Methods and compositions for treating cancer
A nucleic acid sequence is provided that encodes a chimeric protein comprising a ligand that comprises a naturally occurring or modified follicle stimulating hormone sequence, e.g., an FSHp sequence, or fragment thereof, which ligand binds to human follicle stimulating hormone (FSH) receptor, linked to either (a) a nucleic acid sequence that encodes an extracellular hinge domain, a transmembrane domain, a co-stimulatory signaling region, and a signaling endodomain; or (b) a nucleic acid sequence that encodes a ligand that binds to NKG2D. The vector containing the nucleic acid sequence, the chimeric proteins so encoded, and modified T cells expressing the chimeric protein, as well as method of using these compositions for the treatment of FSHR-expressing cancers or tumor cells are also provided.
1. A method of treating a cancer in a human subject comprising:
administering to a subject having a tumor that expresses follicle stimulating hormone receptor (FSHR), a modified human T cell that comprises a nucleic acid sequence that encodes a chimeric protein, wherein the chimeric protein comprises a ligand linked to an extracellular hinge domain, a transmembrane domain, a co-stimulatory signaling region, and a signaling endodomain,
wherein the ligand comprises an FSHβ subunit or a fragment thereof,
wherein the ligand is capable of binding to FSHR on the tumor that expresses FSHR, and wherein the chimeric protein is capable of activating the modified human T cell.
2. The method according to claim 1 , wherein the modified human T cell does not express Forkhead Box Protein (Foxp1).
3. The method according to claim 1 , wherein said modified human T cell is an autologous human T cell or natural killer (NK) T cell obtained from the subject or from a bone marrow transplant match for the subject.
4. The method according to claim 1 , wherein the tumor is ovarian cancer, cancer of the prostate, breast, colon, pancreas, urinary bladder, kidney, lung, liver, stomach, or testis, or the modified human T-cell binds to cells of blood vessels of either primary or metastatic tumors.
5. The method according to claim 1 , further comprising decreasing, ablating or down-regulating expression of Forkhead Box Protein (Foxp1) in said modified human T-cell.
6. The method according to claim 5 , wherein the manipulation occurs either
(a) before a T cell is transfected with the nucleic acid sequence that encodes the chimeric protein; or
(b) after a T cell is transfected with the nucleic acid sequence that encodes the chimeric protein.
7. The method according to claim 1 , further comprising, prior to administration,
(a) removing T cells from the subject;
(b) transducing the T cells ex vivo with a vector encoding the chimeric protein; and
(c) formulating the T cells in a suitable pharmaceutical carrier.
8. The method according to claim 1 , further comprising treating the subject with chemotherapy.
9. The method according to claim 1 , wherein said modified human T cell is an endogenous or heterologous human T cell or human T cell line.
10. The method according to claim 1 , wherein the co-stimulatory domain is selected from a CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, and B7-H3 costimulatory domain.
11. The method according to claim 1 , wherein the signaling endodomain is selected from a CD3ζ, TCR ζ, FcR γ, FcR β, CD3γ, CD3δ, CD3ε, CD5, CD22, 25 CD79a, CD79b, and CD66d signaling endodomain.
12. The method according to claim 1 , wherein the transmembrane domain is selected from a T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CDS, CD9, CD16, CD22, CD33, CD37, CD64, CDSO, CD86, CD134, CD137, and CD154 transmembrane domain.
13. The method according to claim 1 , wherein the ligand further comprises an FSHα subunit.
14. The method according to claim 1 , wherein the ligand comprises the FSHβ subunit linked to an FSHα subunit by a linker sequence.
15. The method according to claim 1 , wherein the FSHβ subunit is a first FSHβ subunit, wherein the ligand further comprises a second FSHβ subunit and a linker, and wherein the first FSHβ subunit is linked to the second FSHβ subunit by the linker sequence.
16. The method according to claim 1 , wherein the ligand comprises an FSHβ subunit fragment.
17. The method according to claim 16 , wherein the FSHβ subunit fragment comprises an amino acid sequence selected from the group consisting of amino acids 19-33 of SEQ ID NO: 2, 51-71 of SEQ ID NO:2, 69-83 of SEQ ID NO:2, and 99-113 of SEQ ID NO: 2.
18. The method according to claim 1 , wherein the ligand comprises a full length FSHβ subunit.