Universal platform for car therapy targeting a novel antigenic signature of cancer

Abstract

The present invention provides a method of identifying a target for preparing an inhibitory chimeric antigen receptor (iCAR) or a protective chimeric antigen receptor (pCAR) capable of preventing or attenuating undesired activation of an effector immune cell. Also provided are a list of iCAR targets, as well as vectors and transduced effector immune cells comprising the nucleic acid molecule and methods for treatment of cancer comprising administering the transduced effector immune cells are further provided.

What is claimed: 
     
         1 . A method for treating cancer in a subject having a tumor characterized by loss of heterozygosity (LOH), comprising administering to the subject an effector immune cell, wherein the effector immune cell comprises on its cell surface:
 i) an inhibitory chimeric antigen receptor (iCAR) comprising:
 an extracellular domain comprising a single chain variable fragment (ScFv) that specifically binds to a single allelic variant of a polymorphic cell surface epitope, wherein the allelic variant is absent from tumor cells of a subject due to loss of heterozygosity (LOH) but present at least on all cells of related normal tissue of the subject; 
 an intracellular domain comprising a signal transduction element that is able to inhibit to effector immune cell, wherein the signal transduction element is homologous to a signal transduction element of PD1, CTLA4, or LIR1; and 
 a hinge domain and a transmembrane domain linking the extracellular domain to the intracellular domain; and 
   ii) an activating chimeric antigen receptor (aCAR) comprising:
 an extracellular domain that specifically binds to another cell surface antigen, wherein the another cell surface antigen is a tumor-associated antigen or is shared at least by cells of related tumor and normal tissue; 
 an intracellular domain comprising at least one signal transduction element that activates and/or co-stimulates an effector immune cell; and 
 a hinge domain and a transmembrane domain linking the extracellular domain of the aCAR construct to the intracellular domain of the aCAR construct. 
   
     
     
         2 . The method of  claim 1 , wherein the signal transduction element is homologous to a signal transduction element of LIR1. 
     
     
         3 . The method of  claim 1 , wherein the tumor is a solid tumor. 
     
     
         4 . The method of  claim 1 , wherein the another cell surface antigen is selected from the group consisting of CD19, CD20, CD22, Igκ, ROR1, CD30, CD174, CD33, CD123, NKG2D-L, CD139, BCMA, GD2, FR-α, L1-CAM, ErbB2, EGFRvIII, VEGFR-2, IL-13Rα2, FAP, Mesothelin, c-MET, PSMA, CEA, EGFR, 5T4, GPC3,  CD38 , CS1, PSCA, CD44v6, CD44v7/8, MUC1, MUC16, PD-L1, IL-11Rα, EphA2, CAIX, and CSPG4. 
     
     
         5 . A nucleic acid molecule encoding an inhibitory chimeric antigen receptor (iCAR) construct, the iCAR construct comprising:
 i) an extracellular domain comprising a single chain variable fragment (ScFv) that specifically binds to a single allelic variant of a polymorphic cell surface epitope, wherein the allelic variant is absent from tumor cells of a subject due to loss of heterozygosity (LOH) but present at least on all cells of related normal tissue of the subject;   ii) an intracellular domain comprising a signal transduction element, wherein the signal transduction element is homologous to a signal transduction element of PD1, CTLA4, or LIR1; and   iii) a hinge domain and a transmembrane domain linking the extracellular domain to the intracellular domain.   
     
     
         6 . The nucleic acid molecule of  claim 5 , wherein the signal transduction element is homologous to a signal transduction element of LIR1. 
     
     
         7 . The nucleic acid molecule of  claim 5 , wherein the tumor is a solid tumor. 
     
     
         8 . A vector comprising the nucleotide sequence of the nucleic acid molecule of  claim 5 , wherein at least one control element, such as a promoter, is operably linked to the nucleotide sequence. 
     
     
         9 . The vector of  claim 8 , further comprising a nucleotide sequence encoding an activating chimeric antigen receptor (aCAR) construct, wherein the aCAR construct comprises:
 i) an extracellular domain that specifically binds to another cell surface antigen, wherein the another cell surface antigen is a tumor-associated antigen or is shared at least by cells of related tumor and normal tissue;   ii) an intracellular domain comprising at least one signal transduction element that activates and/or co-stimulates an effector immune cell; and   iii) a hinge domain and a transmembrane domain linking the extracellular domain of the aCAR construct to the intracellular domain of the aCAR construct.   
     
     
         10 . The vector of  claim 9 , wherein the another cell surface antigen is selected from the group consisting of CD19, CD20, CD22, Igκ, ROR1, CD30, CD174, CD33, CD123, NKG2D-L, CD139, BCMA, GD2, FR-α, L1-CAM, ErbB2, EGFRvIII, VEGFR-2, IL-13Rα2, FAP, Mesothelin, c-MET, PSMA, CEA, EGFR, 5T4, GPC3,  CD38 , CS1, PSCA, CD44v6, CD44v7/8, MUC1, MUC16, PD-L1, IL-11Rα, EphA2, CAIX, and CSPG4. 
     
     
         11 . An effector immune cell comprising on its cell surface an inhibitory chimeric antigen receptor (iCAR) construct comprising:
 i) an extracellular domain comprising a single chain variable fragment (ScFv) that specifically binds to a single allelic variant of a polymorphic cell surface epitope, wherein the allelic variant is absent from tumor cells of a subject due to loss of heterozygosity (LOH) but present at least on all cells of related normal tissue of the subject;   ii) an intracellular domain comprising a signal transduction element, wherein the signal transduction element is homologous to a signal transduction element of PD1, CTLA4, or LIR1; and   iii) a hinge domain and a transmembrane domain linking the extracellular domain to the intracellular domain.   
     
     
         12 . The effector immune cell of  claim 11 , wherein the signal transduction element is homologous to a signal transduction element of LIR1. 
     
     
         13 . The effector immune cell of  claim 11 , wherein the tumor is a solid tumor. 
     
     
         14 . The effector immune cell of  claim 11 , the cell further comprising on its cell surface an aCAR construct comprising:
 i) an extracellular domain that specifically binds to another cell surface antigen, wherein the another cell surface antigen is a tumor-associated antigen or is shared at least by cells of related tumor and normal tissue;   ii) an intracellular domain comprising at least one signal transduction element that activates and/or co-stimulates an effector immune cell; and   iii) a hinge domain and a transmembrane domain linking the extracellular domain of the aCAR construct to the intracellular domain of the aCAR construct.   
     
     
         15 . The effector immune cell of  claim 14 , wherein the another cell surface antigen is selected from the group consisting of CD19, CD20, CD22, Igκ, ROR1, CD30, CD174, CD33, CD123, NKG2D-L, CD139, BCMA, GD2, FR-α, L1-CAM, ErbB2, EGFRvIII, VEGFR-2, IL-13Rα2, FAP, Mesothelin, c-MET, PSMA, CEA, EGFR, 5T4, GPC3,  CD38 , CS1, PSCA, CD44v6, CD44v7/8, MUC1, MUC16, PD-L1, IL-11Rα, EphA2, CAIX, and CSPG4.