Inhibition of HBV replication and gene expression in vitro and in vivo with a single AAV vector delivering two shRNA molecules
Inhibition of HBV replication and gene expression in vitro and in vivo with a single AAV vector delivering two shRNA molecules
复制标题
使用传递两个 shRNA 分子的单个 AAV 载体在体外和体内抑制 HBV 复制和基因表达
DOI：
10.5483/bmbrep.2009.42.1.059
复制
发表时间：
2009-01-31
期刊：
影响因子：
3.8
通讯作者：
Kung, Hsiang-fu
中科院分区：
文献类型：
--
作者：
Li, Zhi;He, Ming-liang;Kung, Hsiang-fu
乙肝病毒感染在世界范围内高度流行。目前抗病毒治疗的主要挑战是通过快速病毒突变而产生的耐药性增加。在本研究中，我们开发了AAV载体，以同时传递两到三个针对不同乙肝相关基因的shRNA。这些载体在体外表现出明显比单一shRNA载体更好的抗病毒效果。同时表达HBVX基因和S基因的双shRNA表达载体(AAV157I/1694I)可使HBs Ag、HBeAg和HBVDNA水平分别下降87+/-4、80.3+/-2.6和86.2+/-7%。在预防性治疗的小鼠模型中，HBs Ag和HBeAg被降低到无法检测的水平，血清HBVDNA水平至少降低了100倍。这些结果表明，AAV-157I/1694I具有很强的抗乙肝病毒作用，构建多shRNA表达载体的策略可能会增强抗乙肝病毒的效果，克服病毒的逃避机制，从而导致耐药性的产生。[BMB报告2009；42(1)：59-]
Hepatitis B virus (HBV) infection is highly prevalent worldwide. The major challenge for current antiviral treatment is the elevated drug resistance that occurs via rapid viral mutagenesis. In this study, we developed AAV vectors to simultaneously deliver two or three shRNAs targeting different HBV-related genes. These vectors showed markedly better antiviral effects than ones that delivered a single shRNA in vitro. A dual shRNA expression vector (AAV-157i/1694i), which simultaneously expressed two shRNAs targeted the S and X genes of HBV, reduced HBsAg, HBeAg and HBV DNA levels by 87 +/- 4,80.3 +/- 2.6 and 86.2 +/- 7% respectively, eight days post-transduction. In a mouse model of prophylactic treatment HBsAg and HBeAg were reduced to undetectable levels and the serum HBV DNA level was reduced by at least 100 fold. These results indicate that AAV-157i/1694i generates potent anti-HBV effects and that the strategy of constructing multi-shRNA expression vectors may lead to enhanced anti-HBV efficacy and overcome the evading mechanism of the virus and thus the development of drug resistance. [BMB reports 2009; 42(1): 59-64]