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

doi:10.1016/j.jss.2011.02.032

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 journal › Article › peer-review

11 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 language	English (US)
Pages (from-to)	169-175
Number of pages	7
Journal	Journal of Surgical Research
Volume	175
Issue number	1
DOIs	https://doi.org/10.1016/j.jss.2011.02.032
State	Published - Jun 1 2012
Externally published	Yes

Keywords

adeno-associated virus
gene therapy
intimal hyperplasia
restenosis
vascular smooth muscle cells
vector

ASJC Scopus subject areas

Surgery

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10.1016/j.jss.2011.02.032

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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 journal › Article › peer-review

@article{247e2618dc874244b0c1aa7fdf735440,

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

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.",

keywords = "adeno-associated virus, gene therapy, intimal hyperplasia, restenosis, vascular smooth muscle cells, vector",

author = "Divya Pankajakshan and Makinde, {Toluwalope O.} and Rohit Gaurav and {Del Core}, Michael and George Hatzoudis and Iraklis Pipinos and Agrawal, {Devendra K.}",

note = "Funding Information: This work was supported by the NIH grant R01HL104516 to DKA and LB692 State of Nebraska Tobacco Settlement Funds to Creighton University. The authors thank the Gene Therapy Resource Program (GTRP) of the NIH-NHLBI at the University of Pennsylvania Vector Core facility for providing the gene vectors.",

year = "2012",

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doi = "10.1016/j.jss.2011.02.032",

language = "English (US)",

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AU - Pankajakshan, Divya

AU - Makinde, Toluwalope O.

AU - Gaurav, Rohit

AU - Del Core, Michael

AU - Hatzoudis, George

AU - Pipinos, Iraklis

AU - Agrawal, Devendra K.

N1 - Funding Information: This work was supported by the NIH grant R01HL104516 to DKA and LB692 State of Nebraska Tobacco Settlement Funds to Creighton University. The authors thank the Gene Therapy Resource Program (GTRP) of the NIH-NHLBI at the University of Pennsylvania Vector Core facility for providing the gene vectors.

PY - 2012/6/1

Y1 - 2012/6/1

N2 - 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.

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.

KW - adeno-associated virus

KW - gene therapy

KW - intimal hyperplasia

KW - restenosis

KW - vascular smooth muscle cells

KW - vector

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

Abstract

Keywords

ASJC Scopus subject areas

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