Work Lorraine M, Nicklin Stuart A, Brain Nick J R, Dishart Kate L, Von Seggern Dan J, Hallek Michael, Büning Hildegard, Baker Andrew H
Division of Cardiovascular & Medical Sciences, Glasgow Cardiovascular Research Centre, University of Glasgow, Glasgow G11 6NT, UK.
Mol Ther. 2004 Feb;9(2):198-208. doi: 10.1016/j.ymthe.2003.11.006.
The vascular smooth muscle cell (SMC) is integral to the pathogenesis of neointimal formation associated with late vein graft failure, in-stent restenosis, and transplant arteriopathy. Viral vectors transduce SMC with low efficiency and hence, there is a need for improvement. We aimed to enhance the efficiency and selectivity of gene delivery to human SMC. Targeting ligands were identified using phage display on primary human saphenous vein SMC with linear and cyclic libraries. Two linear peptides, EYHHYNK (EYH) and GETRAPL (GET), were incorporated into the HI loop of adenovirus (Ad) fibers and the capsid protein of adeno-associated virus-2 (AAV-2). Exposure of human venous SMC to EYH-modified (but not the GET-modified) Ad vector resulted in a significant increase in transgene expression levels at short, clinically relevant exposure times. Similarly, the EYH-modified AAV vector resulted in enhanced gene transfer to human venous SMC but not endothelial cells in a time- and dose-dependent manner. The EYH-modified AAV vector also enhanced (up to 70-fold) gene delivery to primary human arterial SMC. Hence, incorporation of EYH into Ad and AAV capsids resulted in a significant and selective enhancement in transduction of SMC and has implications for improving local gene delivery to the vasculature.
血管平滑肌细胞（SMC）在与晚期静脉移植物衰竭、支架内再狭窄和移植血管病相关的新生内膜形成的发病机制中不可或缺。病毒载体转导SMC的效率较低，因此需要改进。我们旨在提高基因传递至人SMC的效率和选择性。使用线性和环状文库在原代人隐静脉SMC上进行噬菌体展示来鉴定靶向配体。将两种线性肽，EYHHYNK（EYH）和GETRAPL（GET），分别掺入腺病毒（Ad）纤维的HI环和腺相关病毒2（AAV-2）的衣壳蛋白中。将人静脉SMC暴露于EYH修饰（而非GET修饰）的Ad载体，在短时间、临床相关暴露时间下，转基因表达水平显著增加。同样，EYH修饰的AAV载体以时间和剂量依赖性方式增强了向人静脉SMC而非内皮细胞的基因转移。EYH修饰的AAV载体还将向原代人动脉SMC的基因传递增强了（高达70倍）。因此，将EYH掺入Ad和AAV衣壳中可显著且选择性地增强SMC的转导，对改善向脉管系统的局部基因传递具有重要意义。
