Fgf-2 variants having n-terminal deletions and increased receptor selectivity and uses thereof

Abstract

The present invention relates to the design, manufacture and use of fibroblast growth factor (FGF) polypeptides having improved receptor specificity. In particular, the invention relates to isolated FGF2 and FGF4 polypeptides comprising a truncated N-terminus and optionally N-terminal amino acid substitutions. The present invention provides polypeptides, nucleic acids encoding the polypeptides, compositions comprising same and methods for use thereof.

1 . An isolated variant of a fibroblast growth factor (FGF) polypeptide, wherein the FGF is FGF4 comprising an N-terminal deletion; wherein the variant polypeptide retains between 0 and 11 amino acid residues at the N-terminus extending beyond the Leu-Tyr-Cys (LYC) motif of the β1 strand of the core domain; and wherein the variant polypeptide has increased receptor selectivity when compared to the corresponding isolated wild type FGF polypeptide by a gain of activity or loss of activity by at least a factor of two toward at least one receptor subtype but not toward all fibroblast growth factor receptor (FGFR) subtypes. 
     
     
         2 - 13 . (canceled) 
     
     
         14 . The isolated FGF4 variant polypeptide according to  claim 1 , the variant retaining from between 8 to 11 amino acid residues at the N-terminus extending beyond the LYC motif of the β1 strand of the core domain. 
     
     
         15 . The isolated FGF4 variant polypeptide according to  claim 14  selected from a polypeptide having a sequence set forth in any one of SEQ ID NOS: 63-66. 
     
     
         16 - 17 . (cancelled) 
     
     
         18 . A polynucleotide sequence encoding the isolated FGF4 variant polypeptide of  claim 1 . 
     
     
         19 . A polynucleotide according to  claim 18  the polynucleotide having a sequence set forth in any one of SEQ ID NO: 72-74. 
     
     
         20 . A vector comprising a polynucleotide according to any one of  claim 19 . 
     
     
         21 . A host cell comprising a vector according to  claim 20 . 
     
     
         22 . (canceled) 
     
     
         23 . A pharmaceutical composition comprising as an active ingredient the FGF4 variant of  claim 1 , or a polynucleotide encoding the FGF4 variant of  claim 1 , and a pharmaceutically acceptable diluent or carrier. 
     
     
         24 . (canceled) 
     
     
         25 . The pharmaceutical composition according to  claim 23  formulated for administration via intra-articular, intravenous, intramuscular, subcutaneous, intradermal, or intrathecal routes. 
     
     
         26 . The pharmaceutical composition according to  claim 23  formulated for administration to the site of a bone fracture. 
     
     
         27 . The pharmaceutical composition according to  claim 23  formulated for wound healing. 
     
     
         28 . The pharmaceutical composition according to  claim 23  formulated for treatment of coronary and peripheral vascular disease. 
     
     
         29 . (canceled) 
     
     
         30 . The pharmaceutical composition according to  claim 23  for treating skeletal disorders or coronary and peripheral vascular diseases. 
     
     
         31 - 35 . (canceled) 
     
     
         36 . A method of inducing cellular expansion, comprising the steps of:
 a. isolating a population of cells to be expanded; and   b. exposing said cells to an N-terminal truncated polypeptide variant according to  claim 1 .   
     
     
         37 . The method of  claim 36 , wherein the population of cells to be expanded comprises hematopoietic cells. 
     
     
         38 . The method of  claim 36 , wherein the population of cells to be expanded the cells are selected from stem cells or progenitor cells. 
     
     
         39 . The method of  claim 36 , wherein the population of cells to be expanded comprises cells selected from chondrocytes, osteoblasts, hepatocytes, fibroblasts or mesenchymal, endothelial, epithelial, urothelial, endocrine, neuronal, pancreatic, renal and ocular cell types. 
     
     
         40 - 45 . (canceled)