Combinatorial cancer immunotherapy

Abstract

Provided herein are methods and compositions for dynamically controlling and targeting multiple immunosuppressive mechanisms in cancer. Some aspects provide cells engineered to produce multiple effector molecules, each of which modulates a different immunosuppressive mechanisms of a tumor, as well as methods of using the cells to treat cancer, such as ovarian, breast, or colon cancer.

1 . A method of reducing tumor volume in a subject in need thereof, the method comprising delivering to the subject having a tumor a composition comprising natural killer (NK) cells engineered to produce multiple effector molecules comprising at least a first effector molecule and a second effector molecule that each modulate tumor-mediated immunosuppressive mechanisms in an effective amount to reduce the volume of the tumor, wherein the first effector molecule comprises interleukin 12 (IL-12),
 wherein the multiple effector molecules except the first effector molecule are each independently selected from the group consisting of: cytokines, receptors/ligands, antibodies, nucleotides, peptides, and enzymes,   wherein the composition comprises (a) a first NK cell engineered to produce the first effector molecule and (b) a second NK cell engineered to produce the second effector molecule, and   wherein tumor reduction is greater than tumor reduction following administration of either the first NK cell alone or the second NK cell alone.   
     
     
         2 . The method of  claim 1 , wherein the tumor is selected from the group consisting of: bladder tumors, brain tumors, breast tumors, cervical tumors, colorectal tumors, esophageal tumors, gliomas, kidney tumors, liver tumors, lung tumors, melanomas, ovarian tumors, pancreatic tumors, prostate tumors, skin tumors, thyroid tumors, and uterine tumors. 
     
     
         3 . The method of  claim 1 , wherein the second effector molecule is selected from the group consisting of: MIP1a (CCL3), MIP1b (CCL5), CCL21, CXCL9, CXCL10 CXCL11, IFN-γ, IL-2, IL-4, IL-15, IL-16, IL-7, IL-9, IL-36γ, IL-18, IL-21, IL-10, OX40-ligand, CD40L, an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody, CCR9, CXCR3, CXCR4, CCR2, CCR4, FPR2, VEGFR, IL6R, CXCR1, CSCR7, and PDGFR. 
     
     
         4 . The method of  claim 1 , wherein the method further comprises delivering to the subject a checkpoint inhibitor. 
     
     
         5 . The method of  claim 5 , wherein the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-1L antibody, or an anti-CTLA-4 antibody. 
     
     
         6 . The method of  claim 5 , wherein the checkpoint inhibitor is an anti-CTLA-4 antibody. 
     
     
         7 . The method of  claim 1 , wherein the method further comprises delivering to the subject an anti-CD40 antibody. 
     
     
         8 . The method of  claim 1 , wherein the volume of the tumor is reduced by at least 25% relative to a control. 
     
     
         9 . The method of  claim 1 , wherein the second effector molecule comprises IL-21. 
     
     
         10 . The method of  claim 1 , wherein the second effector molecule comprises IL-15. 
     
     
         11 . The method of  claim 1 , wherein the composition is administered intravenously. 
     
     
         12 . The method of  claim 1 , wherein the tumor is a colorectal tumor or a liver tumor. 
     
     
         13 . The method of  claim 1 , wherein the tumor reduction is equal to or greater than the sum of the tumor reduction following separate administration of the first NK cell and the second NK cell. 
     
     
         14 . A method of stimulating an immune response in a subject having a tumor, the method comprising delivering to the subject a composition comprising NK cells engineered to produce multiple effector molecules comprising at least a first effector molecule and a second effector molecule that each stimulate the immune response in an effective amount to stimulate the immune response, wherein the first effector molecule comprises IL-12,
 wherein stimulating the immune response comprises increasing an immune response in the subject, thereby reducing tumor volume,   wherein the multiple effector molecules except the first effector molecule are each independently selected from the group consisting of: cytokines, receptors/ligands, antibodies, nucleotides, peptides, and enzymes,   wherein the composition comprises (a) a first NK cell engineered to produce the first effector molecule and (b) a second NK cell engineered to produce the second effector molecule, and   wherein the increased immune response is greater than the immune response following administration of either the first NK cell alone or the second NK cell alone.   
     
     
         15 . The method of  claim 14 , wherein the subject has a tumor selected from the group consisting of: bladder tumors, brain tumors, breast tumors, cervical tumors, colorectal tumors, esophageal tumors, gliomas, kidney tumors, liver tumors, lung tumors, melanomas, ovarian tumors, pancreatic tumors, prostate tumors, skin tumors, thyroid tumors, and uterine tumors. 
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 14 , wherein the second effector molecule comprises IL-21. 
     
     
         18 . The method of  claim 14 , wherein the second effector molecule comprises IL-15. 
     
     
         19 . The method of  claim 14 , wherein the composition is administered intravenously. 
     
     
         20 . The method of  claim 14 , wherein the tumor volume reduction is equal to or greater than the sum of the tumor volume reduction following separate administration of the first NK cell and the second NK cell. 
     
     
         21 . A composition comprising natural killer (NK) cells or MSCs engineered to produce multiple effector molecules comprising at least a first effector molecule and a second effector molecule that each modulate tumor-mediated immunosuppressive mechanisms, wherein the first effector molecule comprises interleukin 12 (IL-12),
 wherein the multiple effector molecules except the first effector molecule are each independently selected from the group consisting of: cytokines, receptors/ligands, antibodies, nucleotides, peptides, and enzymes, and   wherein the composition comprises (a) a first NK or MSC cell engineered to produce the first effector molecule and (b) a second NK or MSC cell engineered to produce the second effector molecule.