Exp Mol Med.
2007 Jun;39(3):412-418.
A novel way of therapeutic angiogenesis using an adeno-associated virus-mediated angiogenin gene transfer
- Affiliations
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- 1Department of Biotechnology, Juseong College, Chungbuk 363-794, Korea. krpark@jsc.ac.kr
- 2Department of Biochemistry, Chungbuk National University, Cheongju 361-763, Korea. sichang@cbnu.ac.kr
- 3School of Dentistry, Chonbuk National University, Jeonju 561-756, Korea.
- 4College of Medicine and Institute for Tumor Research, Chungbuk National University, Cheongju 361-763, Korea.
- 5School of Engineering, University of Suwon, Suwon 445-743, Korea.
Abstract
- To develop a novel therapeutic angiogenesis for the treatment of cardiovascular diseases, angiogenin (ANG1) was examined as a potential therapeutic gene. An adeno-associated virus (AAV)-mediated gene delivery system was used to measure the therapeutic efficacy of ANG1. Using a triple co-transfection technique, rAAV-ANG1-GFP, rAAV- VEGF-GFP and rAAV-GFP vectors were produced, which were then used to infect human umbilical vein endothelial cells (HUVECs) in order to evaluate in vitro angiogenic activities. Their protein expressions, tagged with green fluorescent protein (GFP), were monitored by confocal microscopy. The functional activities were measured using wound-healing HUVEC migration assays. The number of migrated cells stimulated by both the expressed ANG1 and the VEGF in rAAV-infected HUVECs increased almost twice the number observed in the expressed GFP control. In vivo angiogenic activities of the expressed ANG1 or VEGF were determined using mouse angiogenesis assays. The angiogenic activities of ANG1 or VEGF expressed in the injected mice were increased by 1.36 and 2.16 times, respectively, compared to those of the expressed GFP control. These results demonstrate that the expressed ANG1 derived from rAAV infection has in vitro and in vivo angiogenic activities and suggest that the rAAV-ANG1 vector is a potential strategy for therapeutic angiogenesis.
MeSH Terms
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Animals
Cell Movement
Cells, Cultured
Dependovirus/*genetics
Endothelial Cells/metabolism/*physiology
*Gene Transfer Techniques
Genetic Vectors
Humans
Male
Mice
Mice, Inbred C57BL
*Neovascularization, Physiologic
Ribonuclease, Pancreatic/biosynthesis/*genetics
Umbilical Veins/cytology
Vascular Endothelial Growth Factor A/biosynthesis
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