Antibody/t-cell receptor chimeric constructs and uses thereof

Abstract

The present application provides antibody-TCR chimeric constructs comprising an antibody moiety that specifically binds to a target antigen fused to a TCRM capable of recruiting at least one TCR-associated signaling module. Also provided are methods of making and using these constructs.

What is claimed is: 
     
         1 . An antibody-T cell receptor (TCR) chimeric molecule (abTCR) that specifically binds to a complex comprising an alpha-fetoprotein (AFP) peptide and a major histocompatibility complex (MHC) class I protein (an AFP/MHC class I complex, or AMC), comprising:
 a) a first polypeptide chain, wherein the first polypeptide chain comprises i) a first antigen-binding domain comprising a V H  antibody domain, and ii) a first TCR domain (TCRD) comprising a first transmembrane domain of a first TCR subunit; and   b) a second polypeptide chain, wherein the second polypeptide chain comprises i) a second antigen-binding domain comprising a V L  antibody domain, and ii) a second TCRD comprising a second transmembrane domain of a second TCR subunit,
 wherein the V H  antibody domain and the V L  antibody domain form an antigen-binding module that specifically binds the AMC, 
 wherein the first TCRD and the second TCRD form a TCR module (TCRM) that is capable of recruiting at least one TCR-associated signaling module. 
   
     
     
         2 . The abTCR of  claim 1 , wherein:
 i) the first polypeptide chain further comprises a first peptide linker between the first antigen-binding domain and the first TCRD; and/or   ii) the second polypeptide chain further comprises a second peptide linker between the second antigen-binding domain and the second TCRD.   
     
     
         3 . The abTCR of  claim 2 , wherein the first and/or second peptide linkers comprise, individually, a constant domain or fragment thereof from an immunoglobulin or a TCR subunit. 
     
     
         4 . The abTCR of  claim 5 , wherein the first and/or second peptide linkers comprise, individually, a CH1, CH2, CH3, CH4, or CL antibody domain or fragment thereof. 
     
     
         5 . The abTCR of  claim 5 , wherein the first linker comprises a CH1 antibody domain or fragment thereof, and the second linker comprises a CL antibody domain or fragment thereof. 
     
     
         6 . The abTCR of  claim 5 , wherein the first and/or second peptide linkers comprise, individually, a Cα, Cβ, Cγ, or Cδ TCR domain or fragment thereof. The abTCR of  claim 1 , wherein:
 i) the first TCRD further comprises a first connecting peptide or fragment thereof of a TCR subunit N-terminal to the first transmembrane domain; and/or 
 ii) the second TCRD further comprises a second connecting peptide or fragment thereof of a TCR subunit N-terminal to the second transmembrane domain. 
 
     
     
         8 . The abTCR of  claim 1 , wherein:
 i) the first TCRD further comprises a first TCR intracellular domain comprising a TCR intracellular sequence C-terminal to the first transmembrane domain; and/or   ii) the second TCRD further comprises a second TCR intracellular domain comprising a TCR intracellular sequence C-terminal to the second transmembrane domain.   
     
     
         9 . The abTCR of  claim 1 , wherein:
 i) the first polypeptide chain further comprises a first signaling peptide N-terminal to the first antigen-binding domain; and/or   ii) the second polypeptide chain further comprises a second signaling peptide N-terminal to the second antigen-binding domain.   
     
     
         10 . The abTCR of  claim 1 , wherein:
 i) the first polypeptide chain further comprises a first accessory intracellular domain comprising a co-stimulatory intracellular signaling sequence C-terminal to the first transmembrane domain; and/or   ii) the second polypeptide chain further comprises a second accessory intracellular domain comprising a co-stimulatory intracellular signaling sequence C-terminal to the second transmembrane domain.   
     
     
         11 . The abTCR of  claim 1 , wherein the TCR-associated signaling module is selected from the group consisting of CD3δε, CD3γε, and ζζ. 
     
     
         12 . The abTCR of  claim 1 , wherein:
 i) the first TCR subunit is a TCR α chain, and the second TCR subunit is a TCR β chain; or   ii) the first TCR subunit is a TCR β chain, and the second TCR subunit is a TCR α chain.   
     
     
         13 . The abTCR of  claim 1 , wherein:
 i) the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain; or   ii) the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain.   
     
     
         14 . The abTCR of  claim 1 , wherein:
 i) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 23, and the second polypeptide comprises the amino acid sequence of SEQ ID NO:24;   ii) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 25, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 26;   iii) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 27, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 28;   iv) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 29, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 30;   v) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 31, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 32;   vi) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 33, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 34; or   vii) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 35, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 36.   
     
     
         15 . The abTCR of  claim 1 , wherein the AFP peptide comprises the amino acid sequence of SEQ ID NO: 53. 
     
     
         16 . The abTCR of  claim 1 , wherein the MHC class I protein is the HLA-A*02:01 subtype of HLA-A*02. 
     
     
         17 . The abTCR of  claim 1 , wherein the V H  antibody domain comprises heavy chain complementarity determining region (HC-CDR)1, HC-CDR2 and HC-CDR3 of the amino acid sequence of SEQ ID NO: 38 and the V L  antibody domain comprises light chain complementarity determining region (LC-CDR)1, LC-CDR2 and LC-CDR3 of the amino acid sequence of SEQ ID NO: 40. 
     
     
         18 . Nucleic acid(s) encoding the first and second polypeptide chains of the abTCR of  claim 1 . 
     
     
         19 . An effector cell presenting on its surface the abTCR of  claim 1 . 
     
     
         20 . The effector cell of  claim 19 , wherein the effector cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell. 
     
     
         21 . The effector cell of  claim 19 , wherein the effector cell:
 i) does not express the first TCR subunit and/or the second TCR subunit; or   ii) is modified to block or decrease endogenous expression of the first TCR subunit and/or the second TCR subunit.   
     
     
         22 . A method of killing a target cell presenting an AFP peptide, comprising contacting the target cell with the effector cell of  claim 18 , wherein the abTCR specifically binds to the AFP peptide. 
     
     
         23 . A pharmaceutical composition comprising the effector cell of  claim 19 , and a pharmaceutically acceptable carrier. 
     
     
         24 . A method of treating an AFP-associated disease in an individual in need thereof, comprising administering to the individual an effective amount of the pharmaceutical composition of  claim 23 . 
     
     
         25 . The method of  claim 24 , wherein the AFP-associated disease is a cancer. 
     
     
         26 . The method of  claim 25 , wherein the cancer comprises a solid tumor. 
     
     
         27 . The method of  claim 25 , wherein the cancer is hepatocellular carcinoma.