Chimeric antigen receptor dendritic cell (CAR-DC) for treatment of cancer
The current invention provides monocytic cells transfected with chimeric antigen receptor (CAR) to selectively home to tumors and upon homing differentiate into dendritic cells capable of activating immunity which is inhibitory to said tumor. In one embodiment of the invention, monocytic cells are transfected with a construct encoding an antigen binding domain, a transcellular or structural domain, and an intracellular signaling domain. In one specific aspect of the invention, the antigen binding domain interacts with sufficient affinity to a tumor antigen, capable of triggering said intracellular domain to induce an activation signal to induce monocyte differentiation into DC.
1. A method of treating a HER2/neu expressing cancer in a human subject in need thereof comprising administering to the human subject a pharmaceutical composition comprising
(a) a therapeutically effective amount of an ex vivo population of phagocytic CD14+/CD16+cells comprising a recombinant polynucleic acid with a sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises (i) an extracellular domain comprising an anti-HER2/neu binding domain and a CD8a hinge domain; (ii) a transmembrane domain; and (iii) an intracellular domain containing a CD3 zeta intracellular signaling domain; and
(b) a pharmaceutically acceptable carrier or excipient;
thereby treating the cancer in the human subject.
2. The method of claim 1 , wherein the intracellular domain comprises two or more intracellular signaling domains.
3. The method of claim 1 , wherein the transmembrane domain comprises a CD8a transmembrane.
4. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells is from the human subject.
5. The method of claim 4 , wherein the ex vivo population of phagocytic CD14+/CD16+cells is from a leukapheresis sample, a blood sample, or a PBMC sample from the human subject.
6. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells is an ex vivo population of virally transduced cells.
7. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells comprises a viral component.
8. The method of claim 1 , wherein the anti-HER2/neu binding domain is a single chain variable fragment (scFv).
9. The method of claim 1 , wherein the sequence of the recombinant polynucleic acid encoding the CAR is from a viral vector.
10. The method of claim 9 , wherein the method further comprises transducing the viral vector into a population of CD14+cells ex vivo, thereby obtaining the ex vivo population of phagocytic CD14+/CD16+cells comprising the recombinant polynucleic acid with a sequence encoding a CAR.
11. The method of claim 1 , wherein the method comprises (i) extracting a blood sample from the human subject; (ii) isolating CD14+cells from the blood sample; and (iii) transfecting the CD14+cells from (ii) with the recombinant polynucleic acid with a sequence encoding a CAR; and wherein administering comprises infusing.
12. The method of claim 1 , wherein the recombinant polynucleic acid is mRNA.
13. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells stimulates killing of cancer cells in the human subject by T cells of the human subject.
14. The method of claim 1 , wherein the intracellular domain of the CAR is capable of inducing monocytic differentiation to M1 macrophages in the human subject.
15. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells enhances or improves effector function of a T cell in the human subject.
16. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells directly kills cancer cells in the human subject.
17. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells inhibits macrophage or macrophage related cells of the human subject from promoting tumor growth.
18. The method of claim 1 , wherein the ex vivo population of phagocytic CD14+/CD16+cells is an ex vivo population of isolated CD14+/CD16+cells.
19. The method of claim 1 , wherein the cancer is a lymphoma.
20. The method of claim 1 , wherein the cancer is a solid tumor.
21. The method of claim 20 , wherein the cancer is a breast cancer.
22. The method of claim 20 , wherein the cancer is a metastatic cancer.
23. The method of claim 1 , wherein the method further comprises administering GM-CSF, IL-2, an agent that blocks CD47 activity or an agent that induces immunogenic cell death to the human subject.