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dc.contributor.advisorAdamson, Catherine S.
dc.contributor.advisorRandall, R. E.
dc.contributor.authorVasou, Andri
dc.coverage.spatialxviii, 233 p.en_US
dc.description.abstractAll viruses encode for at least one viral interferon (IFN) antagonist, which is used to subvert the cellular IFN response, a powerful antiviral innate immune response. Numerous in vitro and in vivo studies have demonstrated that IFN antagonism is crucial for virus survival, suggesting that viral IFN antagonists could represent promising therapeutic targets. This study focuses on Respiratory Syncytial Virus (RSV), an important human pathogen for which there is no vaccine or virus-specific antiviral drug. RSV encodes two IFN antagonists NS1 and NS2, which play a critical role in RSV replication and pathogenicity. We developed a high-throughput screening (HTS) assay to target NS2 via our cell-line, which contains a GFP gene under the control of an IFN-stimulated response element (ISRE) to monitor IFN- signalling pathway. NS2 inhibits the IFN-signalling pathway and hence GFP expression in the cell-line by mediating STAT2 degradation. Using a HTS approach, we screened 16,000 compounds to identify small molecules that inhibit NS2 function and therefore relinquish the NS2 imposed block to IFN-signalling, leading to restoration of GFP expression. A total of twenty-eight hits were identified; elimination of false positives left eight hits, four of which (AV-14, -16, -18, -19) are the most promising. These four hit compounds have EC₅₀ values in the single μM range and three of them (AV-14, -16, -18) represent a chemically related series with an indole structure. We demonstrated that the hit compounds specifically inhibit the STAT2 degradation function of NS2, not the function of NS1 or unrelated viral IFN antagonists. At the current time, compounds do not restrict RSV replication in vitro, hence hit optimization is required to improve their potency. Nonetheless, these compounds could be used as chemical tools to determine the unknown mechanism by which NS2 mediates STAT2 degradation and tackle fundamental questions about RSV biology.en_US
dc.publisherUniversity of St Andrews
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
dc.subjectRespiratory Syncytial Virus (RSV)en_US
dc.subjectRSV antiviralsen_US
dc.subjectViral interferon (IFN) antagonistsen_US
dc.subjectRSV IFN antagonistsen_US
dc.subjectRSV NS1en_US
dc.subjectRSV NS2en_US
dc.subjectRSV NS2 inhibitorsen_US
dc.subjectSTAT2 degradationen_US
dc.subjectIFN-signalling pathwayen_US
dc.subjectCell-based high-throughput screening (HTS) assayen_US
dc.subjectGFP reporter assayen_US
dc.subject.lcshMicrobiological assayen_US
dc.subject.lcshViral proteinsen_US
dc.subject.lcshRespiratory syncytial virusen_US
dc.titleDevelopment of a novel cell-based screening platform to identify inhibitors of viral interferon antagonists from clinically important virusesen_US
dc.type.qualificationnamePhD Doctor of Philosophyen_US
dc.publisher.institutionThe University of St Andrewsen_US

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Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Except where otherwise noted within the work, this item's license for re-use is described as Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International