Efficient transduction of vascular endothelial cells with recombinant adeno-associated virus serotype 1 and 5 vectors
Efficient transduction of vascular endothelial cells with recombinant adeno-associated virus serotype 1 and 5 vectors
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DOI：
10.1089/hum.2005.16.235
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发表时间：
2005-02-01
影响因子：
4.2
通讯作者：
Agarwal, A
中科院分区：
文献类型：
--
作者：
Chen, SF;Kapturczak, M;Agarwal, A
重组腺相关病毒(RAAV)因其能同时转导分裂细胞和非分裂细胞、诱导有限的免疫应答和长期转基因表达而成为一种有吸引力的基因治疗工具。以前的研究主要利用2型rAAV，并发现血管细胞的转导效率相对较低。本研究的目的是评价rAAV1-5型在人和大鼠主动脉内皮细胞(HAEC和RAEC)中的转导效率。将含有人α(1)-抗胰蛋白酶(Haat)基因的AAV2反向末端重复序列的重组腺相关病毒载体转化为AAV1至5个血清型的病毒衣壳。还研究了真型rAAV2和编码β-半乳糖苷酶或绿色荧光蛋白的5个载体。与其他血清型(p<；0.001)相比，在转导后7天，感染rAAV1的HAEC和RAEC的转导效率最高。有趣的是，在第14天和第21天，转导rAAV5的细胞的表达水平增加到超过rAAV1的水平。唾液酸酶去除唾液酸可完全抑制rAAV1的转导，而肝素则不起作用。这些研究首次证明了rAAV1在内皮细胞中的转导需要唾液酸残基。与rAAV2相比，rAAV1和5血清型载体在体外和体内转导大鼠主动脉节段的血管内皮细胞和血管内皮细胞中均有显著的转基因表达。这些结果表明rAAV1和rAAV5载体在血管靶向基因治疗策略中具有独特的潜力。
Recombinant adeno-associated virus (rAAV) has become an attractive tool for gene therapy because of its ability to transduce both dividing and nondividing cells, elicit a limited immune response, and the capacity for imparting long-term transgene expression. Previous studies have utilized rAAV serotype 2 predominantly and found that transduction of vascular cells is relatively inefficient. The purpose of the present study was to evaluate the transduction efficiency of rAAV serotypes 1 through 5 in human and rat aortic endothelial cells ( HAEC and RAEC). rAAV vectors with AAV2 inverted terminal repeats containing the human alpha(1)-antitrypsin ( hAAT) gene were transcapsidated using helper plasmids to provide viral capsids for the AAV1 through 5 serotypes. True type rAAV2 and 5 vectors encoding beta-galactosidase or green fluorescence protein were also studied. Infection with rAAV1 resulted in the most efficient transduction in both HAEC and RAEC compared to other serotypes ( p < 0.001) at 7 days posttransduction. Interestingly, expression was increased in cells transduced with rAAV5 to levels surpassing rAAV1 by day 14 and 21. Transduction with rAAV1 was completely inhibited by removal of sialic acid with sialidase, while heparin had no effect. These studies are the first demonstration that sialic acid residues are required for rAAV1 transduction in endothelial cells. Transduction of rat aortic segments ex vivo and in vivo demonstrated significant transgene expression in endothelial and smooth muscle cells with rAAV1 and 5 serotype vectors, in comparison to rAAV2. These results suggest the unique potential of rAAV1 and rAAV5-based vectors for vascular-targeted gene-based therapeutic strategies.