentation, and cytokine secretion.
9. The modified macrophage or monocyte 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.
10. The modified macrophage or monocyte of claim 1 , wherein the antigen binding domain of the CAR is selected from the group consisting of an anti-CD19 antibody, an anti-HER2 antibody, and a fragment thereof.
11. The modified macrophage or monocyte of claim 1 , wherein the transmembrane domain of the CAR comprises a CD8 or CD28 transmembrane domain.
12. The modified macrophage or monocyte of claim 1 , wherein the intracellular domain of the CAR comprises dual signaling domains.
13. The modified macrophage or monocyte of claim 1 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain.
14. The modified macrophage or monocyte 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.
15. A pharmaceutical composition comprising the modified macrophage or monocyte of claim 1 and a pharmaceutically acceptable carrier.
16. A method of treating a disease or condition associated with a tumor or cancer in a subject comprising:
administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising the modified macrophage or monocyte of claim 1 .
17. A method of modifying a macrophage or monocyte, comprising:
introducing a nucleic acid encoding a chimeric antigen receptor (CAR) into the macrophage or monocyte,
wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule, and
introducing CRISPR/Cas9 gene editing material into the macrophage or monocyte,
wherein the modified macrophage or monocytes exhibits targeted effector activity and reduced SIRPα activity.
18. A method of modifying a macrophage or monocyte, comprising:
introducing a nucleic acid encoding a chimeric antigen receptor (CAR) into the macrophage or monocyte,
wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule, and
wherein the macrophage or monocyte is engineered to encode an siRNA that downregulates SIRPα,
wherein the modified macrophage or monocytes exhibits targeted effector activity and reduced SIRPα activity.
19. A method of modifying a macrophage or monocyte, comprising:
introducing a nucleic acid encoding a chimeric antigen receptor (CAR) into the macrophage or monocyte,
wherein the CAR comprises an antigen binding domain, a transmembrane domain and an intracellular domain of a stimulatory and/or co-stimulatory molecule, and
wherein the macrophage or monocyte is capable of secreting an anti-SIRPα antibody,
wherein the modified macrophage or monocytes exhibits targeted effector activity and reduced SIRPα activity.
20. The method of claim 17 , wherein introducing the CAR into the macrophage or monocyte comprises introducing a nucleic acid sequence encoding the CAR.
21. The method of claim 17 , wherein the targeted effector activity is directed against a target cell comprising an antigen that specifically binds the antigen binding domain of the CAR.
22. The method of claim 17 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, targeted cellular cytotoxicity, antigen presentation, and cytokine secretion.
23. The method of claim 17 , 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.
24. The method of claim 17 , wherein the antigen binding domain of the CAR is selected from the group consisting of an anti-CD19 antibody, an anti-HER2 antibody, and a fragment thereof.