Compositions and methods of cellular immunotherapy
Disclosed herein are methods of treating a subject exhibiting a solid tumor that expresses Glypican-3 (GPC3). The methods typically utilize g GPC3 chimeric antigen receptor immunoresponsive cells to a subject in need thereof to effect killing of tumor cells.
1. A method of treating a subject exhibiting a solid tumor that expresses Glypican-3 (GPC3), the method comprising administering anti-GPC3 chimeric antigen receptor immunoresponsive cells to the subject, wherein the administering takes place after or concurrent with subjecting the subject to a lymphocyte reduction treatment, and wherein the lymphocyte reduction treatment comprises administering cyclophosphamide and fludarabine to the subject.
2. The method of claim 1 , wherein the immunoresponsive cells are NK cells (anti-GPC3-CAR NK cells) or T cells (anti-GPC3-CAR T cells).
3. The method of claim 2 , wherein the administering the anti-GPC3-CAR T cells to the subject takes place after subjecting the subject to the lymphocyte reduction treatment.
4. The method of claim 2 , wherein at least about 5×10 4 anti-GPC3-CAR T cells/kg are administered to the subject.
5. The method of claim 2 , wherein from about 5×10 4 to about 1×10 12 anti-GPC3-CAR T cells/kg are administered to the subject.
6. The method of claim 1 , wherein the administration is effective in reducing tumor size by at least 30% as measured by computerized tomography (CT) scan.
7. The method of claim 1 , wherein the administration is effective in stabilizing tumor size as measured by a less than 10% change in a baseline measurement of a diameter of the tumor lesion as measured by computerized tomography (CT) scan.
8. The method of claim 2 , wherein the administering of anti-GPC3-CAR T cells and the subjecting the subject to the lymphocyte reduction treatment synergistically increase the subject's medium survival time by at least about 6 months as compared to administering the anti-GPC3-CAR T cells alone.
9. The method of claim 1 , wherein the solid tumor is liver cancer, stomach cancer, lung cancer, breast cancer, head and neck cancer, ovarian cancer, thyroid cancer, kidney cancer, bladder cancer, cervical cancer, pancreatic cancer, liposarcoma, testicular nonseminomatous germ cell cancer, melanoma, adenoma of the adrenal gland, schwannoma, malignant fibrous histiocytoma, or esophageal cancer.
10. The method of claim 1 , wherein the anti-GPC3 chimeric antigen receptor comprises an antigen binding unit that exhibits specific binding to C-terminus of GPC3.
11. The method of claim 10 , wherein the antigen binding unit comprises a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8.
12. The method of claim 1 , wherein the anti-GPC3-CAR comprises a sequence selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, or SEQ ID NO: 30.
13. The method of claim 1 , wherein the anti-GPC3-CAR comprises a sequence selected from the group consisting of SEQ ID NO 28, SEQ ID NO:29 or SEQ ID NO 30.
14. The method of claim 2 , wherein the anti-GPC3-CAR T cells comprise at least two intracellular signaling domains.
15. The method of claim 2 , wherein the anti-GPC3-CAR T cells comprise at least three intracellular signaling domains.
16. The method of claim 14 , wherein the intracellular signaling domains are selected from a signaling domain derived from CD3, CD28, 4-1BB, OX40, DAP10, or ICOS.
17. The method of claim 1 , wherein the lymphocyte reduction treatment comprises reducing a quantity of regulatory T cells in the subject.
18. The method of claim 17 , wherein the reducing the quantity of regulatory T cells comprises a reduction of at least about 30% of the regulatory T cells as measured by flow cytometric analysis of circulating CD4 + and CD25 + cells in the subject.
19. The method of claim 1 , wherein the lymphocyte reduction treatment further comprises administering radiation or a biological agent to the subject.
20. The method of claim 1 , wherein the lymphocyte reduction treatment further comprises administering another chemotherapy to the subject.
21. The method of claim 20 , wherein the administering another chemotherapy to the subject comprises administering a chemotherapeutic agent selected from the group consisting of etoposide, cytarabine, methotrexate, vincristine adriamycin, and any combination thereof.
22. The method of claim 21 , wherein the chemotherapeutic agent is administered to the subject at least one time prior to administration of the anti-GPC3-CAR T cells.
23. The method of claim 1 , wherein the lymphocyte reduction treatment reduces a quantity of lymphocytes by at least about 20% as measured by complete blood count (CBC) analysis.
24. The method of claim 2 , further comprising a second administration of the anti-GPC3-CAR T cells to the subject.
25. The method of claim 1 , wherein the subject has refractory, persistent, or progressive disease.
26. The method of claim 2 , wherein the anti-GPC3-CAR T cells are autologous or allogenic to the subject.
27. The method of claim 1 , further comprising administering at least one immunostimulatory agent to the subject concurrent or after administration of the anti-GPC3 chimeric antigen receptor immunoresponsive cells.
28. The method of claim 27 , wherein the immunostimulatory agent is selected from the group consisting of aldesleukin (IL-2), IL-3, IL-6, IL-11, GM-CSF, and any combination thereof.
29. The method of claim 19 , wherein the biological agent is an antibody directed to an antigen expressed on a lymphocyte.