2016 Fiscal Year Annual Research Report
核外移行機序を標的とした新規インフルエンザ薬の同定
| Project/Area Number |
15F15416
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
間 陽子 国立研究開発法人理化学研究所, 分子ウイルス学特別研究ユニット, ユニットリーダー (50182994)
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| Co-Investigator(Kenkyū-buntansha) |
CHUTIWITOONCHAI NOPPORN 国立研究開発法人理化学研究所, 分子ウイルス学特別研究ユニット, 外国人特別研究員
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| Project Period (FY) |
2015-10-09 – 2018-03-31
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| Keywords | Influenza A virus / nucleoprotein (NP) / NXT1 / nuclear export / inhibitor / CRM1 |
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| Outline of Annual Research Achievements |
1) We reported NXT1 as a new identified host factor that promotes nuclear export of influenza nucleoprotein (NP) via cellular CRM1 pathway (Chutiwitoonchai et al., Viruses 2016). In detail, NXT1 binds at the C-terminal region of NP and forms a complex with CRM1 for promoting NP nuclear export which is an important mechanism for viral assembly and replication.
2) We reported that inhibition of NP nuclear export function via the NP-CRM1 interaction is a promising therapeutic strategy against influenza infection (Chutiwitoonchai et al., Virology 2017). In detail, blocking NP-CRM1 binding by a small compound broadly inhibited replication of several influenza A subtypes.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
1) Identification of NXT1 as a new host factor that promotes nuclear export function of influenza NP. We demonstrated that knockdown of NXT1 resulted in nuclear accumulation of NP including viral genomic RNA and decreased viral replication kinetic while overexpression of NXT1 promoted nuclear export of NP. We also showed that NXT1 bound at the C-terminal region of NP and formed a complex together with CRM1 for promoting NP nuclear export.
2) Investigation of an inhibitory mechanism and broad range effect of a small compound that inhibits nuclear export function of influenza NP. This study demonstrated targeting NP nuclear export function at the NP-CRM1 interaction by a small compound, efficiently inhibited viral replication. In addition, this inhibitory mechanism also affected replication of broad range influenza A subtypes.
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| Strategy for Future Research Activity |
1) To identify a new small compound analog with improving antiviral activity. A similar chemical structure of the small compound will be selected and purchased from available database for testing antiviral activity against A/WSN/33 replication by plaque assay and cytotoxicity by WST-1 assay. Then, the inhibitory effect will be validated by immunofluorescent stain of NP and pull-down assay of NP/CRM1.
2) To evaluate an antiviral effect of a small compound against virus that uses cellular CRM1 nuclear export pathway e.g., HIV-1. A broad range inhibitory effect of a small compound will be tested against HIV-1 replication by treating NL4-3 infected CEM cells with a small compound. Viral replication efficiency will be determined by p24 Gag ELISA and cytotoxicity will be tested by WST-1 assay. The mechanism of inhibition will be confirmed by immunofluorescent stain of viral Rev and pull-down assay of Rev/CRM1.
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Research Products
(4 results)
