Inhibition of HBV Transcription From cccDNA With Nitazoxanide by Targeting the HBx–DDB1 Interaction
| Autor: | Rei Ishibashi, Motoyuki Otsuka, Eri Tanaka, Motoko Ohno, Mari Yamagami, Takahiro Kishikawa, Takahiro Seimiya, Kazuhiko Koike, Tatsunori Suzuki, Kazuma Sekiba |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
DDB1 damage-specific DNA-binding protein 1 viruses mcHBV minicircle hepatitis B virus DMSO dimethyl sulfoxide Flag-HBx Flag-tagged hepatitis B virus regulatory protein X medicine.disease_cause Cluc Cypridina luciferase 0302 clinical medicine Transcription (biology) LgBit Large Bit PSAD plasmid-safe DNase HBx hepatitis B virus regulatory protein X Original Research Smc5/6 structural maintenance of chromosomes 5/6 DMEM Dulbecco's modified Eagle's medium HBs hepatitis B surface antibody Gastroenterology virus diseases IFNα interferon alfa IP immunoprecipitation Nitazoxanide cccDNA Nitro Compounds mRNA messenger RNA HBx qPCR quantitative polymerase chain reaction 030211 gastroenterology & hepatology cccDNA covalently closed circular DNA medicine.drug Hepatitis B virus SDS sodium dodecyl sulfate Viral protein Biology Minicircle 03 medical and health sciences DDB1 medicine lcsh:RC799-869 Primary Hepatocyte Infection ddPCR droplet digital polymerase chain reaction Gluc Gaussia luciferase Drug Screening Hepatology pgRNA pregenomic RNA CMV cytomegalovirus FDA Food and Drug Administration Virology digestive system diseases Thiazoles HBV hepatitis B virus 030104 developmental biology NTZ nitazoxianide Viral replication SmBit Small Bit lcsh:Diseases of the digestive system. Gastroenterology |
| Zdroj: | Cellular and Molecular Gastroenterology and Hepatology, Vol 7, Iss 2, Pp 297-312 (2019) Cellular and Molecular Gastroenterology and Hepatology |
| ISSN: | 2352-345X |
| Popis: | Background & Aims Hepatitis B virus (HBV) infection is a major health concern worldwide. Although currently used nucleos(t)ide analogs efficiently inhibit viral replication, viral proteins transcribed from the episomal viral covalently closed circular DNA (cccDNA) minichromosome continue to be expressed long-term. Because high viral RNA or antigen loads may play a biological role during this chronicity, the elimination of viral products is an ultimate goal of HBV treatment. HBV regulatory protein X (HBx) was recently found to promote transcription of cccDNA with degradation of Smc5/6 through the interaction of HBx with the host protein DDB1. Here, this protein–protein interaction was considered as a new molecular target of HBV treatment. Methods To identify candidate compounds that target the HBx–DDB1 interaction, a newly constructed split luciferase assay system was applied to comprehensive compound screening. The effects of the identified compounds on HBV transcription and cccDNA maintenance were determined using HBV minicircle DNA, which mimics HBV cccDNA, and the natural HBV infection model of human primary hepatocytes. Results We show that nitazoxanide (NTZ), a thiazolide anti-infective agent that has been approved by the FDA for protozoan enteritis, efficiently inhibits the HBx–DDB1 protein interaction. NTZ significantly restores Smc5 protein levels and suppresses viral transcription and viral protein production in the HBV minicircle system and in human primary hepatocytes naturally infected with HBV. Conclusions These results indicate that NTZ, which targets an HBV-related viral–host protein interaction, may be a promising new therapeutic agent and a step toward a functional HBV cure. Graphical abstract |
| Databáze: | OpenAIRE |
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