Characterization of IL-21-expressing recombinant hepatitis B virus (HBV) as a therapeutic agent targeting persisting HBV infection
Characterization of IL-21-expressing recombinant hepatitis B virus (HBV) as a therapeutic agent targeting persisting HBV infection
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表达 IL-21 的重组乙型肝炎病毒 (HBV) 作为针对持续性 HBV 感染的治疗剂的表征
DOI：
10.7150/thno.44715
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发表时间：
2020
期刊：
影响因子：
12.4
通讯作者：
Jing Liu
中科院分区：
文献类型：
--
作者：
Zhongliang Shen;Jingwen Wu;Zixiang Gao;Jingyu Wang;Haoxiang Zhu;Richen Mao;Xuanyi Wang;Jiming Zhang;Youhua Xie;Jing Liu
慢性乙型肝炎病毒（HBV）感染与肝纤维化、肝硬化和肝细胞癌的高风险相关。在HBV持久性小鼠模型中，白细胞介素21 （IL-21）已被确定为病毒清除的有效诱导剂。严格的嗜肝性使得重组HBV （rHBV）载体成为肝脏靶向基因传递的理想载体。先前，我们建立了一种称为5c3c的rhv载体，它本身具有高度复制性，但需要反式中提供的HBV包膜蛋白来产生病毒粒子。基于5c3c的rHBV病毒粒子能够将HBV Sp1启动子驱动的货物基因表达传递到感染的肝细胞中。在这项工作中，我们探讨了使用5c3c来源的rHBV进行肝脏特异性递送IL-21治疗慢性HBV感染的可行性。方法：构建含有小鼠或人IL-21基因的5c3c来源的rHBV复制子（分别命名为5c3c-mIL-21和5c3c-hIL-21），并在体外和体内检测rHBV病毒粒子的产生。在慢性HBV小鼠模型中检测5c3c-mIL-21的抗HBV作用。此外，用hbv感染的HepG2/NTCP细胞和人肝嵌合小鼠分析了rHBV病毒粒子的重复感染。结果：5c3c-mIL-21和5c3c-hIL-21在体外和体内均能高效复制并产生包膜病毒粒子。在小鼠HBV持久性模型中，通过注射5c3c-mIL-21复制子表达的IL-21诱导病毒完全清除。5c3c-mIL-21和5c3c-hIL-21病毒粒子可感染HepG2/NTCP细胞并产生IL-21的持续表达。最重要的是，在人肝脏嵌合小鼠体内，表达IL-21的rHBV病毒粒子能够在hbv感染的HepG2/NTCP细胞和人肝细胞中重叠感染，并产生持续的IL-21表达和rHBV的产生。结论：这些数据表明，5c3c来源的表达il -21的rHBV具有很高的潜力，可以作为一种新的治疗慢性HBV感染的药物。
Chronic infection by hepatitis B virus (HBV) is associated with high risks of liver fibrosis, cirrhosis and hepatocellular carcinoma. In mouse models of HBV persistence, interleukin 21 (IL-21) has been identified as a potent inducer of viral clearance. Strict hepatotropism makes recombinant HBV (rHBV) vectors ideal for liver-targeting gene delivery. Previously, we established an rHBV vector termed 5c3c, which is highly replicative by itself, but requires HBV envelope proteins provided in trans to produce virions. 5c3c-based rHBV virions are capable of delivering cargo gene expression driven by HBV Sp1 promoter into infected hepatocytes. In this work, we explore the feasibility of using 5c3c-derived rHBV for liver-specific delivery of IL-21 as treatment of chronic HBV infection. Methods: 5c3c-derived rHBV replicons harboring mouse or human IL-21 genes (termed 5c3c-mIL-21 and 5c3c-hIL-21 respectively) were constructed and then tested for the production of rHBV virions in vitro and in vivo. 5c3c-mIL-21's anti-HBV effects were determined in chronic HBV mouse model. Furthermore, superinfection by rHBV virions was analysed using HBV-infected HepG2/NTCP cells and human liver chimeric mice. Results: 5c3c-mIL-21 and 5c3c-hIL-21 were efficiently replicative and produced enveloped virions when provided with envelope proteins, both in vitro and in vivo. In mouse model of HBV persistence, IL-21 expressed from injected 5c3c-mIL-21 replicon induced complete viral clearance. 5c3c-mIL-21 and 5c3c-hIL-21 virions could infect HepG2/NTCP cells and engender sustained IL-21 expression. Most importantly, IL-21-expressing rHBV virions could superinfect HBV-infected HepG2/NTCP cells and human hepatocytes in human liver chimeric mice, and engender sustained IL-21 expression and rHBV production. Conclusion: These data suggest the high potential of 5c3c-derived IL-21-expressing rHBV as a novel therapeutic against chronic HBV infection.