{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T20:22:01Z","timestamp":1791836521330,"version":"4.3.4"},"reference-count":52,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2021,3,30]],"date-time":"2021-03-30T00:00:00Z","timestamp":1617062400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Chronic hepatitis B infection remains a globally important cause of morbidity and mortality and has recently undergone a renaissance in therapeutic interest with increased pre-clinical and clinical testing of new drug classes. One of the first new classes in the clinic was RNA interference agents, which have the potential to impact the entire viral life cycle by reducing all virus-produced mRNA. Early clinical testing with the first of these agents in the clinic, ARC-520, demonstrated that rapid and deep reductions in viral proteins, RNA and DNA could be produced with this approach, but also the surprising insight that HBsAg production from incomplete HBV DNA integrated into the host genome appears to play a heretofore unappreciated and important role in maintaining circulating HBsAg, thought to play a fundamental role in preventing host clearance of the virus. Thus, accounting for viral DNA integration in novel HBV treatment approaches may prove to be essential to achieving successful finite therapies of this difficult to treat chronic infection.<\/jats:p>","DOI":"10.3390\/v13040581","type":"journal-article","created":{"date-parts":[[2021,3,31]],"date-time":"2021-03-31T00:13:10Z","timestamp":1617149590000},"page":"581","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["RNA Interference Therapy for Chronic Hepatitis B Predicts the Importance of Addressing Viral Integration When Developing Novel Cure Strategies"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8203-7493","authenticated-orcid":false,"given":"Christine I.","family":"Wooddell","sequence":"first","affiliation":[{"name":"Arrowhead Pharmaceuticals, 502 South Rosa Road, Madison, WI 53719, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1150-5840","authenticated-orcid":false,"given":"Adam J.","family":"Gehring","sequence":"additional","affiliation":[{"name":"Toronto Centre for Liver Disease, Toronto General Hospital Research Institute, University Health Network, Toronto, ON M5G 1L7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7985-7725","authenticated-orcid":false,"given":"Man-Fung","family":"Yuen","sequence":"additional","affiliation":[{"name":"Department of Medicine, The University of Hong Kong, Queen Mary Hospital, Hong Kong, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce D.","family":"Given","sequence":"additional","affiliation":[{"name":"Arrowhead Pharmaceuticals, 502 South Rosa Road, Madison, WI 53719, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Wooddell, C.I., Yuen, M.F., Chan, H.L., Gish, R.G., Locarnini, S.A., Chavez, D., Ferrari, C., Given, B.D., Hamilton, J., and Kanner, S.B. (2017). RNAi-based treatment of chronically infected patients and chimpanzees reveals that integrated hepatitis B virus DNA is a source of HBsAg. Sci. Transl. Med., 9.","DOI":"10.1126\/scitranslmed.aan0241"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Tu, T., Budzinska, M.A., Shackel, N.A., and Urban, S. (2017). HBV DNA Integration: Molecular Mechanisms and Clinical Implications. Viruses, 9.","DOI":"10.3390\/v9040075"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1136\/gutjnl-2014-308989","article-title":"Serum viral duplex-linear DNA proportion increases with the progression of liver disease in patients infected with HBV","volume":"65","author":"Zhao","year":"2016","journal-title":"Gut"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"a021360","DOI":"10.1101\/cshperspect.a021360","article-title":"Origins and Evolution of Hepatitis B Virus and Hepatitis D Virus","volume":"6","author":"Littlejohn","year":"2016","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1754","DOI":"10.1136\/gutjnl-2011-301073","article-title":"Innate and adaptive immune responses in chronic hepatitis B virus infections: Towards restoration of immune control of viral infection","volume":"61","author":"Bertoletti","year":"2012","journal-title":"Gut"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1136\/gutjnl-2018-316644","article-title":"Phenotype and function of HBV-specific T cells is determined by the targeted epitope in addition to the stage of infection","volume":"68","author":"Hoogeveen","year":"2019","journal-title":"Gut"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1136\/gutjnl-2018-316641","article-title":"Phenotypic and functional differences of HBV core-specific versus HBV polymerase-specific CD8+ T cells in chronically HBV-infected patients with low viral load","volume":"68","author":"Schuch","year":"2019","journal-title":"Gut"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.jhep.2019.07.015","article-title":"Comparative characterization of B cells specific for HBV nucleocapsid and envelope proteins in patients with chronic hepatitis B","volume":"72","author":"Salimzadeh","year":"2020","journal-title":"J. Hepatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4573","DOI":"10.1172\/JCI121957","article-title":"PD-1 blockade partially recovers dysfunctional virus-specific B cells in chronic hepatitis B infection","volume":"128","author":"Salimzadeh","year":"2018","journal-title":"J. Clin. Investig."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4588","DOI":"10.1172\/JCI121960","article-title":"Circulating and intrahepatic antiviral B cells are defective in hepatitis B","volume":"128","author":"Burton","year":"2018","journal-title":"J. Clin. Investig."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"104816","DOI":"10.1016\/j.antiviral.2020.104816","article-title":"Mechanisms of HBV immune evasion","volume":"179","author":"Kuipery","year":"2020","journal-title":"Antivir. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1055\/s-0039-1685518","article-title":"Hepatitis B Virus-Hepatocyte Interactions and Innate Immune Responses: Experimental Models and Molecular Mechanisms","volume":"39","author":"Thomas","year":"2019","journal-title":"Semin. Liver Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1038\/nm.3856","article-title":"Metabolic regulation of hepatitis B immunopathology by myeloid-derived suppressor cells","volume":"21","author":"Pallett","year":"2015","journal-title":"Nat. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1002\/hep.1840020507","article-title":"Geographic distribution of HBsAg carriers in China","volume":"2","author":"Beasley","year":"1982","journal-title":"Hepatology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"a024935","DOI":"10.1101\/cshperspect.a024935","article-title":"Viral hepatitis B: Clinical and epidemiological characteristics","volume":"4","author":"Burns","year":"2014","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1001\/jama.295.1.65","article-title":"Risk of hepatocellular carcinoma across a biological gradient of serum hepatitis B virus DNA level","volume":"295","author":"Chen","year":"2006","journal-title":"JAMA"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1016\/j.cgh.2007.06.015","article-title":"Risk and predictors of mortality associated with chronic hepatitis B infection","volume":"5","author":"Iloeje","year":"2007","journal-title":"Clin. Gastroenterol. Hepatol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"18035","DOI":"10.1038\/nrdp.2018.35","article-title":"Hepatitis B virus infection","volume":"4","author":"Yuen","year":"2018","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1007\/s10620-014-3486-7","article-title":"Seven-year efficacy and safety of treatment with tenofovir disoproxil fumarate for chronic hepatitis B virus infection","volume":"60","author":"Buti","year":"2015","journal-title":"Dig. Dis. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1016\/j.jhep.2019.11.003","article-title":"Guidance for design and endpoints of clinical trials in chronic hepatitis B\u2014Report from the 2019 EASL-AASLD HBV Treatment Endpoints Conference","volume":"72","author":"Cornberg","year":"2020","journal-title":"J. Hepatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1038\/s41392-020-0207-x","article-title":"Therapeutic siRNA: State of the art","volume":"5","author":"Hu","year":"2020","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Yuen, M.F., Wong, D.K., Schluep, T., Lai, C.L., Ferrari, C., Locarnini, S., Lo, R.C., Gish, R.G., Hamilton, J., and Wooddell, C.I. (2021). Long-term serological, virological and histological responses to RNA inhibition by ARC-520 in Chinese chronic hepatitis B patients on entecavir treatment. Gut.","DOI":"10.1136\/gutjnl-2020-323445"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1038\/mt.2013.31","article-title":"Hepatocyte-targeted RNAi therapeutics for the treatment of chronic hepatitis B virus infection","volume":"21","author":"Wooddell","year":"2013","journal-title":"Mol. Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4284","DOI":"10.1128\/jvi.65.8.4284-4291.1991","article-title":"Diversity of hepatitis B virus X gene-related transcripts in hepatocellular carcinoma: A novel polyadenylation site on viral DNA","volume":"65","author":"Hilger","year":"1991","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Wieland, S.F. (2015). The chimpanzee model for hepatitis B virus infection. Cold Spring Harb. Perspect. Med., 5.","DOI":"10.1101\/cshperspect.a021428"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1092","DOI":"10.1016\/j.jhep.2014.11.031","article-title":"Incidence of hepatocellular carcinoma after HBsAg seroclearance in chronic hepatitis B patients: A need for surveillance","volume":"62","author":"Kim","year":"2015","journal-title":"J. Hepatol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Svicher, V., Salpini, R., Piermatteo, L., Carioti, L., Battisti, A., Colagrossi, L., Scutari, R., Surdo, M., Cacciafesta, V., and Nuccitelli, A. (2020). Whole exome HBV DNA integration is independent of the intrahepatic HBV reservoir in HBeAg-negative chronic hepatitis B. Gut.","DOI":"10.1136\/gutjnl-2020-323300"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"S20","DOI":"10.1016\/S0168-8278(20)30597-3","article-title":"Short-term treatment with RNA interference therapy, JNJ-3989, results in sustained hepatitis B surface antigen supression in patients with chronic hepatitis B receiving nucleos(t)ide analogue treatment","volume":"73","author":"Gane","year":"2020","journal-title":"J. Hepatol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1080\/17512433.2019.1567327","article-title":"Novel developments of hepatitis B: Treatment goals, agents and monitoring tools","volume":"12","author":"Mak","year":"2019","journal-title":"Expert Rev. Clin. Pharm."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1192","DOI":"10.1053\/j.gastro.2008.07.008","article-title":"HBsAg Seroclearance in chronic hepatitis B in Asian patients: Replicative level and risk of hepatocellular carcinoma","volume":"135","author":"Yuen","year":"2008","journal-title":"Gastroenterology"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Pan, X., Zhang, K., Yang, X., Liang, J., Sun, H., Li, X., Zou, Y., Xu, Q., An, G., and Li, G. (2013). Relapse rate and associated-factor of recurrence after stopping NUCs therapy with different prolonged consolidation therapy in HBeAg positive CHB patients. PLoS One, 8.","DOI":"10.1371\/journal.pone.0068568"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6277","DOI":"10.3748\/wjg.v18.i43.6277","article-title":"Durability of viral response after off-treatment in HBeAg positive chronic hepatitis B","volume":"18","author":"Song","year":"2012","journal-title":"World J. Gastroenterol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1016\/j.cgh.2019.07.046","article-title":"Association Between Negative Results From Tests for HBV DNA and RNA and Durability of Response After Discontinuation of Nucles(t)ide Analogue Therapy","volume":"18","author":"Fan","year":"2020","journal-title":"Clin. Gastroenterol. Hepatol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"684","DOI":"10.1053\/j.gastro.2015.11.050","article-title":"Hepatitis B Virus--Specific and Global T-Cell Dysfunction in Chronic Hepatitis B","volume":"150","author":"Park","year":"2016","journal-title":"Gastroenterology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1053\/j.gastro.2012.06.009","article-title":"Preserved T-cell function in children and young adults with immune-tolerant chronic hepatitis B","volume":"143","author":"Kennedy","year":"2012","journal-title":"Gastroenterology"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Heim, K., Binder, B., Wieland, D., Hensel, N., Llewellyn-Lacey, S., Gostick, E., Price, D.A., Emmerich, F., and Vingerhoet, H. (2020). TOX defines the degree of CD8+ T cell dysfunction in distinct phases of chronic HBV infection. Gut.","DOI":"10.1136\/gutjnl-2020-322404"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"830","DOI":"10.1016\/j.jhep.2012.05.015","article-title":"Naive CD8 T cell activation by liver bone marrow-derived cells leads to a \"neglected\" IL-2low Bimhigh phenotype, poor CTL function and cell death","volume":"57","author":"Holz","year":"2012","journal-title":"J. Hepatol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"701","DOI":"10.1172\/JCI200421593","article-title":"The site of primary T cell activation is a determinant of the balance between intrahepatic tolerance and immunity","volume":"114","author":"Bowen","year":"2004","journal-title":"J. Clin. Investig."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Isogawa, M., Chung, J., Murata, Y., Kakimi, K., and Chisari, F.V. (2013). CD40 activation rescues antiviral CD8(+) T cells from PD-1-mediated exhaustion. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003490"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1038\/s41586-019-1620-6","article-title":"Dynamics and genomic landscape of CD8(+) T cells undergoing hepatic priming","volume":"574","author":"Benechet","year":"2019","journal-title":"Nature"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2940","DOI":"10.1128\/JVI.02415-06","article-title":"The level of viral antigen presented by hepatocytes influences CD8 T-cell function","volume":"81","author":"Gehring","year":"2007","journal-title":"J. Virol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1762","DOI":"10.1053\/j.gastro.2020.01.032","article-title":"Knockdown of Virus Antigen Expression Increases Therapeutic Vaccine Efficacy in High-Titer Hepatitis B Virus Carrier Mice","volume":"158","author":"Michler","year":"2020","journal-title":"Gastroenterology"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Balagopal, A., Grudda, T., Ribeiro, R.M., Saad, Y.S., Hwang, H.S., Quinn, J., Murphy, M., Ward, K., Sterling, R.K., and Zhang, Y. (2020). Single hepatocytes show persistence and transcriptional inactivity of hepatitis B. JCI Insight, 5.","DOI":"10.1172\/jci.insight.140584"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.1038\/sj.mt.6300177","article-title":"Histone modifications are associated with the persistence or silencing of vector-mediated transgene expression in vivo","volume":"15","author":"Riu","year":"2007","journal-title":"Mol. Ther."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1006\/jmbi.2000.4481","article-title":"Structural organization of the hepatitis B virus minichromosome","volume":"307","author":"Bock","year":"2001","journal-title":"J. Mol. Biol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1016\/j.jhep.2020.09.019","article-title":"Dicoumarol, an NQO1 inhibitor, blocks cccDNA transcription by promoting degradation of HBx","volume":"74","author":"Cheng","year":"2021","journal-title":"J. Hepatol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1002\/jcp.21172","article-title":"Liver regeneration","volume":"213","author":"Michalopoulos","year":"2007","journal-title":"J. Cell Physiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/S0925-4773(02)00338-6","article-title":"The role of hepatocytes and oval cells in liver regeneration and repopulation","volume":"120","author":"Fausto","year":"2003","journal-title":"Mech. Dev."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1097\/MPG.0b013e3181e7ff55","article-title":"Hepatic progenitor cell proliferation and liver injury in alpha-1-antitrypsin deficiency","volume":"51","author":"Brunt","year":"2010","journal-title":"J. Pediatr. Gastroenterol. Nutr."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1016\/j.ajpath.2014.02.020","article-title":"The liver is a peculiar organ when it comes to stem cells","volume":"184","author":"Michalopoulos","year":"2014","journal-title":"Am. J. Pathol."},{"key":"ref_51","first-page":"1A","article-title":"Tenfovir disoproxil fumarate treatment reduces the number of transcriptionally active viral integrations in chronically infected HBV patients","volume":"72","author":"Hsu","year":"2020","journal-title":"Hepatology"},{"key":"ref_52","first-page":"1A","article-title":"Long term nucleos(t)ide analog therapy reduced the extent of HBV DNA integration in chronic hepatitis B patients","volume":"72","author":"Chow","year":"2020","journal-title":"Hepatology"}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/4\/581\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T14:10:29Z","timestamp":1760364629000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/13\/4\/581"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,30]]},"references-count":52,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2021,4]]}},"alternative-id":["v13040581"],"URL":"https:\/\/doi.org\/10.3390\/v13040581","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,30]]}}}