Integrated hepatitis B virus DNA maintains surface antigen production during antiviral treatment
Tanner Grudda, … , Ashwin Balagopal, Chloe L. Thio
Tanner Grudda, … , Ashwin Balagopal, Chloe L. Thio
Published July 7, 2022
Citation Information: J Clin Invest. 2022;132(18):e161818. https://doi.org/10.1172/JCI161818.
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Research Article Infectious disease Virology

Integrated hepatitis B virus DNA maintains surface antigen production during antiviral treatment

Abstract

The focus of hepatitis B functional cure, defined as sustained loss of hepatitis B virus (HBV) surface antigen (HBsAg) and HBV DNA from blood, is on eliminating or silencing the intranuclear template for HBV replication, covalently closed circular DNA (cccDNA). However, HBsAg also derives from HBV DNA integrated into the host genome (iDNA). Little is known about the contribution of iDNA to circulating HBsAg with current therapeutics. We applied a multiplex droplet digital PCR assay to demonstrate that iDNA is responsible for maintaining HBsAg quantities in some individuals. Using paired bulk liver tissue from 16 HIV/HBV-coinfected persons on nucleos(t)ide analog (NUC) therapy, we demonstrate that people with larger HBsAg declines between biopsies derive HBsAg from cccDNA, whereas people with stable HBsAg levels derive predominantly from iDNA. We applied our assay to individual hepatocytes in paired tissues from 3 people and demonstrated that the individual with significant HBsAg decline had a commensurate loss of infected cells with transcriptionally active cccDNA, while individuals without HBsAg decline had stable or increasing numbers of cells producing HBsAg from iDNA. We demonstrate that while NUC therapy may be effective at controlling cccDNA replication and transcription, innovative treatments are required to address iDNA transcription that sustains HBsAg production.

Authors

Tanner Grudda, Hyon S. Hwang, Maraake Taddese, Jeffrey Quinn, Mark S. Sulkowski, Richard K. Sterling, Ashwin Balagopal, Chloe L. Thio

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Figure 6

HBV serology and single-cell viral transcriptional landscapes from 3 representative people over time.

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HBV serology and single-cell viral transcriptional landscapes from 3 rep...
(A) Serum HBV DNA (dark green) and serum HBsAg (slate) levels between biopsies are represented for each person who underwent single-cell examination (n = 3). All available time points are shown and open circles are those that were below the limit of detection for each assay. Vertical dashed lines indicate when biopsies were taken from each person. (B) Each data point represents a single hepatocyte after QC exclusion. Grids are arrayed in the x-y plane to reflect their position in a contiguous section of tissue. Three representative people are shown at each biopsy. The size of each data point corresponds to the quantity of HBV transcripts/cell, as measured by the mid-HBV assay, and open points indicate that no HBV transcription was detected. Colors indicate the source of viral transcripts in cells, as specified in the legend and determined by the iDNA TI. Some cells were found to have cccDNA-derived transcription but had undetectable S transcripts; these were colored gold and are shown as the smallest points. All cells that were deemed analyzable (because they passed QC by having sufficient total RNA) are depicted, irrespective of their infection or viral transcriptional status.