RNA ENGINEERED T CELLS FOR THE TREATMENT OF CANCER
The present invention relates to compositions and methods for generating RNA Chimeric Antigen Receptor (CAR) transfected T cells. The RNA-engineered T cells can be used in adoptive therapy to treat cancer.
1 .- 30 . (canceled)
31 . A method of generating a population of RNA-engineered T cells transiently expressing exogenous RNA, the method comprising introducing an in vitro transcribed RNA or synthetic RNA into a T cell, wherein the RNA comprises a nucleic acid sequence encoding an extracellular domain, a transmembrane domain, a costimulatory signaling region, and a signaling domain of CD3-zeta.
32 .- 45 . (canceled)
46 . A method of treating a cancer patient, the method comprising administering to the patient a T cell engineered to transiently express exogenous RNA, wherein the RNA comprises a nucleic acid sequence encoding an extracellular domain, a transmembrane domain, a costimulatory signaling region, and a signaling domain of CD3-zeta.
47 . The method of claim 46 , wherein the extracellular domain comprises an antigen binding moiety.
48 . The method of claim 47 , wherein the antigen bind moiety binds to a tumor antigen.
49 . The method of claim 48 , wherein the tumor antigen is an antigen associated with a cancer 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 any combination thereof.
50 . The method of claim 46 comprising repeating the administration of a T cell.
51 . The method of claim 46 comprising administering a chemotherapeutic agent to the patient.
52 . The method of claim 46 , wherein the RNA is transcribed from an in vitro transcription vector, wherein the vector is pD-A.ss1.OF.BBZ.2bg.150A, wherein the vector comprises the nucleic acid sequence of SEQ ID NO: 4.
53 . The method of claim 46 , wherein the DNA from which the RNA is transcribed comprises a sequence selected from the group consisting of SEQ ID NO: 6 and SEQ ID NO: 8.
54 . The method of claim 46 , wherein the RNA is transcribed from an in vitro transcription vector wherein the vector is pD-A.19.OF.2bg.150A, wherein the vector comprises the nucleic acid sequence of SEQ ID NO: 5.
55 . The method of claim 46 , wherein the DNA from which the RNA is transcribed comprises a sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24.
56 . The method of claim 46 , wherein the RNA is transcribed from an in vitro transcription vector, wherein the vector is pD-A.GD2.OF.8TMBBZ.2bg.150A, wherein the vector comprises the nucleic acid sequence of SEQ ID NO: 28.
57 . The method of claim 46 , wherein the DNA from which the RNA is transcribed comprises a sequence selected from the group consisting of SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13.
58 . The method of claim 46 , wherein the RNA is transcribed from an in vitro transcription vector, wherein the vector is pD-A.cMet.OF.8TMBBZ.2bgUTR.150A, wherein the vector comprises the nucleic acid sequence of SEQ ID NO: 27.
59 . The method of claim 46 , wherein the DNA from which the RNA is transcribed 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 and SEQ ID NO: 18.
60 . The method of claim 46 , wherein the costimulatory signaling region comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof.
61 . The method of claim 46 , wherein the nucleic acid sequence comprises a poly(A) tail comprising 150 adenosine bases.
62 . The method of claim 46 , wherein the nucleic acid sequence comprises a 3′UTR comprising at least one repeat of a 3′UTR derived from human beta-globulin.