通过监测已治疗的慢性乙型肝炎患者的特征突变的出现和逆

StephenW

Hepatology. 2020 Mar 19. doi: 10.1002/hep.31240. [Epub ahead of print]
Rapid Turnover of HBV cccDNA Indicated by Monitoring Emergence and Reversion of Signature-Mutation in Treated Chronic Hepatitis B Patients.
Huang Q1, Zhou B2,3, Cai D1, Zong Y1, Wu Y2, Liu S2, Mercier A1, Guo H3,4, Hou J2, Colonno R1, Sun J2.
Author information

1
    Assembly Biosciences, Inc, South San Francisco, CA, United States.
2
    State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Viral Hepatitis Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China.
3
    Department of Microbiology and Immunology, Indiana University, Indianapolis, IN, United States.
4
    Cancer Virology Program, UPMC Hillman Cancer Center, Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA, United States.

Abstract

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) plays a pivotal role in the establishment and persistence of HBV infection. Understanding the turnover time of preexisting cccDNA pools would be helpful in designing new strategies to clear HBV by fully blocking the de novo generation of cccDNA. In this study, we retrospectively monitored the emergence and reversion of the rtM204I/V mutant, a signature lamivudine resistance (LAMR ) mutation serving as a biomarker of cccDNA turnover in liver biopsies and longitudinal serum samples from two clinical trials. Methodologies were optimized to differentially isolate and sequence HBV virion DNA, cccDNA, and HBV RNA from clinical samples. A strong correlation was observed between LAMR composition of cccDNA with that of serum and intrahepatic HBV RNA in paired liver and serum samples (r=0.96 and 0.90 respectively), suggesting that serum HBV RNA can serve as a surrogate marker of cccDNA genetic composition when liver biopsies are unavailable. LAMR mutations emerged and increased from undetectable to 40%~90% within 16-28 weeks in serum HBV RNA from telbivudine-treated patients experiencing virologic breakthrough. Similarly, in lamivudine-resistant patients who switched to interferon therapy, serum HBV RNA population bearing 100% LAMR mutations fully reversed back to wild type within 24~48 weeks. Conclusion: The genetic composition dynamics of serum HBV RNA and biopsy cccDNA in treated HBV patients indicates that cccDNA turnover occurs relatively rapidly (several months), offering a possibility of HBV cure with finite therapy through completely blocking cccDNA replenishment.

This article is protected by copyright. All rights reserved.
KEYWORDS:

Genetic marker; HBV RNA; Nuc resistance; cccDNA half-life

PMID:
    32189364
DOI:
    10.1002/hep.31240

StephenW

肝病学。 2020年3月19日。doi：10.1002 / hep.31240。 [Epub提前发行]
通过监测已治疗的慢性乙型肝炎患者的特征突变的出现和逆转，可以指示HBV cccDNA的快速周转。
黄Q1，周B2,3，蔡D1，宗Y1，吴Y2，刘S2，Mercier A1，郭H3,4，侯J2，科洛诺R1，孙J2。
作者信息

1个
    美国加利福尼亚州南旧金山，Assembly Biosciences，Inc。
2
    南方医科大学南方医院传染病科，器官衰竭研究国家重点实验室，广东省病毒性肝炎研究重点实验室，广州。
3
    美国印第安纳州印第安纳波利斯的印第安纳大学微生物学和免疫学系。
4
    美国宾夕法尼亚州匹兹堡大学匹兹堡大学微生物学和分子遗传学系，UPMC Hillman癌症中心癌症病毒学计划。

抽象

乙型肝炎病毒（HBV）共价闭合环状DNA（cccDNA）在HBV感染的建立和持续中起着关键作用。了解预先存在的cccDNA池的周转时间将有助于设计通过完全阻断cccDNA从头产生来清除HBV的新策略。在这项研究中，我们回顾性地监测了rtM204I / V突变体的出现和回复，该突变体是标志性拉米夫定抗性（LAMR）突变，是肝活检和纵向血清样品中cccDNA更新的生物标志物，来自两个临床试验。优化方法以从临床样品中差异分离和测序HBV病毒粒子DNA，cccDNA和HBV RNA。在成对的肝脏和血清样品中，cccDNA的LAMR组成与血清和肝内HBV RNA的含量之间存在很强的相关性（分别为r = 0.96和0.90），这表明肝脏中的血清HBV RNA可以作为cccDNA遗传组成的替代标志物无法进行活检。在接受病毒学突破的替比夫定治疗的患者的血清HBV RNA中，在16-28周内，LAMR突变出现并从不可检测的增加到40％〜90％。同样，在接受干扰素治疗的拉米夫定耐药患者中，带有100％LAMR突变的血清HBV RNA群体在24〜48周内完全回复为野生型。结论：治疗的HBV患者血清HBV RNA和活检cccDNA的遗传组成动态表明，cccDNA周转发生相对较快（数月），这可能通过完全阻断cccDNA的补充而通过有限的疗法治愈HBV。

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关键字：

遗传标记HBV RNA；抗核cccDNA半衰期

PMID：
    32189364
DOI：
    10.1002 / hep.31240