Methods and compositions for enhancing the therapeutic effect of anti-tumor T cells
Compositions, e.g., therapeutic agents, and methods are provided for modulating gene and protein expression of Forkhead Box protein 1 (Foxp1). The therapeutic agents include short nucleic acid molecules that modulate gene and protein expression of Forkhead Box protein 1 (Foxp1) expression, viral vectors containing such molecules, T cells transduced with these viruses for adoptive therapies, and any small molecules that bind to and inactivate Foxp1. These compounds and methods have applications in cancer therapy either alone or in combination with other therapies that stimulate the endogenous immune system in the environment of the cancer, e.g., tumor.
1. A method of preparing a composition for treating a cancer comprising:
(a) pulsing T cells with an ovarian cancer antigen; and
(b) transducing said pulsed T cells with a first vector expressing a construct that down regulates or extinguishes Foxp1, said first vector comprising the short hairpin (shRNA) sequence SEQ ID NO: 5 downstream of a selected promoter; and
(c) formulating said pulsed, transfected T cells with a suitable pharmaceutical carrier.
2. A method of preparing a composition for treating ovarian cancer comprising:
(a) pulsing T cells with an ovarian cancer antigen; and
(b) transducing said pulsed T cells with a first vector that targets a T cell receptor or T cell surface marker or co-expresses a TCR or chimeric anti-tumor receptor, and
(c) treating said pulsed, transfected T cells with a zinc-finger nuclease designed to target a unique sequence of Foxp1;
(d) culturing the T cells of step (c) to remove Foxp1 from the cells ex vivo; and
(e) formulating the T cells in a suitable pharmaceutical carrier.
3. A method of preparing a composition for treating a cancer comprising:
transducing or transfecting a T cell ex vivo with a first vector expressing a construct that inhibits, decreases or extinguishes the expression of Foxp1, said vector comprising the short hairpin (shRNA) sequence SEQ ID NO: 5 downstream of a selected promoter;
transfecting the T cell ex vivo with a second vector that targets a T cell receptor or T cell surface marker or co-expresses an anti-tumor T cell receptor or a chimeric anti-tumor T cell receptor, and
formulating the T cells in a pharmaceutically acceptable carrier or diluent that is suitable for administration to a human subject.
4. The method according to claim 3 , wherein the first vector and second vector are the same vector.
5. The method according to claim 3 , wherein the T cell is
(a) a polyclonal or monoclonal tumor-reactive T cell;
(b) a tumor-infiltrating lymphocyte generated from aphaeresis samples or isolated from a tumor or peripheral blood of a cancer patient; or
(c) a T cell that has been conditioned for adoptive transfer by pulsing ex vivo with a tumor-specific antigen before it is transduced with the virus vector.
6. The method according to claim 1 , wherein the first vector is a lentiviral vector and the promoter is an RNA polymerase III promoter.
7. The method according to claim 2 , wherein the first vector is a lentiviral vector and the promoter is an RNA polymerase III promoter.
8. The method according to claim 3 , wherein the first and second vector is a lentiviral vector and the promoter for each vector is an RNA polymerase III promoter.
9. The method according to claim 2 , wherein the T cell surface marker is CD3, CTLA4, CD44, CD69 or CD25.
10. The method according to claim 3 , wherein the T cell surface marker is CD3, CTLA4, CD44, CD69 or CD25.
11. The method according to claim 2 , wherein the anti-tumor T cell receptor targets human carcino embryonic antigen, NY-ESO-1, MAGE-A3, MART-1, Gp100 or p53.
12. The method according to claim 3 , wherein the anti-tumor T cell receptor targets human carcino embryonic antigen, NY-ESO-1, MAGE-A3, MART-1, Gp100 or p53.
13. The method according to claim 2 , wherein the chimeric anti-tumor T cell receptor targets CD19, Epstein Barr virus, mesothelin or folate receptor.
14. The method according to claim 3 , wherein the chimeric anti-tumor T cell receptor targets CD19, Epstein Barr virus, mesothelin or folate receptor.