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Differential Intrahepatic Integrated HBV DNA Patterns Between HBeAg-Positive and HBeAg-Negative Chronic Hepatitis B

Daryl T-Y Lau, Elena S Kim, Zhili Wang

    Medrxiv : the Preprint Server for Health Sciences
    |March 17, 2025
    PubMed

    Insights

    Integrated HBV DNA (iDNA) is a significant source of HBsAg in HBeAg(-) chronic hepatitis B (CHB) patients. This study highlights iDNA

    Area of Science:

    • Hepatology
    • Virology
    • Molecular Biology

    Background:

    • Hepatitis B surface antigen (HBsAg) can originate from intrahepatic covalently closed circular DNA (cccDNA) and integrated HBV DNA (iDNA).
    • Understanding the relative contributions of cccDNA and iDNA to HBsAg production is crucial for chronic hepatitis B (CHB) management.

    Purpose of the Study:

    • To investigate the role of intrahepatic HBV DNA forms, specifically cccDNA and iDNA, in HBsAg production in treatment-naive CHB patients.
    • To compare the characteristics of iDNA and cccDNA in HBeAg-positive (HBeAg+) and HBeAg-negative (HBeAg-) CHB participants.

    Main Methods:

    • Analysis of liver tissues from 24 HBeAg(+) and 32 HBeAg(-) treatment-naive CHB participants.
    • Quantification of intrahepatic cccDNA using qPCR after selective digestion.
    • Detection and characterization of intrahepatic iDNA using HBV hybridization-targeted next-generation sequencing (HBV-NGS) and ChimericSeq for junction analysis.

    Main Results:

    • HBeAg(+) participants showed higher levels of intrahepatic cccDNA, serum HBV DNA, HBV RNA, HBcrAg, and qHBsAg compared to HBeAg(-) participants.
    • HBsAg staining was prevalent in over 85% of livers in both HBeAg(+) and HBeAg(-) groups.
    • HBeAg(-) participants predominantly had iDNA (>50% of total HBV DNA), while HBeAg(+) participants had a more balanced or cccDNA-dominant profile; iDNA integration patterns differed between groups, with clustering at DR2-DR1 in HBeAg(-) livers.

    Conclusions:

    • Serum HBV RNA and HBcrAg levels correlate with intrahepatic cccDNA concentrations.
    • In HBeAg(-) CHB patients, high levels of intrahepatic iDNA and HBsAg, despite lower cccDNA, suggest iDNA is the primary source of HBsAg.
    • The distinct integration patterns of iDNA may reflect different mechanisms of HBV DNA integration in HBeAg(+) versus HBeAg(-) CHB.
    Abstract

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