Allogeneic tumor cell vaccine

Abstract

The described invention provides allogeneic tumor cell vaccines comprising tumor cell lines or tumor cell line variants that are genetically engineered to express a core group of three immunomodulatory molecules, and optionally additional R immunomodulatory polypeptides for induction of one or more subpopulations of PBMCs to proliferate in response to the expressed immunomodulatory molecules and to then enter an effector phase for killing of tumor cells. According to some embodiments, the tumor cell vaccine candidate can induce an immune response in the recipient cancer patient that cross reacts with the patient's own (autologous) tumor cells, the effects of which are sufficient to result in enhanced anti-tumor immunity contributing to the increased survival of a vaccinated patient cohort compared to a matched unvaccinated patient cohort.

1 . An allogeneic tumor cell vaccine comprising:
 (1) a population of live, proliferation-incompetent genetically engineered tumor cells expressing one or more tumor specific antigens, the population comprising at least three stably expressed immunomodulatory molecules, wherein the at least three immunomodulatory molecules are OX40 Ligand (OX40L), CD27 Ligand (CD70) and CD28 Ligand (CD28L) comprising CD80, CD86, or both, for induction of one or more subpopulations of PBMCs to proliferate in response to the expressed immunomodulatory molecules and to then enter an effector phase for killing of tumor cells;   wherein the subpopulations of PBMC cells comprise one or more of T-lymphocytes, natural killer (NK) cells, dendritic cells (DCs) or B lymphocytes; and   (2) a pharmaceutically acceptable carrier.   
     
     
         2 . The allogeneic tumor cell vaccine of  claim 1 , wherein
 (a) the population of live, proliferation-incompetent genetically engineered tumor cells expressing one or more tumor specific antigens further comprises one or more additional stably expressed immunomodulatory molecules selected from R 1 -R 44 ; or   (b) the tumor cells are rendered proliferation-incompetent by irradiation; or   (c) the tumor cell is derived from a cancer selected from the group consisting of:   melanoma, colorectal carcinoma, leukemia, chronic myeloid leukemia, prostate cancer, head and neck cancer, Squamous Cell Carcinoma, tongue cancer, larynx cancer, tonsil cancer, hypopharynx cancer, nasalpharynx cancer, breast cancer, colon cancer, lung cancer, pancreatic cancer, glioblastoma and brain cancer; or   (d) the population of live, proliferation-resistant tumor cells is derived from a biological sample derived from a subject; or   (e) the population of live proliferation resistant tumor cells is derived from a tumor cell line; or   (f) the immunostimulatory molecules are presented at the exterior surface of the genetically engineered tumor cells; or   (g) induction of the T-lymphocytes comprises activation of the subpopulation of T lymphocytes, expansion of the T lymphocytes, or both; or   (h) induction of the NK cells comprises activation of the subpopulation of NK cells, expansion of the subpopulation of NK cells or both; or   (i) induction of the subpopulation of DCs comprises activation of the subpopulation of DCs, expansion of the subpopulation of DCs or both; or   (j) induction of the subpopulation of B lymphocytes comprises activation of the subpopulation of B lymphocytes, expansion of the subpopulation of B lymphocytes or both.   
     
     
         3 . The allogeneic tumor cell vaccine of  claim 1 , wherein
 (a) the subpopulation of T lymphocytes comprises a subpopulation of CD8+ cytotoxic T-lymphocytes (CTL); or   (b) the subpopulation of T lymphocytes comprises a subpopulation of memory T cells; or   (c) the subpopulation of T lymphocytes comprises a subpopulation of regulatory T cells; or   (d) the subpopulation of T lymphocytes comprises a subpopulation of helper T cells; or   (e) the subpopulation of B lymphocytes comprises a subpopulation of memory B cells.   
     
     
         4 . The allogeneic tumor cell vaccine of  claim 1 , wherein:
 (1) the vaccine enhances immune activation of cells effective to recognize and act against those tumor cells that comprise the target tumor antigen in vivo without systemic inflammation; or   (2) the vaccine reduces immunosuppression in a tumor microenvironment for tumor cells comprising the target tumor antigen; or   (3) the vaccine increases cell death of tumor cells expressing the target tumor antigen; or   (4) the population of live, proliferation-resistant tumor cells elicits immune activation without systemic inflammation; or   (5) the vaccine elicits an immune response that improves progression free survival, overall survival, or both relative to placebo controls.   
     
     
         5 . The allogeneic tumor cell vaccine of  claim 2 , wherein the melanoma tumor cell is characterized by expression of one or more of gp100, tyrosinase, Melan-A, tyrosinase-related protein (TRP-2-INT2), melanoma antigen-1 (MAGE-A1), NY-ESO-1, preferentially expressed antigen of melanoma (PRAME) CDK4 and multiple myeloma oncogene 1 (MUM-1). 
     
     
         6 . The allogeneic tumor cell vaccine of  claim 2 , wherein the colorectal cancer tumor cell is characterized by expression of one or more of carcinoembryonic antigen (CEA), MAGE, HPV, human telomerase reverse transcriptase (hTERT), EPCAM, PD-1, PD-L1, p53, and cell surface-associated mucin 1 (MUC1). 
     
     
         7 . The allogeneic tumor cell vaccine of  claim 2 , wherein the one or more additional stably expressed immunomodulatory molecules selected from R 1 -R 44  is a cytokine, a TNF-family member, a secreted receptor, a chaperone, an IgG superfamily member and/or a chemokine receptor. 
     
     
         8 . The allogeneic tumor cell vaccine of  claim 1 , produced by a process comprising:
 providing an allogeneic parental tumor cell line comprising a population of live tumor cells;   introducing into the population of live tumor cells an exogenous nucleic acid encoding a stably expressed immunomodulatory molecule, wherein the immunomodulatory molecule is OX40 Ligand (OX40L);   introducing into the population of live tumor cells an exogenous nucleic acid encoding a stably expressed immunomodulatory molecule, wherein the immunomodulatory molecule is CD27 Ligand (CD70);   introducing into the population of live tumor cells an exogenous nucleic acid encoding a stably expressed immunomodulatory molecule, wherein the immunomodulatory molecule is CD28 Ligand (CD28L) comprising CD80, CD86, or both; wherein stable expression of OX40 Ligand (OX40L), CD27 Ligand (CD70) and CD28 Ligand (CD28L) comprising CD80, CD86, or both induces one or more subpopulations of PBMCs to proliferate in response to the expressed immunomodulatory molecules and to then enter an effector phase for killing of tumor cells;   generating tumor cell line variants by selecting for tumor cell clones that stably express an immunogenic amount of the exogenous subset of the immunomodulatory molecules; and   selecting in a mixed lymphocyte tumor cell reaction clonally derived cell line variants by one or more of the following parameters selected from: cellular proliferation, cellular subset differentiation, cytokine release profile, and tumor cell lysis; wherein the selected clonally derived cell line variant is effective to stimulate activation of one or more of T-lymphocytes, natural killer (NK) cells, dendritic cells (DCs) or B lymphocytes.   
     
     
         9 . The allogeneic tumor cell vaccine produced by the process of  claim 8 , further comprising introducing into the population of live tumor cells an exogenous nucleic acid encoding one or more stably expressed immunomodulatory molecules selected from R 1 -R 44 . 
     
     
         10 . The allogeneic tumor cell vaccine produced by the process of  claim 8 , wherein
 (a) the tumor cells are rendered proliferation incompetent by irradiation; or   (b) the parental tumor cell line is from a melanoma or from a colorectal carcinoma; or   (c) the exogenous nucleic acid comprises DNA or RNA; or   (d) the introducing step comprises viral transduction; or   (e) the introducing step comprises electroporation; or   (f) the introducing step comprises utilizing one or more of: liposome mediated transfer, adenovirus, adeno-associated virus, herpes virus, a retroviral based vector, lipofection, and a lentiviral vector; or   (g) the introducing step comprises introducing the exogenous nucleic acid by transfection of a lentiviral vector.   
     
     
         11 . A method of inducing an immune response to a cancer in a subject comprising:
 (a) administering to the subject parenterally or locally into a tumor an allogeneic tumor cell vaccine comprising (1) a population of live, proliferation-incompetent genetically engineered tumor cells expressing one or more tumor specific antigens, the population comprising at least three stably expressed immunomodulatory molecules, wherein the at least three immunomodulatory molecules are OX40 Ligand (OX40L), CD27 Ligand (CD70) and CD28 Ligand (CD28L) comprising, wherein CD28 Ligand comprises CD80, CD86, or both, and (2) a pharmaceutically acceptable carrier,   (b) inducing one or more subpopulations of peripheral blood mononuclear cells (PBMCs) to proliferate in response to the expressed immunomodulatory molecules and to then enter an effector phase for killing of tumor cells;
 wherein the subpopulations of PBMC cells comprise one or more of T-lymphocytes, natural killer (NK) cells, dendritic cells (DCs) or B lymphocytes; and 
 wherein the allogeneic tumor cell vaccine is type-matched to the subject's cancer. 
   
     
     
         12 . The method of  claim 11 , wherein the cancer is selected from melanoma or colorectal cancer. 
     
     
         13 . The method of  claim 11 , wherein the subject has an infectious viral disease with progression to a cancer. 
     
     
         14 . The method of  claim 11 , further comprising administering a checkpoint inhibitor to the subject. 
     
     
         15 . The method of  claim 11 , wherein the population of live, proliferation-incompetent genetically engineered tumor cells expressing one or more tumor specific antigens further comprises one or more additional stably expressed immunomodulatory molecules selected from R 1 -R 44 . 
     
     
         16 . The method according to  claim 11 , wherein
 (a) the population of live, proliferation-resistant tumor cells is derived from a biological sample derived from a subject; or   (b) the population of live proliferation resistant tumor cells is derived from a tumor cell line.   
     
     
         17 - 22 . (canceled)