title: Activated forms of VEGF-C and VEGF-D provide improved vascular function in skeletal muscle.
abstract: The therapeutic potential of vascular endothelial growth factor (VEGF)-C and VEGF-D in skeletal muscle has been of considerable interest as these factors have both angiogenic and lymphangiogenic activities. Previous studies have mainly used adenoviral gene delivery for short-term expression of VEGF-C and VEGF-D in pig, rabbit, and mouse skeletal muscles. Here we have used the activated mature forms of VEGF-C and VEGF-D expressed via recombinant adeno-associated virus (rAAV), which provides stable, long-lasting transgene expression in various tissues including skeletal muscle. Mouse tibialis anterior muscle was transduced with rAAV encoding human or mouse VEGF-C or VEGF-D. Two weeks later, immunohistochemical analysis showed increased numbers of both blood and lymph vessels, and Doppler ultrasound analysis indicated increased blood vessel perfusion. The lymphatic vessels further increased at the 4-week time point were functional, as shown by FITC-lectin uptake and transport. Furthermore, receptor activation and arteriogenic activity were increased by an alanine substitution mutant of human VEGF-C (C137A) having an increased dimer stability and by a chimeric CAC growth factor that contained the VEGF receptor-binding domain flanked by VEGF-C propeptides, but only the latter promoted significantly more blood vessel perfusion when compared to the other growth factors studied. We conclude that long-term expression of VEGF-C and VEGF-D in skeletal muscle results in the generation of new functional blood and lymphatic vessels. The therapeutic value of intramuscular lymph vessels in draining tissue edema and lymphedema can now be evaluated using this model system.
authors.affiliation: Molecular/Cancer Biology Laboratory, Biomedicum Helsinki, Department of Pathology, Haartman Institute and Helsinki University Central Hospital, University of Helsinki, Finland.
authors.forename: Andrey
authors.initials: A
authors.lastname: Anisimov
authors.forename: Annamari
authors.initials: A
authors.lastname: Alitalo
authors.forename: Petra
authors.initials: P
authors.lastname: Korpisalo
authors.forename: Jarkko
authors.initials: J
authors.lastname: Soronen
authors.forename: Seppo
authors.initials: S
authors.lastname: Kaijalainen
authors.forename: Veli-Matti
authors.initials: VM
authors.lastname: Leppänen
authors.forename: Michael
authors.initials: M
authors.lastname: Jeltsch
authors.forename: Seppo
authors.initials: S
authors.lastname: Ylä-Herttuala
authors.forename: Kari
authors.initials: K
authors.lastname: Alitalo
mesh_terms: D000818:Animals; D001808:Blood Vessels / Q000502:physiology*; D019281:Dimerization; D004355:Drug Stability; D006321:Heart / Q000502:physiology*; D006801:Humans; D042601:Lymphatic Vessels / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000098:blood supply / Q000502:physiology*; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D011994:Recombinant Proteins / Q000378:metabolism; D042561:Vascular Endothelial Growth Factor B / Q000235:genetics / Q000502:physiology*; D042582:Vascular Endothelial Growth Factor C / Q000235:genetics / Q000502:physiology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000502:physiology
chemical_list: D011994:Recombinant Proteins; C467485:VEGFB protein, human; C467486:VEGFC protein, human; D042561:Vascular Endothelial Growth Factor B; D042582:Vascular Endothelial Growth Factor C; C467482:vascular endothelial growth factor B, mouse; C467483:vascular endothelial growth factor C, mouse; D040301:Vascular Endothelial Growth Factor Receptor-2