Human induced-pluripotent stem cell-derived hepatocyte-like cells as an in vitro model of human hepatitis B virus infection.
Human induced-pluripotent stem cell-derived hepatocyte-like cells as an in vitro model of human hepatitis B virus infection.
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DOI：
10.1038/srep45698
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发表时间：
2017-04-04
影响因子：
4.6
通讯作者：
Mizuguchi H
中科院分区：
文献类型：
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作者：
Sakurai F;Mitani S;Yamamoto T;Takayama K;Tachibana M;Watashi K;Wakita T;Iijima S;Tanaka Y;Mizuguchi H
为了了解B型肝炎病毒（HBV）的生命周期，并开发有效的抗HBV药物，一个有用的体外细胞培养系统，使HBV感染和再现病毒-宿主相互作用是必不可少的;然而，预先存在的体外HBV感染模型往往是有问题的。在这里，我们研究了人诱导多能干（iPS）细胞衍生的肝细胞样细胞（iPS-HLC）作为体外HBV感染模型的潜力。参与HBV感染的几个基因的表达水平，包括牛磺胆酸钠共转运多肽（NTCP）基因，随着人iPS细胞向iPS-HLCs的分化状态逐渐升高。这些基因的mRNA水平在原代人肝细胞（PHH）和iPS-HLC之间相当。在用HBV接种后，我们发现在iPS-HLC中显著产生HBV蛋白和病毒RNA。通过Southern印迹分析在iPS-HLC中检测到HBV基因组的三种主要形式。抗HBV药物恩替卡韦和Myrcludex-B，这是一种核苷类似物逆转录酶抑制剂和合成的前S1肽，分别显着抑制iPS-HLCs中的HBV感染。这些数据表明iPS-HLC可以用作有希望的体外HBV感染模型。
In order to understand the life cycle of hepatitis B virus (HBV) and to develop efficient anti-HBV drugs, a useful in vitro cell culture system which allows HBV infection and recapitulates virus-host interactions is essential; however, pre-existing in vitro HBV infection models are often problematic. Here, we examined the potential of human induced-pluripotent stem (iPS) cell-derived hepatocyte-like cells (iPS-HLCs) as an in vitro HBV infection model. Expression levels of several genes involved in HBV infection, including the sodium taurocholate cotransporting polypeptide (NTCP) gene, were gradually elevated as the differentiation status of human iPS cells proceeded to iPS-HLCs. The mRNA levels of these genes were comparable between primary human hepatocytes (PHHs) and iPS-HLCs. Following inoculation with HBV, we found significant production of HBV proteins and viral RNAs in iPS-HLCs. The three major forms of the HBV genome were detected in iPS-HLCs by Southern blotting analysis. Anti-HBV agents entecavir and Myrcludex-B, which are a nucleoside analogue reverse transcriptase inhibitor and a synthetic pre-S1 peptide, respectively, significantly inhibited HBV infection in iPS-HLCs. These data demonstrate that iPS-HLCs can be used as a promising in vitro HBV infection model.