Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries

Divya Pankajakshan, Toluwalope O. Makinde, Rohit Gaurav, Michael Del Core, George Hatzoudis, Iraklis Pipinos, Devendra K. Agrawal

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10 Scopus citations

Abstract

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 SM22α promoter, containing β-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 SM22α (1 × 10 13 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.

Original languageEnglish (US)
Pages (from-to)169-175
Number of pages7
JournalJournal of Surgical Research
Volume175
Issue number1
DOIs
StatePublished - Jun 1 2012

All Science Journal Classification (ASJC) codes

  • Surgery

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    Pankajakshan, D., Makinde, T. O., Gaurav, R., Del Core, M., Hatzoudis, G., Pipinos, I., & Agrawal, D. K. (2012). Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries. Journal of Surgical Research, 175(1), 169-175. https://doi.org/10.1016/j.jss.2011.02.032