Hepatitis B virus sensitivity to interferon‐α in hepatocytes is more associated with cellular interferon response than with viral genotype

Shen, Fang; Li, Yaming; Wang, Yang; Sozzi, Vitina; Revill, Peter; Liu, Jiangxia; Gao, Lu; Yang, Guang; Lu, Mengji; Sutter, Kathrin; Dittmer, Ulf; Chen, Jieliang; Yuan, Zhenghong

doi:10.1002/hep.29609

Cited by 57 publications

(60 citation statements)

References 50 publications

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“…Previous study revealed that the innate immune pathway was not induced in CHB patients’ liver, which could be reactivated following stimulation of Toll‐like receptor (TLR)3 by poly I:C or virus . Yuan et al also found that host factors played an important part in antiviral immunity, whereas HBV sensitivity to IFNα was more related to activation of cellular IFN pathway than the viral genotype . Because the analysis of GEO data guided us to focus on host immunity, we applied PCR array to investigate factors concerning the innate immunity against virus.…”

Section: Discussionmentioning

confidence: 99%

“…(22) Yuan et al also found that host factors played an important part in antiviral immunity, whereas HBV sensitivity to IFNα was more related to activation of cellular IFN pathway than the viral genotype. (23) Because the analysis of GEO data guided us to focus on host immunity, we applied PCR array to investigate factors concerning the innate immunity against virus. Consistently, our study demonstrated lower baseline level of IFNα mRNA in PBMCs of CHB patients with suboptimal response to IFNα treatment, whereas there was an increased baseline level of the host immune regulator, IFITM2, indicating that an impaired immune pathway may be responsible for IFNα inefficiency (Fig.…”

Section: Discussionmentioning

confidence: 99%

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Exosomal Interferon‐Induced Transmembrane Protein 2 Transmitted to Dendritic Cells Inhibits Interferon Alpha Pathway Activation and Blocks Anti–Hepatitis B Virus Efficacy of Exogenous Interferon Alpha

Shi

et al. 2019

Hepatology

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The negative regulators in the interferon (IFN) signaling pathway inhibit intrahepatic immune response, resulting in suboptimal therapeutic response to IFNα treatment in chronic hepatitis B (CHB) patients. Identifying the key negative factors and elucidating the regulating mechanism are essential for improving anti‐HBV (hepatitis B virus) efficacy of IFNα. From the Gene Expression Omnibus (GEO) database, we downloaded and analyzed gene expression profiles of CHB patients with different responses to IFNα (GSE54747), and found that innate immune status was associated with the IFNα‐based therapeutic response in CHB patients. Through PCR array, we found higher baseline level of IFN‐induced transmembrane protein 2 (IFITM2) mRNA and lower baseline level of IFNα mRNA in peripheral blood mononuclear cells (PBMCs) of CHB patients with suboptimal response to IFNα treatment. Increased IFITM2 protein was also found in the serum of IFNα nonresponsive patients. With further experiments, we found that overexpressing IFITM2 in Huh7 cells suppressed endogenous IFNα synthesis by inhibiting phosphorylation of extracellular signal–regulated kinase (ERK), TANK‐binding kinase 1 (TBK1), and interferon regulatory factor 3 (IRF3); knocking out IFITM2 enhanced activation of the endogenous IFNα synthesis pathway, exhibiting better inhibition on HBV replication. We also found that IFITM2 protein was shuttled by exosomes to dendritic cells (DCs), the main source of endogenous IFNα. Exosome‐mediated transport of IFITM2 inhibited synthesis of endogenous IFNα in DCs whereas the inhibitory effect was abolished when IFITM2 was knocked out. Furthermore, we demonstrated that both palmitoylation inhibitor and mutation on 70/71 sites of IFITM2 protein influenced its incorporation into exosomes. Mutated IFITM2 protein increased the effect of IFNα against HBV. Conclusion: Exosome‐mediated transport of IFITM2 to DCs inhibits IFNα pathway activation and blocks anti‐HBV efficacy of exogenous IFNα. The findings provide an explanation to the suboptimal response of CHB patients to IFNα treatment.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Exosomal Interferon‐Induced Transmembrane Protein 2 Transmitted to Dendritic Cells Inhibits Interferon Alpha Pathway Activation and Blocks Anti–Hepatitis B Virus Efficacy of Exogenous Interferon Alpha

Shi

et al. 2019

Hepatology

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“…Proliferating HepaRG cells were purchased from Biopredic International (Rennes, France) and were amplified and differentiated according to the manufacturer's protocol. HepG2-NTCP (kindly provided by Yi Ni and Stephan Urban) and primary human hepatocytes (PHH; purchased from Bioreclamation IVT) were cultured as described previously (41). The human hepatomaderived cell line HepG2 (purchased from ATCC) was cultured in Dulbecco's modified Eagle medium (DMEM)/F12 supplemented with 10% fetal bovine serum (Invitrogen), 2 mM L-glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin at 37°C under humidified air that contained 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

“…HBV infection. HBV was prepared from the HepAD38 cells (43), and the infection was performed with 150 GE per cell in the presence of 4% (vol/vol) polyethylene glycol 8000 (PEG 8000; Sigma-Aldrich) at 37°C for 24 h, as we described previously (41). The cells were washed and maintained in medium with a medium change every 2 to 3 days.…”

Section: Methodsmentioning

confidence: 99%

Identification of Retinoic Acid Receptor Agonists as Potent Hepatitis B Virus Inhibitors via a Drug Repurposing Screen

Wang

Shen

et al. 2018

Antimicrob Agents Chemother

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Currently available therapies for chronic hepatitis B virus (HBV) infection can efficiently reduce viremia but induce hepatitis B surface antigen (HBsAg) loss in very few patients; also, these therapies do not greatly affect the viral covalently closed circular DNA (cccDNA). To discover new agents with complementary anti-HBV effects, we performed a drug repurposing screen of 1,018 Food and Drug Administration (FDA)-approved compounds using HBV-infected primary human hepatocytes (PHH). Several compounds belonging to the family of retinoic acid receptor (RAR) agonists were identified that reduced HBsAg levels in a dose-dependent manner without significant cytotoxicity. Among them, tazarotene exhibited the most potent anti-HBV effect, with a half-maximal inhibitory concentration (IC) for HBsAg of less than 30 nM in PHH. The inhibitory effect was also observed in HBV-infected differentiated HepaRG (dHepaRG) models, but not in HepG2.215 cells, and HBV genotypes A to D were similarly inhibited. Tazarotene was further demonstrated to repress HBV cccDNA transcription, as determined by the levels of HBV cccDNA and RNAs and the activation of HBV promoters. Moreover, RNA sequence analysis showed that tazarotene did not induce an interferon response but altered the expression of a number of genes associated with RAR and metabolic pathways. Inhibition of RARβ, but not RARα, by a specific antagonist significantly attenuated the anti-HBV activity of tazarotene, suggesting that tazarotene inhibits HBV in part through RARβ. Finally, a synergistic effect of tazarotene and entecavir on HBV DNA levels was observed. Therefore, RAR agonists as represented by tazarotene were identified as potential novel anti-HBV agents.

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“…Interferon (IFN)-α, a type I IFN, modulates the immune response to HBV infection by activating multiple IFNstimulated genes (ISGs), which act to inhibit HBV replication by affecting multiple steps of the viral life cycle and inducing the decay of established HBV cccDNA. 3,7 IFN-γ, a type II IFN, is produced by intrahepatic cytotoxic T lymphocytes, and has non-cytolytic antiviral potential. 8,9 Using a transgenic mouse model of chronic hepatitis B, we showed that CD8 + cytotoxic T lymphocytes are primarily responsible for the induction of chronic liver disease.…”

Section: Introductionmentioning

confidence: 99%

Molecular signature of hepatitis B virus regulation by interferon‐γ in primary human hepatocytes

Nosaka

Naito

Matsuda

et al. 2019

Hepatology Research

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Aim A complete cure for chronic hepatitis B virus (HBV) infection requires elimination of covalently closed circular DNA; however, this remains to be clinically achieved. Interferon (IFN)‐γ, a type II IFN, is produced by intrahepatic cytotoxic T lymphocytes and has non‐cytolytic antiviral potential. However, the mechanism by which IFN‐γ regulates HBV infection has not been fully elucidated. Thus, we developed an in vitro HBV infection assay system and analyzed the molecular signature of HBV regulation by IFN‐γ. Methods The in vitro HBV infection assay system was established in primary human hepatocytes infected with HBV derived from the plasmid containing 1.3‐mer HBV genome, and treated with IFN‐γ. The antiviral effects and signaling pathways of IFN‐γ were examined using microarray, and assessed by siRNA knockdown experiments of the related genes. Results IFN‐γ treatment suppressed both HBV propagation and transcription as efficiently as IFN‐α. Microarray analysis showed that IFN‐γ stimulation induced the activation of both IFN‐γ and IFN‐α signaling, regulating HBV covalently closed circular DNA. HBV production was decreased by IFN‐γ through Janus kinase/signal transducer and activator of transcription signaling and interferon‐stimulated genes, such as 2′‐5′‐oligoadenylate synthase 2 and apolipoprotein B mRNA editing enzyme catalytic subunit 3G. Conclusions IFN‐γ can suppress HBV propagation and transcription in hepatocytes by activating specific intracellular signaling pathways in hepatocytes, and suggests the future application of these particular signaling pathways or genes for the complete elimination of HBV.

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Hepatitis B virus sensitivity to interferon‐α in hepatocytes is more associated with cellular interferon response than with viral genotype

Cited by 57 publications

References 50 publications

Exosomal Interferon‐Induced Transmembrane Protein 2 Transmitted to Dendritic Cells Inhibits Interferon Alpha Pathway Activation and Blocks Anti–Hepatitis B Virus Efficacy of Exogenous Interferon Alpha

Exosomal Interferon‐Induced Transmembrane Protein 2 Transmitted to Dendritic Cells Inhibits Interferon Alpha Pathway Activation and Blocks Anti–Hepatitis B Virus Efficacy of Exogenous Interferon Alpha

Identification of Retinoic Acid Receptor Agonists as Potent Hepatitis B Virus Inhibitors via a Drug Repurposing Screen

Molecular signature of hepatitis B virus regulation by interferon‐γ in primary human hepatocytes

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