| Project/Area Number |
20K22771
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Multi-year Fund |
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| Review Section |
0803:Pathology, infection/immunology, and related fields
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| Research Institution | Jichi Medical University |
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Principal Investigator |
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| Project Period (FY) |
2020-09-11 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | hepatitis E virus / bioluminescent / reporter virus / nanoKAZ / hypervariable region / non-enveloped HEV / quasi-enveloped HEV / drug screening / Hepatitis E virus / Bioluminescence / Drug-screening / Cell-culture / Drug screening |
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| Outline of Research at the Start |
Certain hepatitis E cases including fulminant or chronic cases require antiviral treatment. However, specific anti-hepatitis E virus (HEV) drug is currently unavailable. In this study, the applicant will try to identify novel candidates with anti-HEV activity via comprehensive drug screening to FDA-approved drug library and small compound library using infectious HEV harboring nanoKAZ reporter gene. Effectiveness and inhibitory mechanism of selected drugs will be analyzed in vitro. The results will provide new insights into the HEV life cycle for the development of novel anti-HEV drugs.
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| Outline of Final Research Achievements |
A system consisting of recombinant infectious hepatitis E virus (HEV) harboring a small luciferase gene (nanoKAZ) in ORF1, was developed in this study. It replicated efficiently in cultured cells, was genetically stable, and had morphological characteristics similar to those of the parental virus. Both membrane-associated (eHEV-nanoKAZ) and membrane-unassociated (neHEV-nanoKAZ) particles were infectious. The system was successfully applied in a screening to search for novel anti-HEV drugs. This screening system was able to cover the inhibitor of HEV entry and HEV RNA replication. In the screening, four drugs were identified and confirmed to be effective in cultured cells. Two drugs (azithromycin and ritonavir) strongly inhibited HEV production in culture supernatants, and intracellular expression of ORF2 protein, and may therefore be candidate novel anti-HEV drugs. The results of this study provide evidence supporting the use of this system in more variable HEV studies.
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| Academic Significance and Societal Importance of the Research Achievements |
新たに開発したHEV-nanoKAZを利用することにより、これまで同定することのできなかった感染初期過程を阻害する薬剤のスクリーニングが可能となった。今後、さまざまな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、膜に覆われたHEVと膜に覆われていないHEVが利用する感染受容体は同定されておらず、細胞内侵入機構についても不明な点が多く残されている。HEV-nanoKAZシステムは、このようなHEVのライフサイクルの研究へとさらに応用が可能である。
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