Trifunctional t cell-antigen coupler and methods and uses thereof

Abstract

A trifunctional molecule comprising a target-specific ligand, a ligand that binds a protein associated with the TCR complex and a T cell receptor signaling domain polypeptide is provided. Engineering T cells with this novel receptor engenders anti -gen specific activation of numerous T cell functions, including cytokine production, degranulation and cytolysis.

1 . An engineered γδ T Cell comprising an expression vector comprising a nucleic acid encoding a T Cell-Antigen Coupler (TAC) polypeptide, the TAC polypeptide comprising:
 (a) a first antigen-binding domain that binds a tumor antigen; 
 (b) a second antigen-binding domain comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 14 or SEQ ID NO: 25, wherein the second antigen-binding domain binds a CD3 epsilon protein associated with a TCR complex on the engineered γδ T cell; and 
 (c) a T cell co-receptor domain polypeptide comprising a CD4 cytosolic domain and a CD4 transmembrane domain; 
 wherein components (a), (b) and (c) are fused directly to each other or joined by at least one linker, and provided that the TAC polypeptide does not comprise a co-stimulatory domain, and 
 wherein the TAC polypeptide is expressed by the engineered γδ T cell. 
 
     
     
         2 . The engineered γδ T cell of  claim 1 , wherein the first antigen-binding domain binds to human epidermal growth factor receptor 2 (HER2). 
     
     
         3 . The engineered γδ T cell of  claim 1 , wherein the first antigen-binding domain comprises an amino acid sequence as set forth in SEQ ID NO: 8 or SEQ ID NO: 23. 
     
     
         4 . The engineered γδ T cell of  claim 1 , wherein the first antigen-binding domain binds to B-cell maturation antigen (BCMA). 
     
     
         5 . The engineered γδ T cell of  claim 1 , wherein the second antigen-binding domain comprises an amino acid sequence as set forth in SEQ ID NO: 14 or SEQ ID NO: 25. 
     
     
         6 . A pharmaceutical composition comprising the engineered γδ T cell of  claim 1 , and a pharmaceutically acceptable carrier. 
     
     
         7 . A method of providing a cancer immunotherapy for treating a cancer that expresses a tumor antigen in an individual in need thereof, the method comprising administering the engineered γδ T cell of  claim 1  to the individual, wherein the first antigen-binding domain binds to the tumor antigen expressed by the cancer, thereby treating the cancer. 
     
     
         8 . An engineered γδ T cell comprising a T cell Antigen Coupler (TAC) polypeptide, the TAC polypeptide comprising:
 (a) a first antigen-binding domain that binds a tumor antigen; 
 (b) a second antigen-binding domain that binds a CD3 epsilon protein associated with a TCR complex on the engineered γδ T cell; and 
 (c) a T cell co-receptor domain polypeptide comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 18, wherein the T cell co-receptor domain polypeptide comprises a CD4 cytosolic domain and a CD4 transmembrane domain;
 wherein the components (a), (b) and (c) are fused directly to each other or joined by at least one linker, and provided that the TAC polypeptide does not comprise a co-stimulatory domain, and 
 wherein the TAC polypeptide is expressed by the engineered γδ T cell. 
 
 
     
     
         9 . The engineered γδ T cell of  claim 8 , wherein the antigen-binding domain that binds the CD3 epsilon protein associated with the TCR complex on the γδ T cell is UCHT1. 
     
     
         10 . The engineered γδ T cell of  claim 8 , wherein the first antigen-binding domain binds to human epidermal growth factor receptor 2 (HER2). 
     
     
         11 . The engineered γδ T cell of  claim 8 , wherein the first antigen-binding domain comprises an amino acid sequence as set forth in SEQ ID NO: 8 or SEQ ID NO: 23. 
     
     
         12 . The engineered γδ T cell of  claim 8 , wherein the first antigen-binding domain binds to B-cell maturation antigen (BCMA). 
     
     
         13 . A pharmaceutical composition, comprising the engineered γδ T cell of  claim 8 , and a pharmaceutically acceptable carrier. 
     
     
         14 . A method of providing a cancer immunotherapy for treating a cancer that expresses a tumor antigen in an individual in need thereof, the method comprising administering the engineered γδ T cell of  claim 8  to the individual, wherein the first antigen-binding domain binds to the tumor antigen expressed by the cancer, thereby treating the cancer. 
     
     
         15 . An engineered γδ T cell comprising a T cell Antigen Coupler (TAC) polypeptide, the TAC polypeptide comprising:
 (a) a first antigen-binding domain that binds a tumor antigen; 
 (b) a second antigen-binding domain comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 14 or SEQ ID NO: 25, wherein the second antigen-binding domains binds a CD3 epsilon protein associated with a TCR complex on the engineered γδ T cell; and 
 (c) a T cell co-receptor domain polypeptide comprising an amino acid sequence having at least 80% sequence identity with the amino acid sequence as set forth in SEQ ID NO: 18, wherein the T cell co-receptor domain polypeptide comprises a CD4 cytosolic domain and a CD4 transmembrane domain;
 wherein the components (a), (b) and (c) are fused directly to each other or joined by at least one linker, and provided that the nucleic acid does not encode a co-stimulatory domain, and 
 wherein the TAC polypeptide is expressed by the engineered γδ T cell. 
 
 
     
     
         16 . The engineered γδ T cell of  claim 15 , wherein the first antigen-binding domain binds to human epidermal growth factor receptor 2 (HER2). 
     
     
         17 . The engineered γδ T cell of  claim 15 , wherein the first antigen-binding domain comprises the amino acid sequence as set forth in SEQ ID NO: 8 or SEQ ID NO: 23. 
     
     
         18 . The engineered γδ T cell of  claim 15 , wherein the first antigen-binding domain binds to B-cell maturation antigen (BCMA). 
     
     
         19 . A pharmaceutical composition, comprising the engineered γδ T cell of  claim 15 , and a pharmaceutically acceptable carrier. 
     
     
         20 . A method of providing a cancer immunotherapy for treating a cancer that expresses a tumor antigen in an individual in need thereof, the method comprising administering the engineered γδ T cell of  claim 15  to the individual, wherein the first antigen-binding domain binds to the tumor antigen expressed by the cancer, thereby treating the cancer.