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 method of treating cancer, comprising administering an immune cell to a human subject having cancer, said immune cell 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 associated with the cancer, a transmembrane domain, and an intracellular signal transduction domain. 
     
     
         151 . The method of  claim 150 , wherein the target antigen is a cancer antigen whose expression is increased in or on the surface of a cancer cell, a cancer tissue, and/or a tumor microenvironment. 
     
     
         152 . The method of  claim 150 , wherein the target antigen is CD19 or CD22. 
     
     
         153 . The method of  claim 150 , wherein the immune cell is a human-derived T cell, a dendritic cell or natural killer (NK) cell. 
     
     
         154 . The method of  claim 150 , wherein the immune cell is a human-derived T cell. 
     
     
         155 . The method 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 (4-1BB), CD27, and CD28. 
     
     
         156 . The method of  claim 150 , wherein the first nucleotide sequence is present on a vector. 
     
     
         157 . The method of  claim 150 , wherein the first nucleotide sequence and the second nucleotide sequence are present on a single vector. 
     
     
         158 . A vector comprising (i) a first nucleotide sequence encoding a first 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 CAR. 
     
     
         159 . The vector of  claim 158 , wherein the expression of the first shRNA is regulated by a first promoter and the second shRNA is regulated by a second promoter. 
     
     
         160 . The vector of  claim 159 , wherein the first and the second promoters are RNA polymerase III promoters. 
     
     
         161 . The vector of  claim 160 , wherein the RNA polymerase III promoters are U6 promoters. 
     
     
         162 . The vector of  claim 159 , wherein the two promoters are oriented in different directions from each other on the vector. 
     
     
         163 . The vector of  claim 162 , wherein the two promoters are oriented in head to head orientation. 
     
     
         164 . The vector of  claim 162 , wherein the two promoters are oriented in tail to tail orientation. 
     
     
         165 . An immune cell comprising the vector of  claim 158 . 
     
     
         166 . A pharmaceutical composition comprising the immune cell of  claim 165  and a pharmaceutically acceptable carrier.