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: Jung Hwa Lim, Hyo Jung Shin, Kyeong-Su Park, Chan Hee Lee, Cho-Rok Jung, Dong-Soo Im
• Journal: Molecular therapy : the journal of the American Society of Gene Therapy (Mol Ther) Vol. 20 Issue 4 Pg. 778-87 (Apr 2012) ISSN: 1525-0024 [Electronic] United States
• PMID: 22294149 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Vascular Endothelial Growth Factor A
• ubiquitin carrier proteins
• Fibroblast Growth Factor 2
• Ubiquitin-Conjugating Enzymes

Topics

• Adenoviridae
• Animals
• Cell Proliferation
• Cells, Cultured
• Enzyme-Linked Immunosorbent Assay
• Fibroblast Growth Factor 2 (genetics, metabolism)
• Genetic Therapy (methods)
• Hindlimb (pathology)
• Humans
• Immunohistochemistry
• Ischemia (therapy)
• Mice
• Mice, Inbred BALB C
• Neovascularization, Physiologic (genetics, physiology)
• Ubiquitin-Conjugating Enzymes (genetics, metabolism)
• Vascular Endothelial Growth Factor A (genetics, metabolism)