2020 Fiscal Year Research-status Report
Production of infectious hepatitis E virus (HEV) harboring bioluminescent reporter gene for comprehensive screening of antiviral drugs against HEV and ex vivo evaluation of selected drugs
| Project/Area Number |
20K22771
|
|---|
| Research Institution | Jichi Medical University |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2020-09-11 – 2022-03-31
|
| Keywords | Hepatitis E virus / Bioluminescence / nanoKAZ / Drug-screening / Cell-culture |
|---|
| Outline of Annual Research Achievements |
Hepatitis E virus (HEV) is the causative agent of acute self-limiting, fulminant, or chronic hepatitis. Antiviral treatment is required for certain hepatitis E cases including fulminant or chronic cases. However, specific antiviral drug against HEV is currently unavailable. In order to identify novel candidates with activity against HEV, infectious HEV progenies expressing bioluminescent reporter gene (small luciferase: nanoKAZ) was constructed to be used for comprehensive drug screening to the food and drug administration (FDA)-approved drug library and small compound library. Generation of HEV progenies, the intracellular expression levels of nanoKAZ, the infectivity of the recombinant virus, and the utility of this screening system were confirmed.
Comprehensive drug screening to the FDA-approved drug library revealed four candidate drugs with anti-HEV activity, which are different to the drugs found in our previous screening to the same FDA-approved drug library using HEV replicon expressing Gaussia luciferase. These results suggested that rather than inhibiting HEV replication, the four candidate drugs in the current study act to inhibit different step in HEV life cycle. Furthermore, effectiveness of the four candidate drugs in cell-culture system has been confirmed.
|
| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Construction of the infectious HEV progenies expressing nanoKAZ and confirmation of the utility of this screening system have been completed, followed by comprehensive screening assay to FDA-approved drug library which revealed four candidate drugs with anti-HEV activity. Cell-culture evaluation to confirm the effectiveness of the four candidate drugs in vitro has been completed as well.
|
| Strategy for Future Research Activity |
Comprehensive screening to small compound library using the infectious HEV progenies expressing nanoKAZ will be started in the fiscal year 2021. The candidates found in this screening will then be subject to cell-culture evaluation to confirm their effectiveness in vitro. Furthermore, inhibitory mechanism of the candidates revealed in the comprehensive drug screening to FDA-approved drug library and small compound library will be analyzed in cell-culture system.
In addition, screening to the small compound library that binds to HEV particles will be performed to find the small compound that can directly inhibit the attachment of the virus protein to host receptor, followed by analysis of its inhibitory mechanism in vitro.
|
