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 pharmaceutical composition comprising:
(a) an ex vivo population of cells comprising CD14+/CD16+ cells, wherein the CD14+/CD16+ cells comprise a recombinant polynucleic acid with a sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises (i) 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 excipient;
wherein the pharmaceutical composition stimulates tumor killing by T cells in a subject with cancer when the pharmaceutical composition is administered to the subject.
2. The pharmaceutical composition of claim 1 , wherein the transmembrane domain comprises a CD8a transmembrane domain.
3. The pharmaceutical composition of claim 1 , wherein the intracellular domain comprises two or more intracellular signaling domains.
4. The pharmaceutical composition of claim 1 , wherein the ex vivo population of cells is an ex vivo population of virally transduced cells.
5. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells comprise a viral component.
6. The pharmaceutical composition of claim 1 , wherein the sequence of the recombinant polynucleic acid encoding the CAR is from a viral vector.
7. The pharmaceutical composition of claim 1 , wherein the ex vivo population of cells consists of CD14+/CD16+ cells.
8. The pharmaceutical composition of claim 1 , wherein the ex vivo population of cells is from a human subject.
9. The pharmaceutical composition of claim 8 , wherein the ex vivo population of cells is from a leukapheresis sample, a blood sample, or a PBMC sample from the human subject.
10. The pharmaceutical composition of claim 8 , wherein the human subject has cancer.
11. The pharmaceutical composition of claim 10 , wherein the cancer is a lymphoma or a solid tumor.
12. The pharmaceutical composition of claim 11 , wherein the cancer is breast cancer or metastatic cancer.
13. The pharmaceutical composition of claim 10 , wherein the cancer is an ErbB-2-expressing cancer.
14. The pharmaceutical composition of claim 1 , wherein the anti-HER2/neu binding domain is a single chain variable fragment (scFv).
15. The pharmaceutical composition of claim 1 , wherein the anti-HER2/neu binding domain is a single domain antibody (sdAb).
16. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells are phagocytic CD14+/CD16+ cells.
17. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells are human CD14+/CD16+ cells.
18. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells are isolated CD14+/CD16+ cells.
19. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells are monocytes.
20. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells are dendritic cells.
21. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells are macrophages.
22. The pharmaceutical composition of claim 1 , wherein the recombinant polynucleic acid is RNA.
23. The pharmaceutical composition of claim 22 , wherein the recombinant polynucleic acid is mRNA.
24. The pharmaceutical composition of claim 1 , wherein the recombinant polynucleic acid is from a viral vector.
25. The pharmaceutical composition of claim 1 , wherein the CAR consists of an anti-HER2/neu binding domain, a CD8a hinge domain, a CD8a transmembrane domain,
and an intracellular domain containing a CD3 zeta intracellular signaling domain.
26. The pharmaceutical composition of claim 1 , wherein the ex vivo population of cells comprises at least 1×10{circumflex over ( )} 6 CD14+/CD16+ cells.
27. The pharmaceutical composition of claim 1 , wherein the intracellular domain of the CAR is capable of inducing monocytic differentiation to M1 macrophages when the pharmaceutical composition is administered to a subject.
28. The pharmaceutical composition of claim 1 , wherein the CD14+/CD16+ cells of the pharmaceutical composition directly kill tumor cells when the pharmaceutical composition is administered to a subject with a tumor.
29. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition enhances or improves effector function of a T cell in a subject when the pharmaceutical composition is administered to the subject.
30. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition inhibits macrophage or macrophage related cells from promoting tumor growth when the pharmaceutical composition is administered to a subject with cancer.