Codon-optimized recombinant plasmid, method of stimulating peripheral nerve regeneration, and method of treating nerve damage in humans

Abstract

Provided is a method for treating a peripheral nervous system damage or injury, or for regenerating peripheral nervous system tissue that involves administering to a subject in need thereof a vector that comprises polynucleotide sequences that encode vascular endothelia growth factor (VEGF) and fibroblast growth factor (FGF2) and further a polynucleotide that encodes resistance to kanamycin. A gene-therapeutic structure coding vascular endothelial growth factor (VEGF) and (FGF-2) is also provided. The gene-therapeutic structure can be administered directly to a damaged nerve and paraneural tissues both in intraoperative and post-operative period to stimulate peripheral nerve regeneration. The structure and method significantly advance existing methods for reconstructive treatment for damaged peripheral nerves.

1 . A method for treating a peripheral nervous system damage or injury, or for regenerating peripheral nervous system tissue, the method comprising administering to a subject in need thereof a vector that comprises polynucleotide sequences that encode vascular endothelia growth factor (VEGF) and fibroblast growth factor (FGF2). 
     
     
         2 . The method of  claim 1 , for treating a peripheral nervous system damage or injury. 
     
     
         3 . The method of  claim 1  that is directed to regenerating peripheral nervous system tissue. 
     
     
         4 . The method of  claim 1 , wherein the vector is administered in vivo. 
     
     
         5 . The method of  claim 1 , wherein the vector is administered to a site of the peripheral nervous system damage or injury or to a tissue to be regenerated. 
     
     
         6 . The method of  claim 1 , wherein the vector is administered to a site of the peripheral nervous system damage or injury at a site proximal or distal to the peripheral nervous system damage, or at sites proximal and distal to said damage. 
     
     
         7 . The method of  claim 1 , comprising administering the vector intra-, peri- and/or paraneurally. 
     
     
         8 . The method of  claim 1 , comprising contacting the vector with a neuron or a Schwann cell, astrocyte, microglia and/or neuron. 
     
     
         9 . The method of  claim 1 , wherein the subject has neurotmesis. 
     
     
         10 . The method of  claim 1 , wherein the subject has a diastatic peripheral nerve damage. 
     
     
         11 . The method of  claim 1 , wherein the subject has peripheral nerve damage other than neurotmesis or diastatic peripheral nerve damage. 
     
     
         12 . The method of  claim 1 , wherein the subject is human. 
     
     
         13 . The method of  claim 1 , wherein the vector further comprises a polynucleotide sequence that encodes resistance to kanamycin. 
     
     
         14 . The method of  claim 1 , wherein the vector comprises FGF2 encoding nucleotides at positions 699-1166 and VEGF165 encoding nucleotides at positions 3723-4298 of SEQ ID NO: 1. 
     
     
         15 . The method of  claim 1 , wherein the vector comprises resistance to kanamycin nucleotides at positions 1469-2511 of SEQ ID NO: 1. 
     
     
         16 . The method of  claim 1 , wherein the vector is pBud(Kan)-VEGF-FGF2 (SEQ ID NO: 1). 
     
     
         16 . A vector comprising polynucleotide sequences that encode vascular endothelia growth factor (VEGF), fibroblast growth factor (FGF2), and resistance to kanamycin. 
     
     
         17 . The vector of  claim 16  that comprises FGF2 encoding nucleotides at positions 699-1166 and VEGF165 encoding nucleotides at positions 3723-4298 of SEQ ID NO: 1. 
     
     
         18 . The vector of  claim 16  having SEQ ID NO: 1. 
     
     
         19 . A cell that has been transformed with the vector of  claim 16 .