Receptor tyrosine kinase-like orphan receptor 1 (ror1) single chain fv antibody fragment conjugates and methods of use thereof

Abstract

Compositions including an antibody single-chain variable fragment (scFv) conjugate that specifically binds to ROR1 tumor-associated antigen are provided. The anti-ROR1 scFv antibody and conjugates may include a biologically-active molecule. Such conjugates may comprise a chimeric receptor to direct T cells to respond to ROR1 cancer cells, Methods to use the scFV conjugates to target cells expressing ROR1 for therapeutic and diagnostic purposes are also provided.

1 .- 37 . (canceled) 
     
     
         38 . A method of treating a cancer in a subject comprising administering to said subject an antibody single-chain variable fragment (scFv) comprising:
 (a) a light chain variable region that comprises a CDR L1 corresponding to amino acid position 27-32 of SEQ ID NO: 2, a CDR L2 corresponding to amino acid position 50-52 of SEQ ID NO: 2, and a CDR L3 corresponding to amino acid position 89-97 of SEQ ID NO: 2; and   (b) a heavy chain variable region that comprises a CDR H1 corresponding to amino acid position 151-158 of SEQ ID NO: 2, a CDR H2 corresponding to amino acid position 176-182 of SEQ ID NO: 2, and a CDR H3 corresponding to amino acid position 223-233 of SEQ ID NO: 2, wherein said scFv is capable of binding receptor tyrosine kinase-like orphan receptor 1 (ROR1).   
     
     
         39 . The method of  claim 38 , wherein said antibody single-chain variable fragment is a humanized antibody single-chain variable fragment. 
     
     
         40 . The method of  claim 38 , wherein said antibody single-chain variable fragment forms part of a chimeric antigen receptor. 
     
     
         41 . The method of  claim 40 , wherein said chimeric antigen receptor is expressed on a T cell. 
     
     
         42 . The method of  claim 38 , wherein said cancer comprises cancer cells expressing ROR1. 
     
     
         43 . The method of  claim 38 , wherein said cancer comprises a lymphoma or leukemia. 
     
     
         44 . The method of  claim 43 , wherein said lymphoma or leukemia comprises, chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma, marginal cell B-cell lymphoma, or Burkitt's Lymphoma. 
     
     
         45 . The method of  claim 44 , wherein said lymphoma or leukemia comprises chronic lymphocytic leukemia (CLL). 
     
     
         46 . The method of  claim 38 , wherein said cancer comprises breast cancer, renal cancer, or colon cancer. 
     
     
         47 . The method of  claim 38 , wherein said subject is a human subject. 
     
     
         48 . A nucleic acid encoding an antibody single-chain variable fragment (scFv) comprising:
 (a) a light chain variable region that comprises a CDR L1 corresponding to amino acid position 27-32 of SEQ ID NO: 2, a CDR L2 corresponding to amino acid position 50-52 of SEQ ID NO: 2, and a CDR L3 corresponding to amino acid position 89-97 of SEQ ID NO: 2; and   (b) a heavy chain variable region that comprises a CDR H1 corresponding to amino acid position 151-158 of SEQ ID NO: 2, a CDR H2 corresponding to amino acid position 176-182 of SEQ ID NO: 2, and a CDR H3 corresponding to amino acid position 223-233 of SEQ ID NO: 2, wherein said scFv is capable of binding receptor tyrosine kinase-like orphan receptor 1 (ROR1).   
     
     
         49 . An expression vector comprising said nucleic acid of  claim 48 . 
     
     
         50 . The expression vector of  claim 49 , wherein said expression vector is a viral vector. 
     
     
         51 . The expression vector of  claim 50 , wherein said viral vector is a lentivirus. 
     
     
         52 . A method of making a chimeric antigen receptor T cell comprising transducing a T cell with an expression construct comprising a nucleic acid encoding a chimeric antigen receptor, wherein said chimeric antigen receptor comprises a single-chain variable fragment (scFv) comprising:
 (a) a light chain variable region that comprises a CDR L1 corresponding to amino acid position 27-32 of SEQ ID NO: 2, a CDR L2 corresponding to amino acid position 50-52 of SEQ ID NO: 2, and a CDR L3 corresponding to amino acid position 89-97 of SEQ ID NO: 2; and   (b) a heavy chain variable region that comprises a CDR H1 corresponding to amino acid position 151-158 of SEQ ID NO: 2, a CDR H2 corresponding to amino acid position 176-182 of SEQ ID NO: 2, and a CDR H3 corresponding to amino acid position 223-233 of SEQ ID NO: 2, wherein said scFv is capable of binding receptor tyrosine kinase-like orphan receptor 1 (ROR1).   
     
     
         53 . The method of  claim 52 , wherein said expression construct is a viral vector. 
     
     
         54 . The method of  claim 53 , wherein said viral vector is a lentivirus.