Compositions and methods for T cell delivery of therapeutic molecules
The present invention includes compositions and methods for modifying a T cell with a nucleic acid encoding a switch molecule comprising an extracellular domain comprising a membrane receptor or fragment thereof and an intracellular domain comprising a signaling receptor or fragment thereof. In one aspect, a method comprises introducing a nucleic acid encoding a switch molecule and a nucleic acid encoding a soluble fusion protein and/or a nucleic acid encoding a bispecific antibody into a population of cells comprising T cells, wherein the T cells transiently expresses the switch molecule and soluble fusion protein or bispecific antibody. In other aspect, compositions of T cells and methods of treating a disease or condition, such as cancer or an autoimmune disease, are also included.
1. A modified T cell comprising a nucleic acid encoding a switch molecule, wherein the switch molecule comprises: an extracellular domain of a transforming growth factor-beta receptor (TGF-beta-R); and an intracellular domain of a signaling receptor, wherein the signaling receptor is selected from the group consisting of interleukin-12 receptor (IL-12R), CD3, CD28, CD137, CD27, ICOS, OX40, and any combination thereof, wherein the T cell expresses the switch molecule and interaction of the extracellular domain of the switch molecule with TGF-beta induces the T cell to secrete an activation factor at a target site.
2. The modified T cell of claim 1 , wherein the signaling receptor is interleukin-12 receptor (IL-12R).
3. The modified T cell of claim 1 , wherein the activation factor is a soluble cytokine selected from the group consisting of IL-1, IL-2, IL-6, IL-7, IL-8, IL-10, IL-12, IL-15, IL-21, TNF, TGF, IFN, and functional fragments and variants thereof.
4. The modified T cell of claim 1 , wherein the nucleic acid comprises in vitro transcribed RNA or synthetic RNA.
5. The modified T cell of claim 1 , wherein the target site is a tumor selected from the group consisting of brain cancer, bladder cancer, breast cancer, cervical cancer, colorectal cancer, liver cancer, kidney cancer, lymphoma, leukemia, lung cancer, melanoma, metastatic melanoma, mesothelioma, neuroblastoma, ovarian cancer, prostate cancer, pancreatic cancer, renal cancer, skin cancer, thymoma, sarcoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, uterine cancer, and combinations thereof.
6. The modified T cell of claim 1 , wherein the T cell homes to a solid tumor site.
7. The modified T cell of claim 1 , wherein the T cell homes to a tumor antigen.
8. The modified T cell of claim 7 , wherein the tumor antigen is selected from the group consisting of p53, Ras, beta-Catenin, CDK4, alpha-Actinin-4, Tyrosinase, TRP1/gp75, TRP2, gp100, Melan-A/MART1, Gangliosides, PSMA, HER2, WT1, EphA3, EGFR, CD20, MAGE, BAGE, GAGE, NY-ESO-1, Telomerase, Survivin, and any combination thereof.
9. The modified T cell of claim 1 , wherein the T cell is activated.
10. A composition comprising the modified T cell of claim 1 and further comprising a pharmaceutically acceptable carrier.
11. The modified T cell of claim 1 , further comprising a nucleic acid that encodes a chimeric antigen receptor (CAR).
12. The modified T cell of claim 11 , wherein the intracellular domain of the switch molecule comprises the intracellular domain of the interleukin-12 receptor (IL-12R).
13. A method for generating a T cell expressing a switch molecule comprising introducing a nucleic acid encoding a switch molecule into a population of T cells, wherein the switch molecule comprises: an extracellular domain of a transforming growth factor-beta receptor (TGF-beta-R); and an intracellular domain of a signaling receptor, wherein the signaling receptor is selected from the group consisting of interleukin-12 receptor (IL-12R), CD3, CD28, CD137, CD27, ICOS, OX40, and any combination thereof, and
wherein the T cells express the switch molecule and interaction of the extracellular domain of the switch molecule with TGF-beta induces the T cells to secrete an activation factor at a target site, thereby generating the T cell.
14. The method of claim 13 , further comprising activating the T cells to home to the target site, wherein the T cells secrete the activation factor at the target site.
15. The method of claim 13 , wherein the population of T cells is within cells selected from the group consisting of peripheral blood mononuclear cells, cord blood cells, a purified population of T cells, and a T cell line.
16. The method of claim 15 , wherein the population of T cells is within peripheral blood mononuclear cells.
17. The method of claim 15 , wherein the population of T cells is within purified T cells.
18. The method of claim 13 , wherein the population of T cells are cryopreserved.
19. The method of claim 18 further comprising thawing the cryopreserved T cells.
20. The method of claim 13 , wherein the nucleic acid comprises in vitro transcribed RNA or synthetic RNA.
21. The method of claim 13 , further comprising introducing a nucleic acid that encodes a chimeric antigen receptor (CAR).
22. The method of claim 21 , wherein the intracellular domain of the switch molecule comprises the intracellular domain of the interleukin-12 receptor (IL-12R).
23. A method for generating a T cell transiently expressing a switch molecule and bispecific antibody comprising:
introducing a nucleic acid encoding a switch molecule comprising an extracellular domain comprising a membrane receptor or fragment thereof and an intracellular domain comprising a signaling receptor or fragment thereof and at least one of a nucleic acid encoding a soluble fusion protein and a nucleic acid encoding a bispecific antibody comprising bispecificity for an antigen on a target cell and an antigen on an activating T cell, and wherein the T cells transiently express the switch molecule and soluble fusion protein and/or bispecific antibody and activation of the T cells induces secretion of an activation factor at a target site.