Enhanced immune cells using dual shrna and composition including the same

Abstract

The present disclosure is broadly concerned with the field of cancer immunotherapy. For example, the present invention generally relates to an immune cell comprising a genetically engineered antigen receptor that specifically binds to a target antigen and a genetic disruption agent that reduces or is capable of reducing the expression in the immune cell of a gene that weakens the function of the immune cell.

1 - 149 . (canceled) 
     
     
         150 . A vector comprising:
 (i) a first nucleotide sequence encoding a first short hairpin RNA (shRNA) that inhibits expression of programmed cell death 1 (PD-1) and a second shRNA that inhibits expression of T cell immunoreceptor with Ig and ITIM domains (TIGIT); and   (ii) a second nucleotide sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an extracellular antigen recognition domain that binds a target antigen, a transmembrane domain, and an intracellular signal transduction domain.   
     
     
         151 . The vector of  claim 150 , wherein the target antigen is a cancer antigen whose expression is increased on the surface of a cancer cell, in a cancer tissue, and/or in a tumor microenvironment. 
     
     
         152 . The vector of  claim 151 , wherein the cancer antigen is CD19 or CD22. 
     
     
         153 . The vector of  claim 151 , wherein the cancer antigen is associated with a B-cell blood cancer. 
     
     
         154 . The vector of  claim 153 , wherein the B cell blood cancer is B-cell lymphoma. 
     
     
         155 . The vector of  claim 150 , wherein the intracellular signal transduction domain of the CAR further comprises a costimulatory molecule selected from the group consisting of ICOS, OX40, CD137 (4-IBB), CD27, and CD28. 
     
     
         156 . The vector of  claim 150 , wherein the first nucleotide sequence and the second nucleotide sequence are present on a single vector. 
     
     
         157 . The vector of  claim 158 , wherein expression of the first shRNA is regulated by a first promoter and expression of the second shRNA is regulated by a second promoter. 
     
     
         158 . The vector of  claim 159 , wherein the first promoter and the second promoter are oriented in different directions from each other on the single vector. 
     
     
         159 . The vector of  claim 160 , wherein the first promoter and the second promoter are in a head-to-head orientation. 
     
     
         160 . The vector of  claim 160 , wherein the first promoter and the second promoter are in a tail-to-tail orientation. 
     
     
         161 . The vector of  claim 159 , wherein the first promoter and the second promoter are RNA polymerase III promoters. 
     
     
         162 . An immune cell comprising a vector comprising:
 (i) a first nucleotide sequence encoding a first short hairpin RNA (shRNA) that inhibits expression of programmed cell death 1 (PD-1) and a second shRNA that inhibits expression of T cell immunoreceptor with Ig and ITIM domains (TIGIT); and   (ii) a second nucleotide sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an extracellular antigen recognition domain that binds a target antigen, a transmembrane domain, and an intracellular signal transduction domain.   
     
     
         163 . The immune cell of  claim 162 , wherein the target antigen is a cancer antigen whose expression is increased on the surface of a cancer cell, in a cancer tissue, and/or in a tumor microenvironment. 
     
     
         164 . The immune cell of  claim 163 , wherein the cancer antigen is CD19 or CD22. 
     
     
         165 . The immune cell of  claim 163 , wherein the cancer antigen is associated with a B-cell blood cancer. 
     
     
         166 . The immune cell of  claim 165 , wherein the B cell blood cancer is B-cell lymphoma. 
     
     
         167 . The immune cell of  claim 162 , wherein the immune cell is a human derived T cell, dendritic cell, or natural killer (NK) cell. 
     
     
         168 . The immune cell of  claim 167 , wherein the immune cell is a human derived T cell. 
     
     
         169 . The immune cell of  claim 162 , wherein the intracellular signal transduction domain of the CAR further comprises a costimulatory molecule selected from the group consisting of ICOS, OX40, CD137 (4-IBB), CD27, and CD28. 
     
     
         170 . The immune cell of  claim 162 , wherein the first nucleotide sequence and the second nucleotide sequence are present on a single vector. 
     
     
         171 . The immune cell of  claim 170 , wherein expression of the first shRNA is regulated by a first promoter and expression of the second shRNA is regulated by a second promoter. 
     
     
         172 . The immune cell of  claim 171 , wherein the first promoter and the second promoter are oriented in different directions from each other on the single vector. 
     
     
         173 . The immune cell of  claim 172 , wherein the first promoter and the second promoter are in a head-to-head orientation. 
     
     
         174 . The immune cell of  claim 172 , wherein the first promoter and the second promoter are in a tail-to-tail orientation. 
     
     
         175 . The immune cell of  claim 171 , wherein the first promoter and the second promoter are RNA polymerase III promoters. 
     
     
         176 . A pharmaceutical composition comprising an immune cell and a pharmaceutically acceptable carrier, wherein the immune cell comprises:
 (i) a first nucleotide sequence encoding a first short hairpin RNA (shRNA) that inhibits expression of programmed cell death 1 (PD-1) and a second shRNA that inhibits expression of T cell immunoreceptor with Ig and ITIM domains (TIGIT); and   (ii) a second nucleotide sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an extracellular antigen recognition domain that binds a target antigen, a transmembrane domain, and an intracellular signal transduction domain.   
     
     
         177 . A method of treating cancer, comprising administering an immune cell to a human subject having cancer, wherein the immune cell comprises:
 (i) a first nucleotide sequence encoding a first short hairpin RNA (shRNA) that inhibits expression of programmed cell death 1 (PD-1) and a second shRNA that inhibits expression of T cell immunoreceptor with Ig and ITIM domains (TIGIT); and   (ii) a second nucleotide sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an extracellular antigen recognition domain that binds a target antigen, a transmembrane domain, and an intracellular signal transduction domain.