Inhibitory chimeric antigen receptors

Abstract

The invention relates to an inhibitory chimeric antigen receptor (N-CAR) comprisingan extracellular domain comprising an antigen binding domain,a transmembrane domain, and,an intracellular domainwherein the intracellular domain comprises an Immunoreceptor Tyrosine-based Switch Motif ITSM, wherein said ITSM is a sequence of amino acid TX1YX2X3X4, whereinX1 is an amino acidX2 is an amino acidX3 is an amino acid andX4 is V or I.

1 . A method for treating a patient in need thereof comprising: (a) providing an immune cell; and (b) administering said immune cell to said patient, wherein the immune cell comprises:
 a first CAR comprising an extracellular domain comprising an antigen binding domain, a transmembrane domain, and an intracellular domain; and   a second CAR comprising an extracellular domain comprising an antigen binding domain, a transmembrane domain, and an intracellular domain, wherein:   the intracellular domain of the second CAR comprises an Immunoreceptor Tyrosine-based Switch Motif (ITSM), wherein said ITSM is a sequence of amino acids TX 1 YX 2 X 3 X 4  (SEQ ID NO: 2049), wherein
 X 1  is E; 
 X 2  is A or S; 
 X 3  is S or E; and 
 X 4  is V or I; and 
    the intracellular domain of the second CAR has at least 95% amino acid sequence identity with SEQ ID NO: 2016.   
     
     
         2 . The method of  claim 1 , wherein the ITSM is selected from the group consisting of TEYASI (SEQ ID NO: 936), TEYSEI (SEQ ID NO: 940), and TEYSEV (SEQ ID NO: 941). 
     
     
         3 . The method of  claim 1 , wherein the antigen binding domain of the second CAR is a single chain variable fragment (scFv). 
     
     
         4 . The method of  claim 1 , wherein the intracellular domain of the second CAR is selected from the group consisting of SEQ ID NO: 2014, SEQ ID NO: 2015, and SEQ ID NO: 2016. 
     
     
         5 . A method of treating cancer, comprising administering an immune cell to a patient, wherein the immune cell comprises:
 a first CAR comprising an extracellular domain comprising an antigen binding domain, a transmembrane domain, and an intracellular domain; and   a second CAR, wherein the second CAR comprises:   an extracellular domain comprising an antigen binding domain,   a transmembrane domain, and   an intracellular domain, wherein:
 the intracellular domain comprises an Immunoreceptor Tyrosine-based Switch Motif (ITSM), wherein said ITSM is a sequence of amino acids TX 1 YX 2 X 3 X 4  (SEQ ID NO: 2049), wherein
 X 1  is E; 
 X 2  is A or S; 
 X 3  is S or E; and 
 X 4  is V or I; and 
 
   
       the intracellular domain has at least 95% amino acid sequence identity with SEQ ID NO: 2016. 
     
     
         6 . The method of  claim 5 , wherein the cancer is breast cancer, prostate cancer, ovarian cancer, cervical cancer, skin cancer, pancreatic cancer, colorectal cancer, renal cell cancer, liver cancer, brain cancer, lymphoma, leukemia, or lung cancer. 
     
     
         7 . The method of  claim 6 , wherein the cancer is lymphoma. 
     
     
         8 . The method of  claim 6 , wherein the cancer is leukemia. 
     
     
         9 . The method of  claim 5 , wherein the ITSM is selected from the group consisting of TEYASI (SEQ ID NO: 936), TEYSEI (SEQ ID NO: 940), and TEYSEV (SEQ ID NO: 941). 
     
     
         10 . The method of  claim 9 , wherein the antigen binding domain of the second CAR is a single chain variable fragment (scFv). 
     
     
         11 . The method of  claim 10 , wherein the intracellular domain of the second CAR is selected from the group consisting of SEQ ID NO: 2014, SEQ ID NO: 2015, and SEQ ID NO: 2016. 
     
     
         12 . The method of  claim 6 , wherein the ITSM is selected from the group consisting of TEYASI (SEQ ID NO: 936), TEYSEI (SEQ ID NO: 940), and TEYSEV (SEQ ID NO: 941). 
     
     
         13 . The method of  claim 12 , wherein the antigen binding domain of the second CAR is a single chain variable fragment (scFv). 
     
     
         14 . The method of  claim 13 , wherein the intracellular domain of the second CAR is selected from the group consisting of SEQ ID NO: 2014, SEQ ID NO: 2015, and SEQ ID NO: 2016.