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dc.contributor.authorVasou, Andri
dc.contributor.authorPaulus, Christina
dc.contributor.authorNarloch, Janina
dc.contributor.authorGage, Zoe O.
dc.contributor.authorRameix-Welti, Marie-Anne
dc.contributor.authorEléouët, Jean-François
dc.contributor.authorNevels, Michael
dc.contributor.authorRandall, Richard E.
dc.contributor.authorAdamson, Catherine S.
dc.identifier.citationVasou , A , Paulus , C , Narloch , J , Gage , Z O , Rameix-Welti , M-A , Eléouët , J-F , Nevels , M , Randall , R E & Adamson , C S 2018 , ' Modular cell-based platform for high throughput identification of compounds that inhibit a viral interferon antagonist of choice ' , Antiviral Research , vol. 150 , pp. 79-92 .
dc.identifier.otherORCID: /0000-0003-4051-4658/work/54516601
dc.identifier.otherORCID: /0000-0002-7115-407X/work/37898080
dc.identifier.otherORCID: /0000-0002-4123-5629/work/47136633
dc.identifier.otherORCID: /0000-0002-9304-6678/work/60426968
dc.identifier.otherORCID: /0000-0001-7673-5212/work/60630448
dc.descriptionThe work was supported by the Medical Research Council, U.K. (University of St Andrews Doctoral Training Grant to AV and CSA), Deutsche Forschungsgemeinschaft (PA 815/2-1) to CP, Tenovus Scotland (T15/38) to MN and Wellcome Trust to CP, MN (ISSF) and RER (101788/Z/13/Z)en
dc.description.abstractViral interferon (IFN) antagonists are a diverse class of viral proteins that counteract the host IFN response, which is important for controlling viral infections. Viral IFN antagonists are often multifunctional proteins that perform vital roles in virus replication beyond IFN antagonism. The critical importance of viral IFN antagonists is highlighted by the fact that almost all viruses encode one of these proteins. Inhibition of viral IFN antagonists has the potential to exert pleiotropic antiviral effects and thus this important protein class represents a diverse plethora of novel therapeutic targets. To exploit this, we have successfully developed and executed a novel modular cell-based platform that facilitates the safe and rapid screening for inhibitors of a viral IFN antagonist of choice. The platform is based on two reporter cell-lines that provide a simple method to detect activation of IFN induction or signaling via an eGFP gene placed under the control of the IFNβ or an ISRE-containing promoter, respectively. Expression of a target IFN antagonist in the appropriate reporter cell-line will block the IFN response and hence eGFP expression. We hypothesized that addition of a compound that inhibits IFN antagonist function will release the block imposed on the IFN response and hence restore eGFP expression, providing a measurable parameter for high throughput screening (HTS). We demonstrate assay proof-of-concept by (i) exploiting hepatitis C virus (HCV) protease inhibitors to inhibit NS3-4A's capacity to block IFN induction and (ii) successfully executing two HTS targeting viral IFN antagonists that block IFN signaling; NS2 and IE1 from human respiratory syncytial virus (RSV) and cytomegalovirus (CMV) respectively, two clinically important viruses for which vaccine development has thus far been unsuccessful and new antivirals are required. Both screens performed robustly and Z′ Factor scores of >0.6 were achieved. We identified (i) four hit compounds that specifically inhibit RSV NS2's ability to block IFN signaling by mediating STAT2 degradation and exhibit modest antiviral activity and (ii) two hit compounds that interfere with IE1 transcription and significantly impair CMV replication. Overall, we demonstrate assay proof-of-concept as we target viral IFN antagonists from unrelated viruses and demonstrate its suitability for HTS.
dc.relation.ispartofAntiviral Researchen
dc.subjectViral interferon (IFN) antagonistsen
dc.subjectHuman Respiratory Syncytial Virus (RSV)en
dc.subjectHuman Cytomegalovirus (CMV)en
dc.subjectHigh-throughput screening (HTS)en
dc.subjectSignal transducer and activator of transcription 2 (STAT2)en
dc.subjectQR355 Virologyen
dc.subjectQR180 Immunologyen
dc.subjectQH426 Geneticsen
dc.subjectBiochemistry, Genetics and Molecular Biology(all)en
dc.subjectImmunology and Microbiology(all)en
dc.subjectInfectious Diseasesen
dc.subjectSDG 3 - Good Health and Well-beingen
dc.titleModular cell-based platform for high throughput identification of compounds that inhibit a viral interferon antagonist of choiceen
dc.typeJournal articleen
dc.contributor.sponsorThe Wellcome Trusten
dc.contributor.institutionUniversity of St Andrews. School of Biologyen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.description.statusPeer revieweden

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