Mokhashi Nikita, Choi Robert Y, Cicalese Stephanie, Eguchi Kunie, Boyer Michael J, Cooper Hannah A, Kimura Yayoi, Akiyama Tomoko, Scalia Rosario, Rizzo Victor, Eguchi Satoru
Cardiovascular Research Center, Lewis Katz School of Medicine, Temple University, Philadelphia, PA; and.
Advanced Medical Research Center, Yokohama City University, Yokohama, Japan.
J Cardiovasc Pharmacol. 2020 Jun;75(6):603-607. doi: 10.1097/FJC.0000000000000821.
Adenoviral vectors are useful tools in manipulating a gene of interest in vitro and in vivo, including in the vascular system. The transduction efficiencies of adenoviral vectors in vascular cells such as endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) are known to be lower than those in epithelial cell types. The effective entry for adenoviral vectors is primarily mediated through the coxsackievirus and adenovirus receptor (CAR), which has been shown to be expressed in both cell types. Cationic liposomes have been used to enhance adenovirus transduction efficiency in nonepithelial cells. Accordingly, the aim of this study is to obtain new information regarding differences in transduction efficiencies, cationic liposome sensitivity, and CAR expression between ECs and VSMCs. Using cultured rat aortic ECs and VSMCs, here, we have compared transduction efficiency of adenoviruses with or without inclusion of liposomes and CAR expression. A significant increase in basal transduction efficiency was observed in ECs compared with VSMCs. Cationic liposome polybrene enhanced transduction efficiency in VSMCs, whereas decreased efficiency was observed in ECs. Western blotting demonstrated expression of the CAR in ECs but not in VSMCs. Proteomic analysis and mouse aorta immunostaining further suggests significant expression of the CAR in ECs but not in VSMCs. In conclusion, adenoviruses can effectively transduce the gene of interest in aortic ECs likely because of abundant expression of the CAR, whereas cationic liposomes such as polybrene enhance the transduction efficiency in VSMCs lacking CAR expression.
腺病毒载体是在体外和体内（包括在血管系统中）操纵感兴趣基因的有用工具。已知腺病毒载体在内皮细胞（ECs）和血管平滑肌细胞（VSMCs）等血管细胞中的转导效率低于上皮细胞类型。腺病毒载体的有效进入主要通过柯萨奇病毒和腺病毒受体（CAR）介导，已证明这两种细胞类型中均有表达。阳离子脂质体已被用于提高腺病毒在非上皮细胞中的转导效率。因此，本研究的目的是获取有关ECs和VSMCs之间转导效率、阳离子脂质体敏感性和CAR表达差异的新信息。在这里，我们使用培养的大鼠主动脉ECs和VSMCs，比较了含或不含脂质体的腺病毒的转导效率和CAR表达。与VSMCs相比，ECs的基础转导效率显著增加。阳离子脂质体聚凝胺提高了VSMCs的转导效率，而在ECs中观察到效率降低。蛋白质印迹法显示CAR在ECs中表达，但在VSMCs中不表达。蛋白质组学分析和小鼠主动脉免疫染色进一步表明CAR在ECs中大量表达，但在VSMCs中不表达。总之，腺病毒可能由于CAR的丰富表达而能有效地转导主动脉ECs中感兴趣的基因，而阳离子脂质体如聚凝胺则提高了缺乏CAR表达的VSMCs的转导效率。
