title: Baculovirus-mediated vascular endothelial growth factor-D(ΔNΔC)  gene transfer induces angiogenesis in rabbit skeletal muscle.
abstract: BACKGROUND
  Occluded arteries and ischemic tissues cannot always be treated by angioplasty, stenting or by-pass-surgery. Under such circumstances, viral gene therapy may be useful in inducing increased blood supply to ischemic area. There is evidence of improved blood flow in ischemic skeletal muscle and myocardium in both animal and human studies using adenoviral vascular endothelial growth factor (VEGF) gene therapy. However, the expression is transient and repeated gene transfers with the same vector are inefficient due to immune responses.
  
  
  METHODS
  Different baculoviral vectors pseudotyped with or without vesicular stomatitis virus glycoprotein (VSV-G) and/or carrying woodchuck hepatitis virus post-transcriptional regulatory element (Wpre) were tested both in vitro and in vivo. VEGF-D(ΔNΔC) was used as therapeutic transgene and lacZ as a control. In vivo efficacy was evaluated as capillary enlargement and transgene expression in New Zealand White (NZW) rabbit skeletal muscle.
  
  
  RESULTS
  A statistically significant capillary enlargement was detected 6 days after gene transfer in transduced areas compared to the control gene transfers with baculovirus and adenovirus encoding β-galactosidase (lacZ). Substantially improved gene transfer efficiency was achieved with a modified baculovirus pseudotyped with VSV-G and carrying Wpre. Dose escalation experiments revealed that either too large volume or too many virus particles caused inflammation and necrosis in the target tissue, whereas 10(9) plaque forming units injected in multiple aliquots resulted in transgene expression with only mild immune reactions.
  
  
  CONCLUSIONS
  We show the first evidence of biologically significant baculoviral gene transfer in skeletal muscle of NZW rabbits using VEGF-D(ΔNΔC) as a therapeutic transgene.
authors.affiliation: Department of Biotechnology and Molecular Medicine, AI Virtanen Institute, University of Eastern Finland, Kuopio, Finland.
authors.forename: Tommi
authors.initials: T
authors.lastname: Heikura
authors.forename: Tiina
authors.initials: T
authors.lastname: Nieminen
authors.forename: Miia M
authors.initials: MM
authors.lastname: Roschier
authors.forename: Henna
authors.initials: H
authors.lastname: Karvinen
authors.forename: Minna U
authors.initials: MU
authors.lastname: Kaikkonen
authors.forename: Anssi J
authors.initials: AJ
authors.lastname: Mähönen
authors.forename: Hanna P
authors.initials: HP
authors.lastname: Lesch
authors.forename: Tuomas T
authors.initials: TT
authors.lastname: Rissanen
authors.forename: Olli H
authors.initials: OH
authors.lastname: Laitinen
authors.forename: Kari J
authors.initials: KJ
authors.lastname: Airenne
authors.forename: Seppo
authors.initials: S
authors.lastname: Ylä-Herttuala
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002196:Capillaries / Q000254:growth & development; D002199:Capillary Permeability; D005260:Female; D018014:Gene Transfer Techniques / Q000009:adverse effects; D056945:Hep G2 Cells; D006801:Humans; D018482:Muscle, Skeletal / Q000098:blood supply* / Q000473:pathology; D018919:Neovascularization, Physiologic / Q000235:genetics*; D010477:Perfusion; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis; D014161:Transduction, Genetic; D016896:Treatment Outcome; D042643:Vascular Endothelial Growth Factor D / Q000235:genetics* / Q000627:therapeutic use*
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1; D020033:Protein Isoforms; D011994:Recombinant Proteins; D042643:Vascular Endothelial Growth Factor D