Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNA

Full-length hepatitis B virus (HBV) transcripts of chimpanzees and patients treated with multidose (MD) HBV siRNA ARC-520 and entecavir (ETV) were characterized by single-molecule real-time (SMRT) sequencing, identifying multiple types of transcripts with the potential to encode HBx, HBsAg, HBeAg, c...

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Main Authors: Christine I. Wooddell, Dean Sanders, Zhao Xu, Lung-Yi Mak, Thomas Schluep, Wai-Kay Seto, Bruce D. Given, Man-Fung Yuen
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Viruses
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Online Access:https://www.mdpi.com/1999-4915/16/12/1943
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author Christine I. Wooddell
Dean Sanders
Zhao Xu
Lung-Yi Mak
Thomas Schluep
Wai-Kay Seto
Bruce D. Given
Man-Fung Yuen
author_facet Christine I. Wooddell
Dean Sanders
Zhao Xu
Lung-Yi Mak
Thomas Schluep
Wai-Kay Seto
Bruce D. Given
Man-Fung Yuen
author_sort Christine I. Wooddell
collection DOAJ
description Full-length hepatitis B virus (HBV) transcripts of chimpanzees and patients treated with multidose (MD) HBV siRNA ARC-520 and entecavir (ETV) were characterized by single-molecule real-time (SMRT) sequencing, identifying multiple types of transcripts with the potential to encode HBx, HBsAg, HBeAg, core, and polymerase, as well as transcripts likely to be derived from dimers of dslDNA, and these differed between HBeAg-positive (HBeAg+) and HBeAg-negative (HBeAg−) individuals. HBV transcripts from the last follow-up ~30 months post-ARC-520 treatment were categorized from one HBeAg+ (one of two previously highly viremic patients that became HBeAg− upon treatment and had greatly reduced cccDNA products) and four HBeAg− patients. The previously HBeAg+ patient received a biopsy that revealed that he had 3.4 copies/cell cccDNA (two to three orders of magnitude more cccDNA than HBeAg− chimpanzees) but expressed primarily truncated X and HBsAg from iDNA, like two patients that were HBeAg− at the start of the study and had one copy/cell cccDNA. No HBV transcripts were detected in two other HBeAg− patients that had ~0.3 copies/cell cccDNA, one of which had seroconverted for HBsAg. The paucity of cccDNA-derived transcripts in the presence of high cccDNA demonstrates the transcriptional silencing of HBV following MD siRNA treatment with ETV.
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spelling doaj-art-f4ec0c584a2e4a2e927187c3b8e323a02024-12-27T14:59:19ZengMDPI AGViruses1999-49152024-12-011612194310.3390/v16121943Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNAChristine I. Wooddell0Dean Sanders1Zhao Xu2Lung-Yi Mak3Thomas Schluep4Wai-Kay Seto5Bruce D. Given6Man-Fung Yuen7Arrowhead Pharmaceuticals Inc., 502 S. Rosa Road, Madison, WI 53719, USAArrowhead Pharmaceuticals Inc., 502 S. Rosa Road, Madison, WI 53719, USAArrowhead Pharmaceuticals Inc., 10102 Hoyt Park Drive, San Diego, CA 92131, USADepartment of Medicine & State Key Laboratory of Liver Research, School of Clinical Medicine, The University of Hong Kong, Hong Kong, ChinaArrowhead Pharmaceuticals Inc., 177 E. Colorado Boulevard, Suite 700, Pasadena, CA 91105, USADepartment of Medicine & State Key Laboratory of Liver Research, School of Clinical Medicine, The University of Hong Kong, Hong Kong, ChinaArrowhead Pharmaceuticals Inc., 177 E. Colorado Boulevard, Suite 700, Pasadena, CA 91105, USADepartment of Medicine & State Key Laboratory of Liver Research, School of Clinical Medicine, The University of Hong Kong, Hong Kong, ChinaFull-length hepatitis B virus (HBV) transcripts of chimpanzees and patients treated with multidose (MD) HBV siRNA ARC-520 and entecavir (ETV) were characterized by single-molecule real-time (SMRT) sequencing, identifying multiple types of transcripts with the potential to encode HBx, HBsAg, HBeAg, core, and polymerase, as well as transcripts likely to be derived from dimers of dslDNA, and these differed between HBeAg-positive (HBeAg+) and HBeAg-negative (HBeAg−) individuals. HBV transcripts from the last follow-up ~30 months post-ARC-520 treatment were categorized from one HBeAg+ (one of two previously highly viremic patients that became HBeAg− upon treatment and had greatly reduced cccDNA products) and four HBeAg− patients. The previously HBeAg+ patient received a biopsy that revealed that he had 3.4 copies/cell cccDNA (two to three orders of magnitude more cccDNA than HBeAg− chimpanzees) but expressed primarily truncated X and HBsAg from iDNA, like two patients that were HBeAg− at the start of the study and had one copy/cell cccDNA. No HBV transcripts were detected in two other HBeAg− patients that had ~0.3 copies/cell cccDNA, one of which had seroconverted for HBsAg. The paucity of cccDNA-derived transcripts in the presence of high cccDNA demonstrates the transcriptional silencing of HBV following MD siRNA treatment with ETV.https://www.mdpi.com/1999-4915/16/12/1943hepatitis B virus transcriptionhepatitis B virus RNAHBV XcccDNAchronic hepatitis BHBV siRNA
spellingShingle Christine I. Wooddell
Dean Sanders
Zhao Xu
Lung-Yi Mak
Thomas Schluep
Wai-Kay Seto
Bruce D. Given
Man-Fung Yuen
Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNA
Viruses
hepatitis B virus transcription
hepatitis B virus RNA
HBV X
cccDNA
chronic hepatitis B
HBV siRNA
title Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNA
title_full Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNA
title_fullStr Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNA
title_full_unstemmed Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNA
title_short Characterization of Hepatitis B Virus Transcripts in Chronically HBV-Infected Chimpanzees and Patients Treated with ARC-520 siRNA Demonstrates Transcriptional Silencing of cccDNA
title_sort characterization of hepatitis b virus transcripts in chronically hbv infected chimpanzees and patients treated with arc 520 sirna demonstrates transcriptional silencing of cccdna
topic hepatitis B virus transcription
hepatitis B virus RNA
HBV X
cccDNA
chronic hepatitis B
HBV siRNA
url https://www.mdpi.com/1999-4915/16/12/1943
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