Single hepatocytes show persistence and transcriptional inactivity of hepatitis B
Single hepatocytes show persistence and transcriptional inactivity of hepatitis B
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DOI：
10.1172/jci.insight.140584
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发表时间：
2020-10-02
期刊：
影响因子：
8
通讯作者：
Thio, Chloe L.
中科院分区：
文献类型：
--
作者：
Balagopal, Ashwin;Grudda, Tanner;Thio, Chloe L.
超过2.7亿慢性HBV感染者无法治愈。核苷（酸）类似物（NUC）是抗HBV治疗的主要药物，可阻断HBV逆转录。NUC不消除核内共价闭合环状DNA（cccDNA），病毒RNA（包括前基因组RNA（pgRNA））从所述核内共价闭合环状DNA转录。设计治疗方法的一个关键差距是了解NUC如何影响HBV复制和转录，因为血清标志物对肝内HBV的认识不完整。我们应用单细胞激光捕获显微切割和液滴数字PCR技术对5例服用NUC 2-4年以上的HBV/HIV合并感染者进行配对肝活检。从活检1到2，HBV感染肝细胞的比例随着NUC治疗的依从性而下降（P < 0.05）;我们推断，在这些参与者中，NUC根除HBV将需要10年以上。在单个肝细胞中，pgRNA水平在NUC治疗期间降低28- 73倍，对应于组织HBV核心抗原染色降低（P < 0.01）。在5名参与者中的4名中，存在具有cccDNA但不可检测的pgRNA（转录失活）的肝细胞，并且这些在NUC治疗期间在3名参与者中富集。解开cccDNA转录失活机制的进一步工作可能导致可以在所有肝细胞中实现这一点的疗法，从而导致功能性治愈。
There is no cure for the more than 270 million people chronically infected with HBV. Nucleos(t) ide analogs (NUCs), the mainstay of anti-HBV treatment, block HBV reverse transcription. NUCs do not eliminate the intranuclear covalently closed circular DNA (cccDNA), from which viral RNAs, including pregenomic RNA (pgRNA), are transcribed. A key gap in designing a cure is understanding how NUCs affect HBV replication and transcription because serum markers yield an incomplete view of intrahepatic HBV. We applied single-cell laser capture microdissection and droplet digital PCR to paired liver biopsies collected from 5 HBV/HIV-coinfected persons who took NUCs over 2-4 years. From biopsy 1 to 2, proportions of HBV-infected hepatocytes declined with adherence to NUC treatment (P < 0.05); we extrapolated that eradication of HBV will take over 10 decades with NUCs in these participants. In individual hepatocytes, pgRNA levels diminished 28- to 73-fold during NUC treatment, corresponding with decreased tissue HBV core antigen staining (P < 0.01). In 4 out of 5 participants, hepatocytes with cccDNA but undetectable pgRNA (transcriptionally inactive) were present, and these were enriched in 3 participants during NUC treatment. Further work to unravel mechanisms of cccDNA transcriptional inactivation may lead to therapies that can achieve this in all hepatocytes, resulting in a functional cure.