Hla class i-restricted t cell receptors against mutated ras

Abstract

Disclosed is an isolated or purified T cell receptor (TCR), wherein the TCR has antigenic specificity for a mutated RAS amino acid sequence presented by a human leukocyte antigen (HLA) Class I molecule. Related polypeptides and proteins, as well as related nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions are also provided. Also disclosed are methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal.

1 . An isolated or purified T-cell receptor (TCR) comprising the amino acid sequences of (i) SEQ ID NOs: 1-3, (ii) SEQ ID NOs: 4-6, or (iii) SEQ ID NOs: 1-6. 
     
     
         2 - 6 . (canceled) 
     
     
         7 . The TCR according to  claim 1 , comprising a human variable region. 
     
     
         8 . The TCR according to  claim 1 , comprising:
 (i) the amino acid sequence of SEQ ID NO: 7,   (ii) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 8, or   (iii) both the amino acid sequence of SEQ ID NO: 7 and an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 8.   
     
     
         9 . The TCR of  claim 1 , further comprising:
 (a) an α chain constant region comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 17, wherein:
 (i) X at position 48 of SEQ ID NO: 17 is Thr or Cys; 
 (ii) X at position 112 of SEQ ID NO: 17 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
 (iii) X at position 114 of SEQ ID NO: 17 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and 
 (iv) X at position 115 of SEQ ID NO: 17 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
   (b) a β chain constant region comprising the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Ser or Cys; or   (c) both (a) and (b).   
     
     
         10 . The isolated or purified TCR of  claim 1 , comprising:
 (a) an α chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 21, wherein:
 (i) X at position 185 of SEQ ID NO: 21 is Thr or Cys; 
 (ii) X at position 249 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
 (iii) X at position 251 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and 
 (iv) X at position 252 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
   (b) a β chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 22, wherein X at position 190 of SEQ ID NO: 22 is Ser or Cys; or   (c) both (a) and (b).   
     
     
         11 . An isolated or purified polypeptide comprising a functional portion of the TCR of  claim 1 , wherein the functional portion comprises the amino acid sequences of:
 (a) all of SEQ ID NOs: 1-3,   (b) all of SEQ ID NOs: 4-6, or   (c) all of SEQ ID NOs: 1-6.   
     
     
         12 . The isolated or purified polypeptide according to  claim 11 , wherein the functional portion comprises:
 (i) the amino acid sequence of SEQ ID NO: 7,   (ii) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 8, or   (iii) both the amino acid sequence of SEQ ID NO: 7 and an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 8.   
     
     
         13 . The isolated or purified polypeptide of  claim 11 , wherein the functional portion comprises:
 (a) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 17, wherein:
 (i) X at position 48 of SEQ ID NO: 17 is Thr or Cys; 
 (ii) X at position 112 of SEQ ID NO: 17 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
 (iii) X at position 114 of SEQ ID NO: 17 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and 
 (iv) X at position 115 of SEQ ID NO: 17 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
   (b) the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Ser or Cys; or   (c) both (a) and (b).   
     
     
         14 . The isolated or purified polypeptide of  claim 11 , comprising:
 (a) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 21, wherein:
 (i) X at position 185 of SEQ ID NO: 21 is Thr or Cys; 
 (ii) X at position 249 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
 (iii) X at position 251 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and 
 (iv) X at position 252 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
   (b) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 22, wherein X at position 190 of SEQ ID NO: 22 is Ser or Cys; or   (c) both (a) and (b).   
     
     
         15 . An isolated or purified protein comprising the polypeptide of  claim 11 . 
     
     
         16 . The isolated or purified protein according to  claim 15 , comprising a first polypeptide chain comprising the amino acid sequences of SEQ ID NOs: 1-3 and a second polypeptide chain comprising the amino acid sequences of SEQ ID NOs: 4-6. 
     
     
         17 . The isolated or purified protein according to  claim 15 , comprising a first polypeptide chain comprising the amino acid sequence of SEQ ID NO: 7 and a second polypeptide chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 8. 
     
     
         18 . The isolated or purified protein of  claim 15 , further comprising:
 (a) a first polypeptide chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 17, wherein:
 (i) X at position 48 of SEQ ID NO: 17 is Thr or Cys; 
 (ii) X at position 112 of SEQ ID NO: 17 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
 (iii) X at position 114 of SEQ ID NO: 17 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and 
 (iv) X at position 115 of SEQ ID NO: 17 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
   (b) a second polypeptide chain comprising the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Ser or Cys; or   (c) both (a) and (b).   
     
     
         19 . The isolated or purified protein of  claim 15 , comprising:
 (a) a first polypeptide chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 21, wherein:
 (i) X at position 185 of SEQ ID NO: 21 is Thr or Cys; 
 (ii) X at position 249 of SEQ ID NO: 21 is Ser, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
 (iii) X at position 251 of SEQ ID NO: 21 is Met, Ala, Val, Leu, Ile, Pro, Phe, or Trp; and 
 (iv) X at position 252 of SEQ ID NO: 21 is Gly, Ala, Val, Leu, Ile, Pro, Phe, Met, or Trp; 
   (b) a second polypeptide chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 22, wherein X at position 190 of SEQ ID NO: 22 is Ser or Cys; or   (c) both (a) and (b).   
     
     
         20 .- 23 . (canceled) 
     
     
         24 . A pharmaceutical composition comprising (a) the TCR according to  claim 1  and (b) a pharmaceutically acceptable carrier. 
     
     
         25 . A method of detecting the presence of cancer in a mammal, the method comprising:
 (a) contacting a sample comprising cells of the cancer with the TCR according to  claim 1 , thereby forming a complex; and   (b) detecting the complex,   wherein detection of the complex is indicative of the presence of cancer in the mammal.   
     
     
         26 . (canceled) 
     
     
         27 . The method according to  claim 25 , wherein the cancer expresses a mutated human RAS amino acid sequence, wherein the mutated human RAS amino acid sequence is a mutated human KRAS, a mutated human HRAS, or a mutated human NRAS amino acid sequence. 
     
     
         28 . The method according to  claim 27 , wherein the mutated human RAS amino acid sequence comprises a wild-type human KRAS, a wild-type human HRAS, or a wild-type human NRAS amino acid sequence with a substitution of glycine at position 12, wherein position 12 is defined by reference to the wild-type human KRAS, wild-type human HRAS, or wild-type human NRAS amino acid sequence, respectively. 
     
     
         29 . The method according to  claim 28 , wherein the substitution is a substitution of glycine at position 12 with valine. 
     
     
         30 . The method according to  claim 27 , wherein the mutated human RAS amino acid sequence is a mutated human Kirsten rat sarcoma viral oncogene homolog (KRAS) amino acid sequence. 
     
     
         31 . The method according to  claim 27 , wherein the mutated human RAS amino acid sequence is a mutated human neuroblastoma rat sarcoma viral oncogene homolog (NRAS) amino acid sequence. 
     
     
         32 . The method according to  claim 27 , wherein the mutated human RAS amino acid sequence is a mutated human Harvey rat sarcoma viral oncogene homolog (HRAS) amino acid sequence. 
     
     
         33 . The method according to  claim 25 , wherein the cancer is pancreatic, colorectal, lung, endometrial, ovarian, or prostate cancer. 
     
     
         34 . A method of treating or preventing cancer in a mammal, comprising administering to the mammal the TCR according to  claim 1  in an amount effective to treat or prevent cancer in the mammal. 
     
     
         35 . The TCR according to  claim 1 , wherein the TCR has antigenic specificity for a mutated human RAS amino acid sequence presented by a human leukocyte antigen (HLA) Class I molecule, and
 wherein the mutated human RAS amino acid sequence is a mutated human Kirsten rat sarcoma viral oncogene homolog (KRAS), a mutated human Harvey rat sarcoma viral oncogene homolog (HRAS), or a mutated human Neuroblastoma rat sarcoma viral oncogene homolog (NRAS) amino acid sequence.   
     
     
         36 . The TCR according to  claim 35 , wherein the amino acid sequence of the mutated human RAS amino acid sequence comprises or consists of SEQ ID NOs: 29 or 30. 
     
     
         37 . The TCR according to  claim 35 , wherein the HLA Class I molecule is an HLA-A molecule. 
     
     
         38 . The TCR according to  claim 35 , wherein the HLA Class I molecule is an HLA-A11 molecule. 
     
     
         39 . The TCR according to  claim 35 , wherein the HLA Class I molecule is an HLA-A*11:01 molecule. 
     
     
         40 . The TCR according to  claim 35 , wherein the mutated human RAS amino acid sequence comprises a wild-type human KRAS, a wild-type human HRAS, or a wild-type human NRAS amino acid sequence with a substitution of glycine at position 12, wherein position 12 is defined by reference to the wild-type human KRAS, wild-type human HRAS, or wild-type human NRAS protein, respectively. 
     
     
         41 . The TCR according to  claim 39 , wherein the substitution is a substitution of glycine at position 12 with valine. 
     
     
         42 . The TCR according to  claim 1 , comprising the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, or both of SEQ ID NOs: 7 and 8. 
     
     
         43 . The TCR according to  claim 1 , comprising
 (a) an α chain constant region comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 17, wherein:
 (i) X at position 48 of SEQ ID NO: 17 is Cys; 
 (ii) X at position 112 of SEQ ID NO: 17 is Leu; 
 (iii) X at position 114 of SEQ ID NO: 17 is Ile; and 
 (iv) X at position 115 of SEQ ID NO: 17 is Val; 
   (b) a β chain constant region comprising the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Cys; or   (c) both (a) and (b).   
     
     
         44 . The TCR according to  claim 1 , comprising:
 (a) an α chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 21, wherein:
 (i) X at position 185 of SEQ ID NO: 21 is Cys; 
 (ii) X at position 249 of SEQ ID NO: 21 is Leu; 
 (iii) X at position 251 of SEQ ID NO: 21 is Ile; and 
 (iv) X at position 252 of SEQ ID NO: 21 is Val; 
   (b) a β chain comprising an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 22, wherein X at position 190 of SEQ ID NO: 22 is Cys; or   (c) both (a) and (b).   
     
     
         45 . The isolated or purified polypeptide according to  claim 11 , wherein the functional portion comprises the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8, or both of SEQ ID NOs: 7 and 8. 
     
     
         46 . The isolated or purified polypeptide of  claim 11 , comprising:
 (a) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 17, wherein:
 (i) X at position 48 of SEQ ID NO: 17 is Cys; 
 (ii) X at position 112 of SEQ ID NO: 17 is Leu; 
 (iii) X at position 114 of SEQ ID NO: 17 is Ile; and 
 (iv) X at position 115 of SEQ ID NO: 17 is Val; 
   (b) the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Cys; or   (c) both (a) and (b).   
     
     
         47 . The isolated or purified polypeptide of  claim 11 , comprising:
 (a) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 21, wherein:
 (i) X at position 185 of SEQ ID NO: 21 is Cys; 
 (ii) X at position 249 of SEQ ID NO: 21 is Leu; 
 (iii) X at position 251 of SEQ ID NO: 21 is Ile; and 
 (iv) X at position 252 of SEQ ID NO: 21 is Val; 
   (b) an amino acid sequence at least 99% identical to the amino acid sequence of SEQ ID NO: 22, wherein X at position 190 of SEQ ID NO: 22 is Cys; or   (c) both (a) and (b).   
     
     
         48 . The isolated or purified protein according to  claim 15 , comprising a first polypeptide chain comprising the amino acid sequence of SEQ ID NO: 7 and a second polypeptide chain comprising the amino acid sequence of SEQ ID NO: 8. 
     
     
         49 . The isolated or purified protein of  claim 15 , comprising:
 (a) an amino acid sequence at least 99% identical to a first polypeptide chain comprising the amino acid sequence of SEQ ID NO: 17, wherein:
 (i) X at position 48 of SEQ ID NO: 17 is Cys; 
 (ii) X at position 112 of SEQ ID NO: 17 is Leu; 
 (iii) X at position 114 of SEQ ID NO: 17 is Ile; and 
 (iv) X at position 115 of SEQ ID NO: 17 is Val; 
   (b) a second polypeptide chain comprising the amino acid sequence of SEQ ID NO: 18, wherein X at position 57 of SEQ ID NO: 18 is Cys; or   (c) both (a) and (b).   
     
     
         50 . The isolated or purified protein of  claim 15 , comprising:
 (a) an amino acid sequence at least 99% identical to a first polypeptide chain comprising the amino acid sequence of SEQ ID NO: 21, wherein:
 (i) X at position 185 of SEQ ID NO: 21 is Cys; 
 (ii) X at position 249 of SEQ ID NO: 21 is Leu; 
 (iii) X at position 251 of SEQ ID NO: 21 is Ile; and 
 (iv) X at position 252 of SEQ ID NO: 21 is Val; 
   (b) an amino acid sequence at least 99% identical to a second polypeptide chain comprising the amino acid sequence of SEQ ID NO: 22, wherein X at position 190 of SEQ ID NO: 22 is Cys; or   (c) both (a) and (b).