T-Rapa cells as novel effector cell type for chimeric antigen receptor therapy
The present disclosure provides chimeric antigen receptor (CAR)-T-Rapa cells and methods of making and using them. Specifically, methods of producing T-Rapa cells that can express chimeric antigen receptors is provided.
1. A method of making a T- Rapa cell expressing a chimeric antigen receptor (CAR) comprising the steps of:
(a) conditioning ex vivo CD3+T-cells with rapamycin to generate T-cells resistant to rapamycin (T- Rapa cells); and
(b) transducing the T- Rapa cells in vitro with a vector that expresses the CAR to produce CAR-T- Rapa cells.
2. A CAR-T- Rapa cell comprising:
an exogenous vector encoding and capable of expressing a chimeric antigen receptor (CAR), wherein the T- Rapa cells are made by culturing ex vivo CD3+T-cells in chemically defined medium comprising rapamycin for sufficient time to generate T-cells resistant to rapamycin (T- Rapa cells).
3. The CAR-T- Rapa cell of claim 2 , wherein the T cells are obtained from a subject or suitable donor.
4. The CAR T- Rapa cell of claim 2 , wherein the vector is a viral vector.
5. The CAR T- Rapa cell of claim 2 , wherein the T cells are obtained from the subject or suitable donor by
(i) obtaining a sample from the subject; and
(ii) detecting and isolating CD3+T-cells from the sample and culturing the isolated T-cells in vitro.
6. The CAR T- Rapa cell of claim 2 , wherein the vector is a lentiviral vector.
7. The CAR T- Rapa cell of claim 2 , wherein the chemically defined medium comprises about 0.1 to about 2 micromolar rampamycin.
8. The CAR T- Rapa cells of claim 7 , wherein the chemically defined medium comprises IL-2 and IFN-α.
9. The CAR T- Rapa cell of claim 2 , wherein the CAR is a CAR specific to a tumor antigen.
10. The CAR T- Rapa cell of claim 2 , wherein the CAR is a CD19 CAR.
11. A method of treating a subject diagnosed with a disease that is treatable with CAR therapy, the method comprising:
administering an effective amount of the CAR-T- Rapa cell of claim 2 to treat the subject.
12. The method of claim 11 , wherein the disease is cancer.
13. The method of claim 12 , wherein the cancer is a hematologic cancer.
14. The method of claim 12 , wherein the cancer is a solid tumor.
15. The method of claim 11 , wherein the CAR-T- Rapa cells are introduced by intravenous transfusion.
16. A population comprising the CAR T- Rapa cells of claim 2 , wherein at least 95% of the T Rapa cells express one or more biomarkers indicative of CAR expression.
17. The population of claim 16 , wherein the population is capable of maintaining and expanding in_in vitro culture.
18. A stored population of CAR T Rapa cells, comprising the population of claim 17 , a storage solution, and wherein the population is stored at or below about −80° C. in suitable storage solution until used.
19. A CAR-T- Rapa cell comprising:
an exogenous vector encoding and capable of expressing a chimeric antigen receptor (CAR), wherein the CAR-T- Rapa cell is resistant to rapamycin.