Affinity-enhanced monmeric streptavidin chimeric antigen receptor (car)

Abstract

A chimeric antigen receptor is disclosed that includes: (a) an extracellular high affinity streptavidin; (b) a hinge domain from CD8; (c) a CD28 transmembrane domain; (d) an intracellular 4-1BB and/or CD28 signaling domain; and (e) an intracellular CD3 zeta signaling domain, wherein (a)-(e) are in N-terminal to C-terminal order. Nucleic acids encoding this chimeric antigen receptor, and T and natural killer (NK) cells transformed with this chimeric antigen receptor are also disclosed. The use of this chimeric antigen receptor for the treatment of tumors is also disclosed.

We claim: 
     
         1 . An isolated nucleic acid molecule encoding a chimeric antigen receptor, wherein the chimeric antigen receptor comprises amino acids 1-369 of SEQ ID NO: 11, or amino acids 1-371 of SEQ ID NO: 12. 
     
     
         2 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleic acid sequence of one or more of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19, and a degenerate variant thereof. 
     
     
         3 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleic acid sequence of one or more of SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19. 
     
     
         4 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, or SEQ ID NO: 23, or a degenerate variant thereof. 
     
     
         5 . The isolated nucleic acid molecule of  claim 1 , comprising the nucleic acid of SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, or SEQ ID NO: 23. 
     
     
         6 . The isolated nucleic acid molecule of  claim 1 , wherein the nucleic acid molecule is codon-optimized for expression in human cells. 
     
     
         7 . The isolated nucleic acid molecule of  claim 1 , operably linked to a promoter. 
     
     
         8 . An expression vector comprising the nucleic acid molecule of  claim 1 . 
     
     
         9 . The expression vector of  claim 8 , wherein the expression vector is a viral vector. 
     
     
         10 . The expression vector of  claim 9 , wherein the viral vector is a lentiviral vector or a gamma retroviral vector. 
     
     
         11 . The expression vector of  claim 10 , wherein the viral vector is the gamma retroviral vector. 
     
     
         12 . An isolated host cell comprising the vector of  claim 8 . 
     
     
         13 . The isolated host cell of  claim 12 , wherein the host cell is a CD3+ T cell or a natural killer cell. 
     
     
         14 . The isolated host cell of  claim 13 , wherein the host cell is a human cell. 
     
     
         15 . The isolated host cell of  claim 13 , wherein the CD3+ T cell is a CD3 + CD4 +  T cell or CD3 + CD8 +  T cell. 
     
     
         16 . A pharmaceutical composition comprising the nucleic acid molecule of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         17 . A pharmaceutical composition comprising the expression vector of  claim 8  and a pharmaceutically acceptable carrier.