Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries.
Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries.
复制标题
使用 AAV-2/9 载体在猪冠状动脉和外周动脉中成功转染基因。
DOI：
10.1016/j.jss.2011.02.032
复制
发表时间：
2012-06-01
期刊：
影响因子：
--
通讯作者：
Agrawal DK
中科院分区：
文献类型：
--
作者：
Pankajakshan D;Makinde TO;Gaurav R;Del Core M;Hatzoudis G;Pipinos I;Agrawal DK
基因治疗因其治疗多种心血管疾病的潜力而引起人们的关注。使用腺相关病毒（AAV）载体促进治疗性基因转移以抑制内膜增生是一个很有前途的概念。本研究的目的是分析一种新型的重组腺相关病毒2/9载体（含SM 22 α启动子，并含有β-半乳糖苷酶基因（Lac Z）或绿色荧光蛋白（GFP）作为报告基因）对猪冠状动脉和外周动脉中层平滑肌细胞（SMC）的体内转导作用。将含有SM 22 α的AAV 2/9载体（1×1013 pfu）通过基于灌注球囊导管的基因递送方式施用到家猪的颈动脉/股动脉/冠状动脉中。基因转移后，处理动脉的冷冻切片用于X-Gal和GFP分析。采用荧光显微镜和Western blotting检测SMC中GFP的表达。载体给药后7天，在中层中可见LacZ mRNA表达。在第7天检测到GFP表达，并且持续至少2个月，表明AAV 2/9载体的持续表达时间更长。对照动脉没有显示任何GFP或LacZ的表达。AAV 2/9病毒转导对血清淀粉酶、纤维蛋白原和血清CRP水平没有显著影响。这些发现支持使用AAV 2/9作为载体，以有效地将a基因转染到冠状动脉和外周动脉的SMC中，而不引起炎症。
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 (Lac Z) or green fluorescent protein (GFP) as reporter genes, to the medial layer smooth muscle cells (SMCs) of swine coronary and peripheral arteries. The AAV2/9 vector containing SM22α (1×1013 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. LacZ mRNA expression was visualized in the medial layer 7 days after vector administration. The GFP expression was detected at 7th day and lasted for at least 2 months showing the longer-lasting expression of the AAV2/9-vector. Control arteries did not show any expression of GFP or LacZ. There was no significant effect of AAV2/9 viral transduction on serum amylase, fibrinogen and serum CRP levels. These finding support the use of AAV2/9 as a vector to effectively transduce a gene in SMCs of coronary and peripheral arteries without causing inflammation.
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DOI：
10.1073/pnas.94.4.1426
发表时间：
1997-02-18
影响因子：
11.1
作者：
Koeberl, DD;Alexander, IE;Miller, AD
通讯作者：
Miller, AD
DOI：
10.1073/pnas.89.3.1138
发表时间：
1992-02-01
影响因子：
11.1
作者：
LYNCH, CM;CLOWES, MM;MILLER, AD
通讯作者：
MILLER, AD
影响因子：
5.7
作者：
Akyürek, LM;Yang, ZY;Nabel, EG
通讯作者：
Nabel, EG
影响因子：
5.4
作者：
Zaiss, AK;Liu, Q;Muruve, DA
通讯作者：
Muruve, DA
设计师基因递送向量：腺相关病毒载体的分子工程和进化，以增强基因转移。
影响因子：
3.7
作者：
Kwon, Inchan;Schaffer, David V.
通讯作者：
Schaffer, David V.