title: Transplantation of modified human adipose derived stromal cells expressing VEGF165 results in more efficient angiogenic response in ischemic skeletal muscle.
abstract: BACKGROUND
  Modified cell-based angiogenic therapy has become a promising novel strategy for ischemic heart and limb diseases. Most studies focused on myoblast, endothelial cell progenitors or bone marrow mesenchymal stromal cells transplantation. Yet adipose-derived stromal cells (in contrast to bone marrow) are abundantly available and can be easily harvested during surgery or liposuction. Due to high paracrine activity and availability ADSCs appear to be a preferable cell type for cardiovascular therapy. Still neither genetic modification of human ADSC nor in vivo therapeutic potential of modified ADSC have been thoroughly studied. Presented work is sought to evaluate angiogenic efficacy of modified ADSCs transplantation to ischemic tissue.
  
  
  METHODS
  Human ADSCs were transduced using recombinant adeno-associated virus (rAAV) serotype 2 encoding human VEGF165. The influence of genetic modification on functional properties of ADSCs and their angiogenic potential in animal models were studied.
  
  
  RESULTS
  We obtained AAV-modified ADSC with substantially increased secretion of VEGF (VEGF-ADSCs). Transduced ADSCs retained their adipogenic and osteogenic differentiation capacities and adhesion properties. The level of angiopoetin-1 mRNA was significantly increased in VEGF-ADSC compared to unmodified cells yet expression of FGF-2, HGF and urokinase did not change. Using matrigel implant model in mice it was shown that VEGF-ADSC substantially stimulated implant vascularization with paralleling increase of capillaries and arterioles. In murine hind limb ischemia test we found significant reperfusion and revascularization after intramuscular transplantation of VEGF-ADSC compared to controls with no evidence of angioma formation.
  
  
  CONCLUSIONS
  Transplantation of AAV-VEGF- gene modified hADSC resulted in stronger therapeutic effects in the ischemic skeletal muscle and may be a promising clinical treatment for therapeutic angiogenesis.
authors.affiliation: Laboratory of angiogenesis, Russian Cardiology Research and Production Complex, 3rd Cherepkovskaya 15A, Moscow, 121552, Russia. dr.eshevchenko@gmail.com
authors.forename: Evgeny K
authors.initials: EK
authors.lastname: Shevchenko
authors.forename: Pavel I
authors.initials: PI
authors.lastname: Makarevich
authors.forename: Zoya I
authors.initials: ZI
authors.lastname: Tsokolaeva
authors.forename: Maria A
authors.initials: MA
authors.lastname: Boldyreva
authors.forename: Veronika Yu
authors.initials: VY
authors.lastname: Sysoeva
authors.forename: Vsevolod A
authors.initials: VA
authors.lastname: Tkachuk
authors.forename: Yelena V
authors.initials: YV
authors.lastname: Parfyonova
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D000818:Animals; D002448:Cell Adhesion; D049109:Cell Proliferation; D017690:Cell Transplantation / Q000379:methods*; D003094:Collagen / Q000737:chemistry; D017077:Culture Media, Conditioned / Q000494:pharmacology; D000229:Dependovirus / Q000378:metabolism; D004338:Drug Combinations; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D007511:Ischemia / Q000628:therapy*; D007797:Laminin / Q000737:chemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D018482:Muscle, Skeletal / Q000473:pathology*; D018919:Neovascularization, Physiologic; D011509:Proteoglycans / Q000737:chemistry; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D017077:Culture Media, Conditioned; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C059485:matrigel; D003094:Collagen