Intracellular genomic transplant and methods of therapy

Abstract

Genetically modified compositions, such as non-viral vectors and T cells, for treating cancer are disclosed. Also disclosed are the methods of making and using the genetically modified compositions in treating cancer.

1 .- 150 . (canceled) 
     
     
         151 . A pharmaceutical composition comprising:
 (a) population of genetically modified immune cells comprising:
 (i) a genomic disruption in a cytokine inducible SH2-containing protein (CISH) gene target sequence; and 
 (ii) an exogenous chimeric antigen receptor (CAR) or an exogenous T-cell receptor (TCR), wherein the exogenous CAR or the exogenous TCR target a tumor cell antigen, wherein the tumor cell antigen is a MUC-1 antigen or a CD70 antigen, 
   wherein the population of genetically modified immune cells comprise a population of T cells, a population of NK cells, or a combination thereof, and   (b) a pharmaceutically acceptable carrier or excipient.   
     
     
         152 . The pharmaceutical composition of  claim 151 , wherein the exogenous CAR or the exogenous TCR are encoded by an exogenous nucleic acid, wherein the exogenous nucleic acid is inserted into an endogenous TCR gene. 
     
     
         153 . The pharmaceutical composition of  claim 151 , wherein the exogenous CAR or the exogenous TCR are encoded by an exogenous nucleic acid, wherein the exogenous nucleic acid is inserted adjacent to, near, or within the CISH target sequence. 
     
     
         154 . The pharmaceutical composition of  claim 151 , wherein the genomic disruption is an endonuclease-mediated indel within 5 base pairs of a protospacer adjacent motif (PAM) sequence and a disruption efficiency of at least 30% 
     
     
         155 . The pharmaceutical composition of  claim 151 , wherein the genetically engineered population of immune cells are cluster of differentiation 56 positive (CD56*) cells. 
     
     
         156 . The pharmaceutical composition of  claim 151 , wherein the population of genetically modified immune cells comprise a population of autologous cells derived from a human subject in need of treatment with the pharmaceutical composition. 
     
     
         157 . The pharmaceutical composition of  claim 151 , wherein the population of genetically modified immune cells comprise a population of immune cells derived from pluripotent stem cells. 
     
     
         158 . The pharmaceutical composition of  claim 154 , wherein the disruption efficiency of the CISH gene target sequence is determined by tracking of indels by decomposition (TIDE) analysis. 
     
     
         159 . A pharmaceutical compositions comprising:
 (a) population of genetically modified natural killer (NK) cells comprising:   a genomic disruption in a cytokine inducible SH2-containing protein (CISH) gene target sequence and at least one additional genomic disruption in a transforming growth factor beta receptor II (TGFBRII) gene target sequence; and   (b) a pharmaceutically acceptable carrier or excipient.   
     
     
         160 . The pharmaceutical composition of  claim 159 , wherein the genetically engineered population of NK cells are CD56+ cells. 
     
     
         161 . The pharmaceutical composition of  claim 159 , wherein the population of genetically modified NK cells further comprise an exogenous CAR or an exogenous TCR, wherein the exogenous CAR or the exogenous TCR target a tumor antigen. 
     
     
         162 . The pharmaceutical composition of  claim 161 , wherein the exogenous CAR or the exogenous TCR are encoded by an exogenous nucleic acid. 
     
     
         163 . The pharmaceutical composition of  claim 161 , wherein the exogenous CAR or the exogenous TCR are encoded by an exogenous nucleic acid, wherein the exogenous nucleic acid is inserted adjacent to, near, or within the CISH target sequence or the TGFBRII target sequence. 
     
     
         164 . The pharmaceutical composition of  claim 159 , wherein the population of genetically modified NK cells comprise a population of autologous cells derived from a human subject in need of treatment with the pharmaceutical composition. 
     
     
         165 . The pharmaceutical composition of  claim 159 , wherein the population of genetically modified NK cells comprise a population of NK cells derived from pluripotent stem cells. 
     
     
         166 . The pharmaceutical composition of  claim 159 , wherein at least one genomic disruption is an endonuclease-mediated indel within 5 base pairs of a protospacer adjacent motif (PAM) sequence and a disruption efficiency of at least 30%. 
     
     
         167 . The pharmaceutical composition of  claim 159 , wherein the disruption efficiency of at least one gene target sequence is determined by tracking of indels by decomposition (TIDE) analysis.