Engineered fgf1 and fgf2 compositions and methods of use thereof

Abstract

Engineered FGF1 and FGF2 polypeptides, polynucleotides encoding these polypeptides and DNA constructs, vectors and compositions including these engineered polypeptides are provided herein. The engineered FGF1 and FGF2 polypeptides are more stable than their wild-type counterparts and may be more effective at treating a variety of conditions that FGF1 and FGF2 are useful for treating such as wound healing.

1 . (canceled) 
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . A FGF2 polypeptide comprising at least 80% sequence identity to SEQ ID NO: 3 and engineered to comprise at least one amino acid substitution to SEQ ID NO: 3 selected from the group consisting of Q65L, Q65A, Q65P, Q65V, Q65I, Q65M, Q65F, Q65W, Q65Y, N111S, N111Q, N111T, N111Y, K128N, K128Q, K128H, K128R, K128E, K128D, and K128P, or a functional fragment of the FGF2 polypeptide thereof. 
     
     
         6 . The FGF2 polypeptide of  claim 5 , further comprising a K138E or K138D amino acid substitution to SEQ ID NO: 3. 
     
     
         7 . The FGF2 polypeptide of  claim 6 , wherein the FGF2 polypeptide comprises at least 80% sequence identity to SEQ ID NO: 4 and comprises amino acid residues 65, 111, 128, and 138 of SEQ ID NO: 4, or a functional fragment of the FGF2 polypeptide thereof. 
     
     
         8 . The FGF2 polypeptide of  claim 7 , wherein the FGF2 polypeptide comprises SEQ ID NO: 4. 
     
     
         9 . A fusion protein comprising the composition of  claim 5  and a membrane permeable peptide. 
     
     
         10 . A polynucleotide encoding the polypeptide of  claim 5 . 
     
     
         11 . The polynucleotide of  claim 10 , wherein the polynucleotide is codon-optimized for expression in a cell. 
     
     
         12 . The polynucleotide of  claim 11 , wherein the cell is a prokaryotic cell. 
     
     
         13 . The polynucleotide of  claim 11 , wherein the cell is mammalian cell. 
     
     
         14 . The polynucleotide of  claim 10 , wherein the polynucleotide is DNA. 
     
     
         15 . A DNA construct comprising a promoter operably linked to the polynucleotide of  claim 10 . 
     
     
         16 . The DNA construct of  claim 15 , wherein the promoter is a heterologous promoter. 
     
     
         17 . The DNA construct of  claim 15 , wherein the promoter is a mammalian promoter. 
     
     
         18 . A vector comprising the polynucleotide of  claim 10 . 
     
     
         19 . (canceled) 
     
     
         20 . A pharmaceutical composition comprising the composition of  claim 5  and a pharmaceutical carrier. 
     
     
         21 . A hydrogel comprising the composition of  claim 5 . 
     
     
         22 . A method of treating a condition comprising administering the composition of  claim 5  to a subject in an amount effective to treat the condition, wherein the condition is selected from the group consisting of a wound, Type 1 diabetes, Type 2 diabetes, obesity, internal injuries, a cardiovascular disorder, a cosmetic condition, critical limb ischemia, a nerve injury, a burn, hair loss, a retinal disorder, a muscular disorder, an arterial disease, an age related disorder, organ or tissue damage, osteoporosis, a digestive tract ulcer, ulcerative colitis, a scar, an energy homeostasis disorder, osteoarthritis, and acute renal failure. 
     
     
         23 . The method of  claim 22 , wherein the condition is a wound. 
     
     
         24 . The method of  claim 22 , wherein the subject is a mammal. 
     
     
         25 . The method of  claim 24 , wherein the mammal is a human.