- 作者：Divya Pankajakshan ; Ph.D. ; Toluwalope O. Makinde ; Ph.D. ; Rohit Gaurav ; M.S. ; Michael Del Core ; M.D. ; George Hatzoudis ; M.D. ; Iraklis Pipinos ; M.D. ; Devendra K. Agrawal ; Ph.D. ; dkagr@creighton.edu
- 关键词：adeno-associated virus ; gene therapy ; intimal hyperplasia ; restenosis ; vascular smooth muscle cells ; vector
- 刊名：Journal of Surgical Research
- 出版年：2012
- 期刊代码：299_00224804
- 类别：med
- 出版时间：1 June, 2012
- 卷：175
- 期：1
- 页码：169-175
- 文件大小：1250 K
Background
Gene therapy has attracted attention for its potential to treat several cardiovascular diseases. The use of adeno-associated viral (AAV) vectors to facilitate therapeutic gene transfer to suppress intimal hyperplasia is a promising concept. The objective of this study was to analyze the in vivo transduction of a novel recombinant AAV-2/9 vector with SM22伪 promoter, containing 尾-galactosidase gene (LacZ) or green fluorescent protein (GFP) as reporter genes, to the medial layer smooth muscle cells (SMCs) of swine coronary and peripheral arteries.Methods
The AAV-2/9 vector containing SM22伪 (1 脳 1013 pfu) were administered into carotid/femoral/coronary arteries of domestic swine using irrigating balloon catheter-based gene delivery. Following gene transfer, cryosections of arteries were processed for X-Gal and GFP analysis. Fluorescence microscopy and Western blotting were done to analyze the GFP expression in the SMCs.
Results
LacZ mRNA expression was visualized in the medial layer 7 d after vector administration. The GFP expression was detected at day 7 and lasted for at least 2 mo showing the longer-lasting expression of the AAV-2/9 vector. Control arteries did not show any expression of GFP or LacZ. There was no significant effect of AAV-2/9 viral transduction on serum amylase, fibrinogen, and serum CRP levels.
Conclusion
These finding support the use of AAV-2/9 as a vector to effectively transduce a gene in SMCs of coronary and peripheral arteries without causing inflammation.