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 cell 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 cell expresses the CAR and possesses targeted effector activity,
wherein the modified cell comprises a macrophage or a monocyte, and
wherein the modified cell comprises a lentiviral component.
2. The modified cell of claim 1 , wherein the modified cell exhibits reduced SIRPα activity relative to an unmodified monocyte or macrophage.
3. The modified cell 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 modified cell 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, an anti-mesothelin antibody, an anti-PSMA antibody, and a fragment thereof.
5. The modified cell of claim 1 , wherein the transmembrane domain of the CAR comprises a CD8 or CD28 transmembrane domain.
6. The modified cell of claim 1 , wherein the intracellular domain of the CAR comprises dual signaling domains.
7. The modified cell of claim 1 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain.
8. The modified cell of claim 1 , 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 modified cell of claim 1 , wherein the targeted effector activity is selected from the group consisting of phagocytosis, targeted cellular cytotoxicity, antigen presentation, and cytokine secretion.
10. The modified cell 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.
11. The modified cell of claim 1 , wherein the modified cell has at least one upregulated M1 marker and at least one downregulated M2 marker.
12. The modified cell of claim 1 , wherein the modified cell is genetically modified to express the CAR.
13. The modified cell of claim 1 , wherein the targeted effector activity is enhanced by inhibition of CD47 or SIRPα activity.
14. A pharmaceutical composition comprising the cell of claim 1 and a pharmaceutically acceptable carrier.
15. The pharmaceutical composition of claim 14 , 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 a pharmaceutical composition comprising the modified cell of claim 1 .
17. A method of modifying a cell, comprising:
introducing a chimeric antigen receptor (CAR) into the cell, 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 cell expresses the CAR and possesses targeted effector activity,
wherein the modified cell comprises a macrophage or a monocyte, and
wherein the modified cell comprises a lentiviral component.
18. The method of claim 17 , wherein introducing the CAR into the cell comprises introducing a nucleic acid sequence encoding the CAR.
19. The method of claim 17 , wherein introducing the nucleic acid sequence comprises electroporating an mRNA encoding the CAR.
20. 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.
21. 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.
22. The method of claim 17 , further comprising inhibiting CD47 or SIRPα activity to enhance the targeted effector activity.
23. The method of claim 22 , wherein inhibiting CD47 or SIRPα activity comprises contacting the cell with a blocking anti-CD47 antibody or a blocking anti-SIRPα antibody.
24. The method of claim 17 , wherein the intracellular domain of the CAR comprises dual signaling domains.
25. The method of claim 17 , wherein the intracellular domain of the CAR comprises a CD3 zeta intracellular domain.
26. The method of claim 17 , 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.
27. 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, an anti-mesothelin antibody, an anti-PSMA antibody, and a fragment thereof.