Abstract Title
High throughput screening on murine norovirus to identify antiviral compounds
Presenter
Johanna Becker, Center for Integrative Infectious Disease Research, Uni Heidelberg, Molecular Virology
Co-Author(s)
J. Becker1, M. Lingemann1, J. Hu2, V. L. Dao Thi2, V. Lohmann1
1University of Heidelberg, Molecular Virology, Heidelberg, Germany
2University of Heidelberg, Virology, Heidelberg, Germany
Abstract Category
Prevention & Control (antivirals)
Abstract
Background
For human noroviruses (HuNoV), the main cause of acute gastroenteritis, no antivirals are available. Since HuNoV is recalcitrant in cell culture, murine norovirus (MNV) is used as a model system. MNV displays similar genomic and protein structures. The expression of the MNV receptor CD300lf renders different cell lines susceptible to infection, increasing its value as a model system for a high-throughput screening (HTS) approach identifying possible antiviral compounds targeting norovirus replication.
Methods
Since MNV undergoes a lytic replication cycle, we developed a HTS approach utilizing the WST-1 cell viability assay. We transduced the human hepatoma cell line Huh7 Lunet to stably express the CD300lf receptor allowing murine norovirus infection. A low MOI of 0.05 and an incubation time of 48 h p.i. was chosen to allow multiple replication cycles of MNV.
Results
The initial screen included a 8.5 k bioactive compound library with inhibitors targeting known host cell proteins. The broad range of inhibitor classes include previously identified norovirus inhibitors serving as a control. This increases the likelihood of compounds inhibiting all norovirus genogroups. We identified ca. 30 hit compounds, 4 of those showed a profound inhibition of MNV replication without detectable cytotoxicity at the chosen 10 μM. One hit, Rupintrivir, has already been described for its inhibitory effect on HuNoV GI.1 and MNV.
Conclusion
Using the established HTS pipeline for MNV, an initial set of potential inhibitors was identified.
Further evaluation and testing for HuNoV inhibition in the present systems, enteroids and GI.1 replicon, will be done.
For human noroviruses (HuNoV), the main cause of acute gastroenteritis, no antivirals are available. Since HuNoV is recalcitrant in cell culture, murine norovirus (MNV) is used as a model system. MNV displays similar genomic and protein structures. The expression of the MNV receptor CD300lf renders different cell lines susceptible to infection, increasing its value as a model system for a high-throughput screening (HTS) approach identifying possible antiviral compounds targeting norovirus replication.
Methods
Since MNV undergoes a lytic replication cycle, we developed a HTS approach utilizing the WST-1 cell viability assay. We transduced the human hepatoma cell line Huh7 Lunet to stably express the CD300lf receptor allowing murine norovirus infection. A low MOI of 0.05 and an incubation time of 48 h p.i. was chosen to allow multiple replication cycles of MNV.
Results
The initial screen included a 8.5 k bioactive compound library with inhibitors targeting known host cell proteins. The broad range of inhibitor classes include previously identified norovirus inhibitors serving as a control. This increases the likelihood of compounds inhibiting all norovirus genogroups. We identified ca. 30 hit compounds, 4 of those showed a profound inhibition of MNV replication without detectable cytotoxicity at the chosen 10 μM. One hit, Rupintrivir, has already been described for its inhibitory effect on HuNoV GI.1 and MNV.
Conclusion
Using the established HTS pipeline for MNV, an initial set of potential inhibitors was identified.
Further evaluation and testing for HuNoV inhibition in the present systems, enteroids and GI.1 replicon, will be done.