Rapid Turnover of Hepatitis B Virus Covalently Closed Circular DNA Indicated by Monitoring Emergence and Reversion of Signature-Mutation in Treated Chronic Hepatitis B Patients

Qi Huang, Bin Zhou, Dawei Cai, Yuhua Zong, Yaobo Wu, Shi Liu, Alexandre Mercier, Haitao Guo, Jinlin Hou, Richard Colonno, Jian Sun, Qi Huang, Bin Zhou, Dawei Cai, Yuhua Zong, Yaobo Wu, Shi Liu, Alexandre Mercier, Haitao Guo, Jinlin Hou, Richard Colonno, Jian Sun

Abstract

Background and aims: 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 strategies to clear HBV by fully blocking the de novo generation of cccDNA.

Approach and results: 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 virological 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.

Conclusions: 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.

Trial registration: ClinicalTrials.gov NCT00962533.

© 2020 The Authors. Hepatology published by Wiley Periodicals LLC on behalf of American Association for the Study of Liver Diseases.

Figures

Fig. 1
Fig. 1
Flowchart of patient selection. Abbreviation: ADV, adefovir dipivoxil.
Fig. 2
Fig. 2
Genetic composition of HBV cccDNA, DNA, and RNA. Percentage of LAMR rtM204I/V mutation in serum and liver biopsy samples of 5 EFFORT patients collected at week 104 and 2 ML18376 patients collected at baseline (A). The correlation of LAMR composition among intrahepatic RNA, intrahepatic DNA, cccDNA, and serum HBV RNA were analyzed (B‐E).
Fig. 3
Fig. 3
Dynamic change of LAMR in serum HBV RNA and DNA during the development of VB in the EFFORT study (patients 7, 17, 20, 26, and 31). Abbreviations: ADV, adefovir dipivoxil; HBsAg, hepatitis B surface antigen; Ig, immunoglobulin; LdT, telbivudine.
Fig. 4
Fig. 4
Dynamic change of LAMR in serum HBV RNA and DNA during the reversion of LAMR in ML18376 study (patients 13, 45, 50, 53, and 194). Abbreviation: Ig, immunoglobulin.
Fig. 5
Fig. 5
cccDNA turnover model. In the proposed model, the genetic composition of serum HBV RNA, a genetic biomarker for cccDNA turnover, changes within several months, which is much faster than previously thought. Abbreviation: pgRNA, pregenomic RNA.

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Source: PubMed

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