单肝细胞显示出乙型肝炎的持久性和转录失活

StephenW

Single hepatocytes show persistence and transcriptional inactivity of hepatitis B
Ashwin Balagopal  1 , Tanner Grudda  1 , Ruy M Ribeiro  2   3 , Yasmeen S Saad  1 , Hyon S Hwang  1 , Jeffrey Quinn  1 , Michael Murphy  1 , Kathleen Ward  1 , Richard K Sterling  4 , Yang Zhang  5 , Alan S Perelson  2 , Mark S Sulkowski  1 , William O Osburn  1 , Chloe L Thio  1
Affiliations
Affiliations

    1
    Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
    2
    Theoretical Biology and Biophysics Group, Los Alamos National Laboratory, Los Alamos, New Mexico, USA.
    3
    Faculdade de Medicina Universidade de Lisboa, Lisbon, Portugal.
    4
    Division of Gastroenterology, Hepatology and Nutrition, Virginia Commonwealth University, Richmond, Virginia, USA.
    5
    Department of Pathology, Johns Hopkins Hospital, Baltimore, Maryland, USA.

    PMID: 33004689 DOI: 10.1172/jci.insight.140584

Abstract

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.

Keywords: Hepatitis; Hepatology; Infectious disease; Molecular biology; Transcription.

StephenW

单肝细胞显示出乙型肝炎的持久性和转录失活
Ashwin Balagopal 1，Tanner Grudda 1，Ruy M Ribeiro 2 3，Yasmeen S Saad 1，Hyon S Hwang 1，Jeffrey Quinn 1，Michael Murphy 1，Kathleen Ward 1，Richard K Sterling 4，Yang Zhang 5，Alan S Perelson 2， Mark S Sulkowski 1，William O Osburn 1，Chloe L Thio 1
隶属关系
隶属关系

    1个
    美国马里兰州巴尔的摩市约翰霍普金斯大学医学院医学系。
    2
    美国新墨西哥州洛斯阿拉莫斯国家生物学理论生物学和生物物理学组。
    3
    Faculdade de Medicina大学里斯本，葡萄牙里斯本。
    4
    美国弗吉尼亚州里士满，弗吉尼亚联邦大学胃肠病学，肝病学和营养学系。
    5
    美国马里兰州巴尔的摩市约翰霍普金斯医院病理科。

    PMID：33004689 DOI：10.1172 / jci.insight.140584

抽象

超过2.7亿慢性感染HBV的人无法治愈。抗-HBV治疗的主要手段-核苷类似物（NUCs）阻止HBV逆转录。 NUC没有消除核内共价封闭的环状DNA（cccDNA），从中转录出病毒RNA，包括前基因组RNA（pgRNA）。设计治疗方法的关键差距在于了解NUC如何影响HBV复制和转录，因为血清标志物不能完全反映肝内HBV。我们将单细胞激光捕获显微切割技术和液滴数字PCR技术应用于从5名HBV / HIV合并感染者中收集的成对肝活检样本，这些人在2-4年内服用了NUC。从活检1至2，坚持NUC治疗后，感染HBV的肝细胞比例下降（P <0.05）。我们推断，在这些参与者中，使用NUC消灭HBV将需要10多年。在单个肝细胞中，在NUC治疗过程中pgRNA水平降低了28到73倍，与组织HBV核心抗原染色减少相对应（P <0.01）。在五分之四的参与者中，存在具有cccDNA但未检测到pgRNA（转录失活）的肝细胞，并且在NUC治疗过程中有三名参与者富集了这些肝细胞。进一步阐明cccDNA转录失活机制的工作可能导致可以在所有肝细胞中实现这一目标的疗法，从而实现功能性治愈。

关键字：肝炎；肝病学传染病;分子生物学;转录。

StephenW

https://insight.jci.org/articles/view/140584