Recombinant adeno-associated virus serotype 9 in a mouse model of atherosclerosis: Determination of the optimal expression time in vivo.
Recombinant adeno-associated virus serotype 9 in a mouse model of atherosclerosis: Determination of the optimal expression time in vivo.
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DOI：
10.3892/mmr.2017.6235
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发表时间：
2017-04
影响因子：
3.4
通讯作者：
Yang Y
中科院分区：
文献类型：
--
作者：
Chen Q;Zhai H;Li X;Ma Y;Chen B;Liu F;Lai H;Xie J;He C;Luo J;Gao J;Yang Y
腺相关病毒9（Adeno-associated virus 9，AAV 9）是目前基因治疗的理想载体之一。本研究的目的是确定使用基于重组杆状病毒（rBac）的系统产生的AAV 9载体的基因转染效率和安全性。使用rBac系统产生AAV 9-巨细胞病毒（CMV）-绿色荧光蛋白，并将所得载体颗粒静脉内注射到小鼠中。在注射后14、21、28、35、60、90和120天处死动物。通过荧光成像和蛋白质印迹分析C57/6 B和载脂蛋白E−/−小鼠主动脉血管和主动脉斑块中的GFP表达。使用肝脏和心脏损伤以及肾功能的生物标志物的体内分析以及体外末端脱氧核苷酸转移酶dUTP缺口末端标记分析来确定AAV 9载体的毒性。本研究的结果表明，使用rBac系统包装的AAV 9病毒载体在动脉粥样硬化斑块中适当地起作用。CMV启动子以时间依赖性方式显著诱导血管斑块中的GFP表达。AAV 9-CMV病毒颗粒未导致心脏、肝脏或肾脏损伤，并且未鉴定到细胞凋亡率的变化。这些结果表明，AAV 9-CMV可以有效和安全地用于将基因导入动脉粥样硬化斑块。
Adeno-associated virus 9 (AAV9) has been identified as one of the optimal gene transduction carriers for gene therapy. The aim of the present study was to determine the gene transfection efficiency and safety of an AAV9 vector produced using a recombinant baculovirus (rBac)-based system. AAV9-cytomegalovirus (CMV)-green fluorescent protein was produced using an rBac system and the resulting vector particles were injected intravenously into mice. Animals were sacrificed at 14, 21, 28, 35, 60, 90 and 120 days following injection. GFP expression in aortic vasculature and aortic plaques in C57/6B and apolipoprotein E−/− mice was analyzed by fluorescence imaging and western blotting. In vivo analyses of biological markers of liver and heart damage, and renal function, as well as in vitro terminal deoxynucleotidyl transferase dUTP nick end labeling analysis were used to determine the toxicity of the AAV9 carrier. The findings of the present study demonstrated that AAV9 viral vectors packaged using the rBac system functioned appropriately in arteriosclerosis plaques. The CMV promoter significantly induced GFP expression in the vascular plaque in a time-dependent manner. AAV9-CMV viral particles did not lead to heart, liver or renal damage and no change in apoptotic rate was identified. These findings indicated that AAV9-CMV may be effectively and safely used to transfect genes into atherosclerotic plaques.