title: Adenovirus-mediated E2-EPF UCP gene transfer prevents autoamputation in a mouse model of hindlimb ischemia.
abstract: E2-EPF ubiquitin carrier protein (UCP) stabilizes hypoxia-inducible factor-1α (HIF-1α) inducing ischemic vascular responses. Here, we investigated the effect of UCP gene transfer on therapeutic angiogenesis. Adenovirus-encoded UCP (Ad-F-UCP) increased the expression of vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF-2) in cells and mice. Conditioned media from UCP-overexpressing cells promoted proliferation, tubule formation, and invasion of human umbilical-vascular-endothelial cells (HUVECs), and vascularization in chorioallantoic membrane (CAM) assay. Ad-F-UCP increased the vessel density in the Martigel plug assay, and generated copious vessel-like structures in the explanted muscle. The UCP effect on angiogenesis was dependent on VEGF and FGF-2. In mouse hindlimb ischemia model (N = 30/group), autoamputation (limb loss) occurred in 87% and 68% of the mice with saline and Ad encoding β-galactosidase (Ad-LacZ), respectively, whereas only 23% of the mice injected with Ad-F-UCP showed autoamputation after 21 days of treatment. Ad-F-UCP increased protein levels of HIF-1α, platelet-endothelial cell adhesion molecule-1 (PECAM-1), smooth muscle cell actin (SMA) in the ischemic muscle, and augmented blood vessels doubly positive for PECAM-1 and SMA. Consequently, UCP gene transfer prevented muscle degeneration and autoamputation of ischemic limb. The results suggest that E2-EPF UCP may be a target for therapeutic angiogenesis.
authors.affiliation: Gene Therapy Research Unit, KRIBB, Daejeon, Republic of Korea.
authors.forename: Jung Hwa
authors.initials: JH
authors.lastname: Lim
authors.forename: Hyo Jung
authors.initials: HJ
authors.lastname: Shin
authors.forename: Kyeong-Su
authors.initials: KS
authors.lastname: Park
authors.forename: Chan Hee
authors.initials: CH
authors.lastname: Lee
authors.forename: Cho-Rok
authors.initials: CR
authors.lastname: Jung
authors.forename: Dong-Soo
authors.initials: DS
authors.lastname: Im
mesh_terms: D000256:Adenoviridae; D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods; D006614:Hindlimb / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D007511:Ischemia / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D018919:Neovascularization, Physiologic / Q000235:genetics / Q000502:physiology; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
chemical_list: D042461:Vascular Endothelial Growth Factor A; C046288:ubiquitin carrier proteins; D016222:Fibroblast Growth Factor 2; D044763:Ubiquitin-Conjugating Enzymes