Chimeric antigen receptor and natural killer cells expressing same

Abstract

The present invention provides a chimeric antigen receptor and natural killer cells expressing the same, and particularly, a chimeric antigen receptor (CAR) which includes an intracellular signaling domain including the whole or a portion of an OX40 ligand (CD252), thereby having excellent effects of increasing anticancer activity of immune cells, and immune cells expressing the same.

1 - 30 . (canceled) 
     
     
         31 . A chimeric antigen receptor, comprising:
 an intracellular signaling domain which includes the whole or a portion of OX40 ligand (CD252).   
     
     
         32 . The chimeric antigen receptor according to  claim 31 , further comprising:
 a transmembrane domain linked to the intracellular signaling domain;   a spacer domain linked to the transmembrane domain; and   an extracellular domain linked to the spacer domain.   
     
     
         33 . The chimeric antigen receptor according to  claim 32 , further comprising a signal sequence linked to the extracellular domain. 
     
     
         34 . The chimeric antigen receptor according to  claim 33 , wherein the signal sequence includes the whole or a portion of CD16 and the signal sequence includes the whole or a portion of CD8α, respectively. 
     
     
         35 . The chimeric antigen receptor according to  claim 32 , wherein the extracellular domain includes the whole or a portion of any one selected from the group consisting of an antigen-binding fragment of an antibody, a Fc receptor, a natural cytotoxicity receptor, NKG2D, 2B4 and DNAM-1. 
     
     
         36 . The chimeric antigen receptor according to  claim 35 , wherein the antigen-binding fragment is Fab fragment, F(ab′) fragment, F(ab′)2 fragment or Fv fragment. 
     
     
         37 . The chimeric antigen receptor according to  claim 36 , wherein the Fv fragment is a single-chain variable fragment (ScFv). 
     
     
         38 . The chimeric antigen receptor according to  claim 35 , wherein the Fc receptor is selected from the group consisting of CD16, CD32, CD64, CD23, CD89 and variants thereof. 
     
     
         39 . The chimeric antigen receptor according to  claim 38 , wherein the Fc receptor is CD16 or variants thereof. 
     
     
         40 . The chimeric antigen receptor according to  claim 35 , wherein the natural cytotoxicity receptor is selected from the group consisting of NKp46, NKp30, NKp44, NKp80 and NKp65 receptors. 
     
     
         41 . The chimeric antigen receptor according to  claim 32 , wherein the spacer domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28. 
     
     
         42 . The chimeric antigen receptor according to  claim 32 , wherein the transmembrane domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28. 
     
     
         43 . The chimeric antigen receptor according to  claim 31 , wherein the intracellular signaling domain further includes the whole or a portion of CD3-zeta. 
     
     
         44 . The chimeric antigen receptor according to  claim 43 , wherein the whole or a portion of CD3-zeta and the whole or a portion of OX40 ligand are arranged in order from a cell membrane toward an inside of the cell. 
     
     
         45 . The chimeric antigen receptor according to  claim 32 , further comprising an AAA linker between the extracellular domain and the spacer domain. 
     
     
         46 . A chimeric antigen receptor, comprising intracellular signaling domains which comprise:
 a first intracellular signaling domain including the whole or a portion of any one selected from the group consisting of CD28 and 4-1BB;   a second intracellular signaling domain including the whole or a portion of any one selected from the group consisting of OX40 ligand, OX40 and 4-1BB; and   a third intracellular signaling domain including the whole or a portion of CD3-zeta,   wherein the first, second and third intracellular signaling domains are arranged in order from a cell membrane toward an inside of the cell.   
     
     
         47 . The chimeric antigen receptor according to  claim 46 , further comprising:
 a transmembrane domain linked to the intracellular signaling domain;   a spacer domain linked to the transmembrane domain; and   an extracellular domain linked to the spacer domain.   
     
     
         48 . The chimeric antigen receptor according to  claim 47 , further comprising a signal sequence linked to the extracellular domain. 
     
     
         49 . The chimeric antigen receptor according to  claim 48 , wherein the signal sequence includes the whole or a portion of CD16 and the signal sequence includes the whole or a portion of CD8α, respectively. 
     
     
         50 . The chimeric antigen receptor according to  claim 47 , wherein the extracellular domain includes the whole or a portion of any one selected from the group consisting of an antigen-binding fragment of an antibody, a Fc receptor, a natural cytotoxicity receptor, NKG2D, 2B4 and DNAM-1. 
     
     
         51 . The chimeric antigen receptor according to  claim 47 , wherein the spacer domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28. 
     
     
         52 . The chimeric antigen receptor according to  claim 47 , wherein the transmembrane domain includes the whole or a portion of any one selected from the group consisting of CD8α and CD28. 
     
     
         53 . The chimeric antigen receptor according to  claim 46 , wherein the first intracellular signaling domain includes the whole or a portion of CD28;
 the second intracellular signaling domain includes the whole or a portion of the OX40 ligand; and   the third intracellular signaling domain includes the whole or a portion of CD3-zeta.   
     
     
         54 . The chimeric antigen receptor according to  claim 47 , further comprising an AAA linker between the extracellular domain and the spacer domain. 
     
     
         55 . An immune cell expressing the chimeric antigen receptor according to  claim 31 . 
     
     
         56 . The immune cell according to  claim 55 , wherein the immune cell is a natural killer cell (NK cell). 
     
     
         57 . A pharmaceutical composition for treatment of tumor, comprising the immune cell according to  claim 55  as an active ingredient. 
     
     
         58 . The pharmaceutical composition according to  claim 57 , further comprising an antibody as an active ingredient when the extracellular domain is a Fc receptor. 
     
     
         59 . A nucleic acid sequence encoding the chimeric antigen receptor according to  claim 31 . 
     
     
         60 . The nucleic acid sequence according to  claim 59 , wherein the nucleic acid sequence encodes one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 33, 41, 43, 45, 47, 49, 51, 53, 55, 69, 71, 77, 81, 83, 85, 87, 89, 91 and 93, or variants thereof having a sequence identity of 80% or more. 
     
     
         61 . The nucleic acid sequence according to  claim 60 , wherein the nucleic acid sequence includes one or more nucleic acid sequence selected from the group consisting of SEQ ID NOs: 32, 40, 42, 44, 46, 48, 50, 52, 54, 68, 70, 76, 80, 82, 84, 86, 88, 90 and 92, or variants thereof having a sequence identity of 80% or more. 
     
     
         62 . A method for treatment of tumor, comprising administering the immune cell according to  claim 55  to a subject.