Highlights
- •we define the HDV-induced signature transcriptomic modulations mainly composed of ISGs
- •ISGs are upregulated upon HDV infection but not in cells only expressing HDAg alone
- •HDV interferes with HBV through both IFN-dependent and IFN-independent mechanisms
- •HDV infection and HDAg expression decrease the levels of nascent HBV RNAs and accelerated the decay of HBV RNA
- •HDAg are associated with HBV RNAs
Abstract
Background & Aims
Chronic co-infection with hepatitis B and D viruses (HBV and HDV) leads to the most
aggressive form of chronic viral hepatitis. Here, we aimed at elucidating the molecular
mechanisms leading to the interference of HDV on HBV observed in most co-infected
patients.
Methods
Patient liver tissues, primary human hepatocytes, HepaRG cells and human liver chimeric
mice were used to analyze the effect of HDV on HBV using virological and RNA-seq analyses,
as well as RNA synthesis, stability and association assays.
Results
Transcriptomic analyses in cell culture and mouse models of co-infection allowed to
define the HDV-induced signature mainly composed of interferon (IFN)-stimulated genes
(ISGs). We also provide evidence that ISGs are upregulated in HDV-HBV chronically
infected patients but not in cells only expressing the HDV antigens (HDAg). Inhibition
of the hepatocyte IFN response partially rescued the levels of HBV parameters. We
observed less HBV RNAs synthesis upon HDV infection or HDV protein expression. Additionally,
HDV infection or expression of HDAg alone specifically accelerated the decay of HBV
RNAs and HDAg are associated with HBV RNAs. On the contrary, HDAg expression did not
affect other viruses such as hepatitis C virus (HCV) or severe acute respiratory syndrome
coronavirus 2 (SARS-CoV-2).
Conclusions
Our data indicate that HDV interferes with HBV through both IFN-dependent and IFN-independent
mechanisms. Specifically, we uncover a new viral interference mechanism in which proteins
of a satellite virus affect RNA production of its helper virus. Exploiting these finding
could pave the way to the development of new therapeutic strategies against HBV.
Impact and implications
Although, the molecular mechanisms remained unexplored, it was known for long that
despite its dependency, HDV decreased HBV viremia in patients. Here, using in vitro
and in vivo models, we showed that HDV interferes with HBV through both IFN-dependent
and IFN-independent mechanisms affecting HBV RNAs metabolism and we defined the HDV-induced
modulation signature. The mechanisms we uncovered could pave the way to the development
of new therapeutic strategies against HBV by mimicking and/or increasing the effect
of HDAg on HBV RNAs and the HDV-induced modulation signature may allow to the draw
correlation withthe responsiveness to IFN alpha treatment and thereby ultimately help
in the management of HBV/HDV co-infected patients.
Graphical abstract
Graphical Abstract
KeyWords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Journal of HepatologyAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
Author names in bold designate shared co-first authorship.
- The covalently closed duplex form of the hepadnavirus genome exists in situ as a heterogeneous population of viral minichromosomes.J Virol. 1995; 69: 3350-3357
- Hepatitis B virus genome is organized into nucleosomes in the nucleus of the infected cell.Virus Genes. 1994; 8: 215-229
- Structural organization of the hepatitis B virus minichromosome.Journal of molecular biology. 2001; 307: 183-196
- Nuclear HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function.Proc Natl Acad Sci U S A. 2009; 106: 19975-19979
- Hepatitis B virus X protein is essential to initiate and maintain virus replication after infection.J Hepatol. 2011; 55: 996-1003
- Hepatitis B virus replication is regulated by the acetylation status of hepatitis B virus cccDNA-bound H3 and H4 histones.Gastroenterology. 2006; 130: 823-837
- Methyltransferase PRMT1 is a binding partner of HBx and a negative regulator of hepatitis B virus transcription.J Virol. 2013; 87: 4360-4371
- IFN-alpha inhibits HBV transcription and replication in cell culture and in humanized mice by targeting the epigenetic regulation of the nuclear cccDNA minichromosome.J Clin Invest. 2012; 122: 529-537
- Transmission of the hepatitis B virus-associated delta antigen to chimpanzees.J Infect Dis. 1980; 141: 590-602
- Origin of hepatitis delta virus mRNA.J Virol. 2000; 74: 7204-7210
- Hepatitis delta virus genome replication: a polyadenylated mRNA for delta antigen.J Virol. 1990; 64: 3192-3198
- Current knowledge on Hepatitis Delta Virus replication.Antiviral Res. 2020; 179104812
- Liver disease activity and hepatitis B virus replication in chronic delta antigen-positive hepatitis B virus carriers.Hepatology. 1985; 5: 544-547
- Hepatitis delta virus (HDV) and woodchuck hepatitis virus (WHV) nucleic acids in tissues of HDV-infected chronic WHV carrier woodchucks.J Virol. 1989; 63: 1612-1618
- Humanized chimeric uPA mouse model for the study of hepatitis B and D virus interactions and preclinical drug evaluation.Hepatology. 2012; 55: 685-694
- Replicative and transcriptional activities of hepatitis B virus in patients coinfected with hepatitis B and hepatitis delta viruses.J Virol. 2011; 85: 432-439
- Single (B or C), dual (BC or BD) and triple (BCD) viral hepatitis in HIV-infected patients in Madrid, Spain.Aids. 2005; 19: 1361-1365
- Smouldering hepatitis B virus replication in patients with chronic liver disease and hepatitis delta virus superinfection.J Hepatol. 1991; 12: 64-69
- Hepatitis D virus (delta agent) superinfection in an endemic area of hepatitis B infection: immunopathologic and serologic findings.Scand J Infect Dis. 1987; 19: 173-177
- Hepatitis B virus clearance from serum and liver after acute hepatitis delta virus superinfection in chronic HBsAg carriers.J Med Virol. 1990; 31: 284-290
- Interaction between HDV and HBV infection in HBsAg-chronic carriers.Infection. 1991; 19: 155-158
- Quantitative longitudinal evaluations of hepatitis delta virus RNA and hepatitis B virus DNA shows a dynamic, complex replicative profile in chronic hepatitis B and D.J Hepatol. 2010; 52: 658-664
- Viral dominance patterns in chronic hepatitis delta determine early response to interferon alpha therapy.J Viral Hepat. 2018; 25: 1384-1394
- HDV RNA replication is associated with HBV repression and interferon-stimulated genes induction in super-infected hepatocytes.Antiviral Res. 2016; 136: 19-31
- Production of hepatitis delta virus and suppression of helper hepatitis B virus in a human hepatoma cell line.J Virol. 1991; 65: 1099-1104
- Hepatitis delta virus proteins repress hepatitis B virus enhancers and activate the alpha/beta interferon-inducible MxA gene.J Gen Virol. 2009; 90: 2759-2767
- Hepatitis Delta co-infection in humanized mice leads to pronounced induction of innate immune responses in comparison to HBV mono-infection.J Hepatol. 2015; 63: 346-353
- Toll-like receptor 7 agonist GS-9620 induces prolonged inhibition of HBV via a type I interferon-dependent mechanism.J Hepatol. 2018; 68: 922-931
- Inhibition of type I interferon-mediated antiviral action in human glioma cells by the IKK inhibitors BMS-345541 and TPCA-1.Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research. 2012; 32: 368-377
- Hepatitis D virus replication is sensed by MDA5 and induces IFN-beta/lambda responses in hepatocytes.J Hepatol. 2018;
- The large form of hepatitis delta antigen is crucial for assembly of hepatitis delta virus.Proc Natl Acad Sci U S A. 1991; 88: 8490-8494
- The large delta antigen of hepatitis delta virus potently inhibits genomic but not antigenomic RNA synthesis: a mechanism enabling initiation of viral replication.J Virol. 2000; 74: 7375-7380
- PAPD5/7 Are Host Factors That Are Required for Hepatitis B Virus RNA Stabilization.Hepatology. 2018;
- When Hepatitis B Virus Meets Interferons.Front Microbiol. 2018; 9: 1611
- N6-methyladenosine modification of hepatitis B virus RNA differentially regulates the viral life cycle.Proc Natl Acad Sci U S A. 2018; 115: 8829-8834
- Structure and function of HCV IRES domains.Virus Res. 2009; 139: 166-171
- Unpacking Pandora From Its Box: Deciphering the Molecular Basis of the SARS-CoV-2 Coronavirus.Int J Mol Sci. 2020; 22
- Preclinical assessment of antiviral combination therapy in a genetically humanized mouse model for hepatitis delta virus infection.Sci Transl Med. 2018; 10
- Hepatitis D Virus Infection of Mice Expressing Human Sodium Taurocholate Co-transporting Polypeptide.PLoS Pathog. 2015; 11e1004840
- A new HDV mouse model identifies mitochondrial antiviral signaling protein (MAVS) as a key player in IFN-beta induction.J Hepatol. 2017;
- Small hepatitis delta antigen selectively binds to target mRNA in hepatic cells: a potential mechanism by which hepatitis D virus downregulates glutathione S-transferase P1 and induces liver injury and hepatocarcinogenesis.Biochem Cell Biol. 2019; 97: 130-139
- IL28 variation affects expression of interferon stimulated genes and peg-interferon and ribavirin therapy.J Hepatol. 2011; 54: 1094-1101
- Hepatic ISG expression is associated with genetic variation in interleukin 28B and the outcome of IFN therapy for chronic hepatitis C.Gastroenterology. 2010; 139: 499-509
- Hepatitis delta virus ribonucleoproteins shuttle between the nucleus and the cytoplasm.RNA. 2002; 8: 637-646
- Interferon-inducible ribonuclease ISG20 inhibits hepatitis B virus replication through directly binding to the epsilon stem-loop structure of viral RNA.PLoS Pathog. 2017; 13e1006296
- Interplay between Hepatitis D Virus and the Interferon Response.Viruses. 2020; 12
- Tripartite motif-containing 22 inhibits the activity of hepatitis B virus core promoter, which is dependent on nuclear-located RING domain.Hepatology. 2009; 50: 424-433
- Extracellular Vesicles Including Exosomes Regulate Innate Immune Responses to Hepatitis B Virus Infection.Front Immunol. 2016; 7: 335
- Interferon-stimulated gene 20 (ISG20) selectively degrades N6-methyladenosine modified Hepatitis B Virus transcripts.PLoS Pathog. 2020; 16e1008338
- Inducers of the NF-kappaB pathways impair hepatitis delta virus replication and strongly decrease progeny infectivity in vitro.JHEP Rep. 2022; 4100415
- Structural basis of the oligomerization of hepatitis delta antigen.Structure. 1998; 6: 821-830
- Determination of the multimerization state of the hepatitis delta virus antigens in vivo.J Virol. 2003; 77: 10314-10326
- Intrinsic disorder and oligomerization of the hepatitis delta virus antigen.Virology. 2010; 407: 333-340
- Arginine-rich motifs are not required for hepatitis delta virus RNA binding activity of the hepatitis delta antigen.J Virol. 2013; 87: 8665-8674
- A Genome-wide CRISPR Screen Identifies ZCCHC14 as a Host Factor Required for Hepatitis B Surface Antigen Production.Cell reports. 2019; 29: 2970-2978 e2976
- Viral hijacking of the TENT4-ZCCHC14 complex protects viral RNAs via mixed tailing.Nature structural & molecular biology. 2020; 27: 581-588
- Interferon-Dependent and Respiratory Virus-Specific Interference in Dual Infections of Airway Epithelia.Scientific reports. 2020; 1010246
- Inhibition of herpes simplex virus type 1 replication by adeno-associated virus rep proteins depends on their combined DNA-binding and ATPase/helicase activities.J Virol. 2010; 84: 3808-3824
- Adeno-Associated Virus Type 2 Rep68 Can Bind to Consensus Rep-Binding Sites on the Herpes Simplex Virus 1 Genome.J Virol. 2015; 89: 11150-11158
- Envelope proteins derived from naturally integrated hepatitis B virus DNA support assembly and release of infectious hepatitis delta virus particles.J Virol. 2014; 88: 5742-5754
Article info
Publication history
Accepted:
January 10,
2023
Received in revised form:
January 10,
2023
Received:
January 31,
2022
Publication stage
In Press Journal Pre-ProofIdentification
Copyright
© 2023 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
