Repertoire of allo-restricted peptide-specific t cell receptor sequences and use thereof

Abstract

The present invention is directed to a kit-of-parts or composition containing nucleic acid sequences coding for high-avidity, allo-restricted TCR, wherein the TCR are independently directed against the tyrosinase antigen, the melan-A antigen and the survivin antigen. The invention is further directed to a kit-of-parts or composition containing at least three groups of transgenic lymphocytes transformed with vectors coding for TCR against said antigens. Furthermore, the present invention provides a pharmaceutical composition and its use in the treatment of diseases involving malignant cells expressing said tumor-associated antigens. The invention further relates to a nucleic acid molecule coding for a TCR that recognizes the survivin antigen, a TCR encoded thereby and a T cell expressing said TCR. Further, the invention discloses a vector, a cell and a pharmaceutical composition encoding/containing same and their use in the treatment of diseases involving malignant cells expressing survivin.

1 . A nucleic acid molecule coding for the V(D)J regions of a T cell receptor (TCR) that recognizes the survivin antigen and comprising the nucleic acid sequence of SEQ ID NO: 83, 79, 81, or 77 coding for the α-chain and/or the nucleic acid sequence of SEQ ID NO: 84, 80, 82, or 78 coding for the 1-chain of said TCR, or
 a derivative thereof, coding for the α- or β-chain, wherein the chain has been altered by one or more additions or deletions of from 1-15 amino acids, the additions or deletions being outside the CDR3 region of each chain and/or by conservative substitutions of from 1-15 amino acids, wherein the survivin antigen recognizing characteristics are maintained or improved, or 
 a fragment thereof coding for a CDR3 region of a TCR recognizing the survivin antigen and having the nucleic acid sequence of SEQ ID NO: 27, 28, 23, 24, 25, 26, 21, 22 or coding for the amino acid sequences of SEQ ID NO: 55, 56, 51, 52, 53, 54, 49, 50, or 
 a derivative of said fragment, wherein the CDR3 region has been altered by one or more additions and/or deletions of an overall number of from 1-5 amino acids. but not more than 1-3 contiguous amino acids and/or conservative substitutions of from 1-6 amino acids and wherein the survivin antigen recognizing characteristics are maintained or improved. 
 
     
     
         2 . A TCR encoded by a nucleic acid of  claim 1  or comprising one or more of the amino acid sequences of SEQ ID NO: 55, 56, 51, 53, 54, 49, 50. 
     
     
         3 . A functional TCR α and/or β chain fusion protein, comprising:
 a) at least one epitope-tag, and 
 b) the amino acid sequence of an α and/or β chain of a TCR according to  claim 2 , 
 
       wherein said epitope-tag is selected from
 i) an epitope-tag added to the N- and/or C-terminus of said α and/or β chain, or added into the α and/or β chain sequence, but outside the CDR3 region, 
 ii) an epitope-tag inserted into a constant region of said α and/or β chain, and 
 iii) an epitope-tag replacing a number of amino acids in a constant region of said α and/or β chain. 
 
     
     
         4 . A T cell expressing a TCR of  claim 2 . 
     
     
         5 . An immunoglobulin molecule, anticaline, TCR γ/δ chain having a CDR3 region of  claim 1  inserted. 
     
     
         6 . A vector, which comprises one or more of the nucleic acids of  claim 1 . 
     
     
         7 . A cell which has been transformed with the vector of  claim 6 . 
     
     
         8 . A pharmaceutical composition which comprises a TCR of  claim 2  and a pharmaceutically acceptable carrier. 
     
     
         9 . A method of performing adoptive cell therapy, the method comprising administering the pharmaceutical composition of  claim 8  to a patient in need thereof. 
     
     
         10 . A method of treating a disease involving malignant cells expressing survivin in a patient in need thereof, the method comprising administering the pharmaceutical composition of  claim 8  to the patient, thereby treating the disease. 
     
     
         11 . A kit-of-parts or composition comprising:
 a) a group of vectors containing nucleic acid sequences coding for high-avidity, allo-restricted TCR, wherein the TCR are directed against the tyrosinase antigen;   b) a group of vectors containing nucleic acid sequences coding for high-avidity, allo-restricted TCR, wherein the TCR are directed against the melan-A antigen; and   c) a group of vectors containing nucleic acid sequences coding for high-avidity, allo-restricted TCR. wherein the TCR are directed against the survivin antigen.   
     
     
         12 - 22 . (canceled) 
     
     
         23 . A pharmaceutical composition which comprises the kit-of-parts or composition of  claim 11  and a pharmaceutically acceptable carrier, preferably an infusion or injection. 
     
     
         24 - 25 . (canceled) 
     
     
         26 . The TCR of  claim 2 , wherein the TCR is a soluble TCR. 
     
     
         27 . The vector of  claim 6 , wherein the vector is a plasmid, shuttle vector, phagemid, cosmid, expression vector, retroviral vector, adenoviral vector or particle and/or vector to be used in gene therapy. 
     
     
         28 . The cell of  claim 7 , wherein the cell is a peripheral blood lymphocyte (PBL). 
     
     
         29 . The pharmaceutical composition of  claim 8 , wherein the composition is formulated for administration by infusion or injection. 
     
     
         30 . A pharmaceutical composition which comprises a T cell of  claim 4  and a pharmaceutically acceptable carrier. 
     
     
         31 . A method of performing adoptive cell therapy, the method comprising administering the pharmaceutical composition of  claim 30  to a patient in need thereof. 
     
     
         32 . A method of treating a disease involving malignant cells expressing survivin in a patient in need thereof, the method comprising administering the pharmaceutical composition of  claim 30  to the patient, thereby treating the disease.