| Project/Area Number |
22K15478
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 49060:Virology-related
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| Research Institution | Jichi Medical University |
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Principal Investigator |
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| Project Period (FY) |
2022-04-01 – 2024-03-31
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| Project Status |
Completed (Fiscal Year 2023)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2023: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2022: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | Hepatitis E virus / Host cellular factor / Anti-HEV drug / Microarray / Cell culture / Replication efficiency / Reporter virus / Drug screening / Cellular permissiveness / Host factor / Small compound screening / Virus replication |
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| Outline of Research at the Start |
Patients with acute fulminant or chronic hepatitis E require antiviral drug. However, specific drug is currently unavailable. Subclones of PLC/PRF/5 cells showed variable permissiveness to hepatitis E virus (HEV) replication. Results of RNA microarray analysis of selected subclones will be used to investigate cellular factors and pathways responsible for the determination of cell permissiveness to HEV replication, and the small compounds which can act as their specific inhibitors. The results of this study will provide valuable data for the development of novel and specific anti-HEV drugs.
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| Outline of Final Research Achievements |
Subclones of a single PLC/PRF/5 cell line demonstrated up to 10,000-folds difference in the permissiveness to hepatitis E virus (HEV) replication. Based on the results of RNA microarray analysis of highly permissive and poorly permissive subclones, several genes were silenced by using small interfering RNA (siRNA) followed by screening using eHEV-nanoKAZ and evaluation in cell culture. Among them, possible genes involved in cellular permissiveness to HEV were identified. One hit gene was used as the target for screening on a small compound library by three distinct HEV reporter systems. Among the hit compounds, one with strongest inhibition on luciferase activity was further evaluated in cultured cells. It exhibited moderate inhibition on HEV growth in an evaluation in cell culture system using a wild-type HEV-3 strain. The results of this study will enhance the insights on HEV molecular aspects and will be useful for the development of novel and specific anti-HEV drug.
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| Academic Significance and Societal Importance of the Research Achievements |
新たに同定されたHEV感染の許容性に関与する宿主因子は、新規抗HEV薬開発の標的と成り得ることが示唆された。今後、さまざまな関連ライブラリを用いたスクリーニングを実施することで、より効果の高い抗HEV候補薬が同定されることが期待される。また、HEVの複製機構に関しては未だ不明な点が多く残されている。今回同定された宿主因子は、HEVの感染や複製の分子機構を理解する上でも重要な知見である。
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