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

Research output: Contribution to journalArticle

9 Citations (Scopus)

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

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Green Fluorescent Proteins
Transfection
Coronary Vessels
Swine
Smooth Muscle Myocytes
Genes
Arteries
Galactosidases
Tunica Intima
Lac Operon
Femoral Artery
Amylases
Serum
Reporter Genes
Fluorescence Microscopy
Genetic Therapy
Fibrinogen
Hyperplasia
Cardiovascular Diseases
Catheters

All Science Journal Classification (ASJC) codes

  • Surgery

Cite this

Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries. / Pankajakshan, Divya; Makinde, Toluwalope O.; Gaurav, Rohit; Del Core, Michael; Hatzoudis, George; Pipinos, Iraklis; Agrawal, Devendra K.

In: Journal of Surgical Research, Vol. 175, No. 1, 01.06.2012, p. 169-175.

Research output: Contribution to journalArticle

Pankajakshan, Divya ; Makinde, Toluwalope O. ; Gaurav, Rohit ; Del Core, Michael ; Hatzoudis, George ; Pipinos, Iraklis ; Agrawal, Devendra K. / Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries. In: Journal of Surgical Research. 2012 ; Vol. 175, No. 1. pp. 169-175.
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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.",
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AB - 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.

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