Hepatitis B virus (HBV) can manipulate the microRNA (miRNA) regulatory networks in infected cells to create a permissive environment for viral replication, cellular injury, disease onset, and its progression. The aim of the present study was to understand the miRNA networks and their target genes in the liver of hepatitis B patients involved in HBV replication, liver injury, and liver fibrosis. We investigated differentially expressed miRNAs by microarray in liver biopsy samples from different stages of HBV infection and liver disease (immune‐tolerant [n = 8], acute viral hepatitis [n = 8], no fibrosis [n = 16], early [F1+F2, n = 19] or late [F3+F4, n = 14] fibrosis, and healthy controls [n = 7]). miRNA expression levels were analyzed by unsupervised principal component analysis and hierarchical clustering. Analysis of miRNA–mRNA regulatory networks identified 17 miRNAs and 18 target gene interactions with four distinct nodes, each representing a stage‐specific gene regulation during disease progression. The immune‐tolerant group showed elevated miR‐199a‐5p, miR‐221‐3p, and Let‐7a‐3p levels, which could target genes involved in innate immune response and viral replication. In the acute viral hepatitis group, miR‐125b‐5p and miR‐3613‐3p were up, whereas miR‐940 was down, which might affect cell proliferation through the signal transducer and activator of transcription 3 pathway. In early fibrosis, miR‐34b‐3p, miR‐1224‐3p, and miR‐1227‐3p were up, while miR‐499a‐5p was down, which together possibly mediate chronic inflammation. In advanced fibrosis, miR‐1, miR‐10b‐5p, miR‐96‐5p, miR‐133b, and miR‐671‐5p were up, while miR‐20b‐5p and miR‐455‐3p were down, possibly allowing chronic disease progression. Interestingly, only 8 of 17 liver‐specific miRNAs exhibited a similar expression pattern in patient sera. Conclusion: miRNA signatures identified in this study corroborate previous findings and provide fresh insight into the understanding of HBV‐associated liver diseases which may be helpful in developing early‐stage disease diagnostics and targeted therapeutics. (Hepatology 2018;67:1695‐1709)

中文翻译：

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乙型肝炎病毒感染患者肝活检中的整体微 RNA 表达谱表明病毒持续存在和肝细胞损伤的特定 miRNA 特征

乙型肝炎病毒 (HBV) 可以操纵受感染细胞中的 microRNA (miRNA) 调节网络，为病毒复制、细胞损伤、疾病发作及其进展创造一个允许的环境。本研究的目的是了解参与 HBV 复制、肝损伤和肝纤维化的乙型肝炎患者肝脏中的 miRNA 网络及其靶基因。我们通过微阵列研究了来自 HBV 感染和肝脏疾病不同阶段（免疫耐受 [n = 8]、急性病毒性肝炎 [n = 8]、无纤维化 [n = 16]、早期 [F1 +F2，n = 19] 或晚期 [F3+F4，n = 14] 纤维化和健康对照 [n = 7]）。通过无监督主成分分析和层次聚类分析 miRNA 表达水平。对 miRNA-mRNA 调控网络的分析确定了 17 个 miRNA 和 18 个靶基因与四个不同节点的相互作用，每个节点代表疾病进展过程中的特定阶段基因调控。免疫耐受组显示 miR-199a-5p、miR-221-3p 和 Let-7a-3p 水平升高，这可能靶向参与先天免疫反应和病毒复制的基因。在急性病毒性肝炎组，miR-125b-5p和miR-3613-3p上升，而miR-940下降，可能通过信号转导和转录激活因子3通路影响细胞增殖。在纤维化早期，miR-34b-3p、miR-1224-3p和miR-1227-3p上升，而miR-499a-5p下降，这可能共同介导慢性炎症。在晚期纤维化中，miR-1、miR-10b-5p、miR-96-5p、miR-133b和miR-671-5p上升，而 miR-20b-5p 和 miR-455-3p 下降，可能允许慢性疾病进展。有趣的是，17 种肝脏特异性 miRNA 中只有 8 种在患者血清中表现出相似的表达模式。结论：本研究中鉴定的 miRNA 特征证实了先前的发现，并为了解 HBV 相关肝病提供了新的见解，这可能有助于开发早期疾病诊断和靶向治疗。（肝病学 2018 年；67：1695-1709）本研究中鉴定的 miRNA 特征证实了先前的发现，并为了解 HBV 相关肝病提供了新的见解，这可能有助于开发早期疾病诊断和靶向治疗。（肝病学 2018 年；67：1695-1709）本研究中鉴定的 miRNA 特征证实了先前的发现，并为了解 HBV 相关肝病提供了新的见解，这可能有助于开发早期疾病诊断和靶向治疗。（肝病学 2018 年；67：1695-1709）