TY - JOUR
T1 - Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries
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.
UR - http://www.scopus.com/inward/record.url?scp=84860331282&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84860331282&partnerID=8YFLogxK
U2 - 10.1016/j.jss.2011.02.032
DO - 10.1016/j.jss.2011.02.032
M3 - Article
C2 - 21529824
AN - SCOPUS:84860331282
VL - 175
SP - 169
EP - 175
JO - Journal of Surgical Research
JF - Journal of Surgical Research
SN - 0022-4804
IS - 1
ER -