优化的HepaRG是产生用于慢性乙型肝炎病毒感染的人肝脏嵌合

StephenW

Emerg Microbes Infect. 2018 Aug 10;7(1):144. doi: 10.1038/s41426-018-0143-9.
Optimized HepaRG is a suitable cell source to generate the human liver chimeric mouse model for the chronic hepatitis B virus infection.
Yuan L1, Liu X1, Zhang L1, Zhang Y1, Chen Y1, Li X1, Wu K1, Cao J1, Hou W1, Que Y1, Zhang J1, Zhu H2, Yuan Q3, Tang Q4, Cheng T5, Xia N1.
Author information

1
    State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Science, School of Public Health, Xiamen University, 361102, Xiamen, P. R. China.
2
    Department of Microbiology, Biochemistry and Molecular Genetics, New Jersey Medical School, Rutgers University, 225 Warren Street, Newark, NJ, 070101, USA.
3
    State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Science, School of Public Health, Xiamen University, 361102, Xiamen, P. R. China. [email protected].
4
    Department of Microbiology, Howard University College of Medicine, Washington, DC, 20059, USA. [email protected].
5
    State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Science, School of Public Health, Xiamen University, 361102, Xiamen, P. R. China. [email protected].

Abstract

The human liver chimeric mouse with primary human hepatocytes (PHHs) engraftment has been demonstrated to be a useful animal model to study hepatitis B virus (HBV) pathogenesis and evaluate anti-HBV drugs. However, the disadvantages of using PHHs include the inability for cellular expansion in vitro, limited donor availability, individual differences, and ethical issues, necessitating the development of alternatives. To obtain in vitro expandable hepatocytes, we optimized the hepatic differentiation procedure of the human liver progenitor cell line, HepaRG, using four functional small molecules (4SM) and enriched the precursor hepatocyte-like cells (HLCs). HepaRG cells of different hepatic differentiation states were engrafted to immunodeficient mice (FRGS) with weekly 4SM treatment. The HepaRG-engrafted mice were challenged with HBV and/or treated with several antivirals to evaluate their effects. We demonstrated that the 4SM treatment enhanced hepatic differentiation and promoted cell proliferation capacity both in vitro and in vivo. Mice engrafted with enriched HepaRG of prehepatic differentiation and treated with 4SM displayed approximately 10% liver chimerism at week 8 after engraftment and were maintained at this level for another 16 weeks. Therefore, we developed a HepaRG-based human liver chimeric mouse model: HepaRG-FRGS. Our experimental results showed that the liver chimerism of the mice was adequate to support chronic HBV infection for 24 weeks and to evaluate antivirals. We also demonstrated that HBV infection in HepaRG cells was dependent on their hepatic differentiation state and liver chimerism in vivo. Overall, HepaRG-FRGS mice provide a novel human liver chimeric mouse model to study chronic HBV infection and evaluate anti-HBV drugs.

PMID:
    30097574
DOI:
    10.1038/s41426-018-0143-9

StephenW

Emerg Microbes Infect。 2018年8月10日; 7（1）：144。 doi：10.1038 / s41426-018-0143-9。
优化的HepaRG是产生用于慢性乙型肝炎病毒感染的人肝脏嵌合小鼠模型的合适细胞来源。
Yuan L1，Liu X1，Zhang L1，Zhang Y1，Chen Y1，Li X1，Wu K1，Cao J1，Hou W1，Que Y1，Zhang J1，Zhu H2，Yuan Q3，Tang Q4，Cheng T5，Xia N1。
作者信息

1
    厦门大学公共卫生学院生命科学学院传染病诊断与疫苗研究所分子疫苗学与分子诊断国家重点实验室，361102，厦门，中国。
2
    罗格斯大学新泽西医学院微生物学，生物化学和分子遗传学系，225 Warren Street，Newark，NJ，070101，USA。
3
    厦门大学公共卫生学院生命科学学院传染病诊断与疫苗研究所分子疫苗学与分子诊断国家重点实验室，361102，厦门，中国。 [email protected]。
4
    华盛顿特区，霍华德大学医学院微生物学系，20059，美国。 [email protected]。
五
    厦门大学公共卫生学院生命科学学院传染病诊断与疫苗研究所分子疫苗学与分子诊断国家重点实验室，361102，厦门，中国。 [email protected]。

抽象

具有原代人肝细胞（PHH）植入的人肝嵌合小鼠已被证明是研究乙型肝炎病毒（HBV）发病机理和评估抗HBV药物的有用动物模型。然而，使用PHH的缺点包括体外细胞扩增不能，供体可用性有限，个体差异和伦理问题，需要开发替代品。为了获得体外可扩增的肝细胞，我们使用四种功能性小分子（4SM）优化了人肝脏祖细胞系HepaRG的肝分化程序，并富集了前体肝细胞样细胞（HLC）。将具有不同肝分化状态的HepaRG细胞移植到免疫缺陷小鼠（FRGS）中，每周4SM处理。用HBV攻击HepaRG-移植的小鼠和/或用几种抗病毒剂处理以评估它们的作用。我们证明了4SM处理增强了肝脏分化并促进了体外和体内的细胞增殖能力。植入富含肝前分化的HepaRG并用4SM处理的小鼠在植入后第8周显示约10％的肝嵌合体，并在该水平维持另外16周。因此，我们开发了基于HepaRG的人肝嵌合小鼠模型：HepaRG-FRGS。我们的实验结果表明，小鼠的肝嵌合体足以支​​持慢性HBV感染24周并评估抗病毒药物。我们还证明了HepaRG细胞中的HBV感染依赖于它们的肝分化状态和体内肝脏嵌合状态。总体而言，HepaRG-FRGS小鼠提供了一种新的人肝脏嵌合小鼠模型，用于研究慢性HBV感染并评估抗HBV药物。

结论：
    30097574
DOI：
    10.1038 / s41426-018-0143-9