Enhanced chimeric antigen receptors and use thereof

Abstract

Provided herein are chimeric antigen receptors (CARs) comprising a truncated EGFRvIII (Ev3) in the hinge region of the CAR and/or a humanized scFv. Further provided herein are immune cells expressing the CARs as well as methods of their use in the treatment of immune disorders.

What is claimed is: 
     
         1 . An isolated antigen-specific humanized single chain variable fragment (scFv) comprising a light chain variable region (V L ) and a heavy chain variable region (V H ), wherein the scFv is CD5-specific and wherein the V L  comprises an amino acid sequence of SEQ ID NO:31 and the V H  comprises an amino acid sequence of SEQ ID NO:32, or sequences with 90% sequence identity to framework regions of SEQ ID NO:31 and SEQ ID NO:32, respectively. 
     
     
         2 . The humanized scFv of  claim 1 , wherein the CD5-specific scFv comprises an amino acid sequence of SEQ ID NO:30. 
     
     
         3 . The humanized scFv of  claim 1 , wherein the scFv comprises an amino acid sequence with at least 95% sequence identity to framework regions in an amino acid sequence of SEQ ID NO: 30. 
     
     
         4 . The humanized scFv of  claim 1 , wherein the scFv is comprised in a chimeric antigen receptor (CAR). 
     
     
         5 . The humanized scFv of  claim 4 , wherein the CAR comprises at least one signaling domain selected from the group consisting of CD3, CD28, OX40/CD134, 4-1BB/CD137, FcεRIγ, ICOS/CD278, ILRB/CD122, IL-2RG/CD132, DAP10, DAP12, CD70, CD40, and a combination thereof. 
     
     
         6 . The humanized scFv of  claim 5 , wherein the at least one signaling domain comprises DAP10 or DAP12. 
     
     
         7 . The humanized scFv of  claim 4 , wherein the CAR is co-expressed with IL-15. 
     
     
         8 . An isolated polynucleotide encoding the isolated humanized scFv of  claim 1 . 
     
     
         9 . The polynucleotide of  claim 8 , wherein the polynucleotide comprises nucleotide sequence encoding a CAR that comprises the scFv. 
     
     
         10 . An expression vector comprising the isolated polynucleotide of  claim 9 . 
     
     
         11 . The vector of  claim 10 , wherein the vector expresses the CAR and expresses IL-15. 
     
     
         12 . The vector of  claim 10 , wherein the vector is further defined as a viral vector. 
     
     
         13 . A host cell engineered to express the polynucleotide of  claim 9 . 
     
     
         14 . The cell of  claim 13 , wherein the host cell is further defined as an immune cell. 
     
     
         15 . The cell of  claim 14 , wherein the immune cell is a T cell, peripheral blood lymphocyte, NK cell, invariant NK cell, NKT cell, or stem cell. 
     
     
         16 . The cell of  claim 14 , wherein the immune cell is isolated from cord blood. 
     
     
         17 . A pharmaceutical composition comprising a population of cells according to  claim 13 . 
     
     
         18 . A method of treating an immune-related disorder in a subject comprising administering an effective amount of cells of  claim 13  to the subject. 
     
     
         19 . The method of  claim 18 , wherein the immune-related disorder is a cancer, autoimmune disorder, graft versus host disease, allograft rejection, or inflammatory condition.