表达IL-21的重组乙型肝炎病毒（HBV）作为靶向持久性HBV感染的

StephenW

Theranostics. 2020 Apr 25;10(12):5600-5612. doi: 10.7150/thno.44715. eCollection 2020.
Characterization of IL-21-expressing recombinant hepatitis B virus (HBV) as a therapeutic agent targeting persisting HBV infection.
Shen Z1,2, Wu J1, Gao Z2, Wang J1, Zhu H1, Mao R1, Wang X2,3, Zhang J1,2, Xie Y2, Liu J2,3.
Author information

1
    Department of Infectious Diseases, Huashan Hospital, Fudan University, Shanghai 200040, People's Republic of China.
2
    Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, People's Republic of China.
3
    Key Laboratory of Medical Epigenetics and Metabolism, Institutes of Biomedical Sciences, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, People's Republic of China.

Abstract

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.

© The author(s).
KEYWORDS:

HBV clearance; hepatitis B virus; interleukin 21; recombinant HBV vector; superinfection

PMID:
    32373234
PMCID:
    PMC7196313
DOI:
    10.7150/thno.44715

StephenW

治疗学。 2020 Apr 25; 10（12）：5600-5612。 doi：10.7150 / thno.44715。 eCollection 2020。
表达IL-21的重组乙型肝炎病毒（HBV）作为靶向持久性HBV感染的治疗剂的表征。
沉Z1,2，吴J1，高Z2，王J1，朱H1，毛R1，王X2,3，张J1,2，谢Y2，刘J2,3。
作者信息

1个
    复旦大学附属华山医院传染病科，上海200040
2
    复旦大学上海医学院基础医学院，医学分子病毒学重点实验室（MOE / NHC / CAMS），微生物与寄生虫学系，上海200032
3
    复旦大学上海医学院基础医学学院，生物医学科学研究所，医学表观遗传与代谢重点实验室，上海200032

抽象

乙型肝炎病毒（HBV）引起的慢性感染与肝纤维化，肝硬化和肝细胞癌的高风险有关。在HBV持续性小鼠模型中，白介素21（IL-21）已被确定为病毒清除的有效诱导剂。严格的肝移植性使得重组HBV（rHBV）载体成为肝靶向基因递送的理想选择。以前，我们建立了一个称为5c3c的rHBV载体，它本身具有高度复制性，但需要反式提供的HBV包膜蛋白才能产生病毒体。基于5c3c的rHBV病毒粒子能够将受HBV Sp1启动子驱动的货运基因表达传递到感染的肝细胞中。在这项工作中，我们探索了使用5c3c衍生的rHBV进行肝脏特异性IL-21递送作为治疗慢性HBV感染的可行性。方法：构建具有小鼠或人类IL-21基因（分别称为5c3c-mIL-21和5c3c-hIL-21）的5c3c衍生的rHBV复制子，然后在体内和体外测试其rHBV病毒颗粒的产生。在慢性HBV小鼠模型中确定了5c3c-mIL-21的抗HBV作用。此外，使用HBV感染的HepG2 / NTCP细胞和人肝嵌合体小鼠分析了rHBV病毒体的超感染。结果：5c3c-mIL-21和5c3c-hIL-21在体外和体内均具有包膜蛋白时可以有效复制并产生包膜病毒体。在HBV持久性小鼠模型中，从注射的5c3c-mIL-21复制子表达的IL-21诱导了完全的病毒清除。 5c3c-mIL-21和5c3c-hIL-21病毒体可感染HepG2 / NTCP细胞并引起持续的IL-21表达。最重要的是，表达IL-21的rHBV病毒粒子可以在人肝嵌合小鼠中重叠感染HBV感染的HepG2 / NTCP细胞和人肝细胞，并导致持续的IL-21表达和rHBV产生。结论：这些数据表明，表达5c3c的表达IL-21的rHBV作为治疗慢性HBV感染的新方法具有很高的潜力。

©作者。
关键字：

HBV清除率；乙型肝炎病毒;白介素21;重组HBV载体重复感染

PMID：
    32373234
PMCID：
    PMC7196313
DOI：
    10.7150 / thno.44715

StephenW

https://www.thno.org/v10p5600.htm