title: Cross-resistance mechanism of respiratory syncytial virus against structurally diverse entry inhibitors.
abstract: Respiratory syncytial virus (RSV) is a leading pediatric pathogen that is responsible for a majority of infant hospitalizations due to viral disease. Despite its clinical importance, no vaccine prophylaxis against RSV disease or effective antiviral therapeutic is available. In this study, we established a robust high-throughput drug screening protocol by using a recombinant RSV reporter virus to expand the pool of RSV inhibitor candidates. Mechanistic characterization revealed that a potent newly identified inhibitor class blocks viral entry through specific targeting of the RSV fusion (F) protein. Resistance against this class was induced and revealed overlapping hotspots with diverse, previously identified RSV entry blockers at different stages of preclinical and clinical development. A structural and biochemical assessment of the mechanism of unique, broad RSV cross-resistance against structurally distinct entry inhibitors demonstrated that individual escape hotspots are located in immediate physical proximity in the metastable conformation of RSV F and that the resistance mutations lower the barrier for prefusion F triggering, resulting in an accelerated RSV entry kinetics. One resistant RSV recombinant remained fully pathogenic in a mouse model of RSV infection. By identifying molecular determinants governing the RSV entry machinery, this study spotlights a molecular mechanism of broad RSV resistance against entry inhibition that may affect the impact of diverse viral entry inhibitors presently considered for clinical use and outlines a proactive design for future RSV drug discovery campaigns.
authors.affiliation: Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303;
authors.forename: Dan
authors.initials: D
authors.lastname: Yan
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322;Children's Healthcare of Atlanta, Atlanta, GA 30322; and.
authors.forename: Sujin
authors.initials: S
authors.lastname: Lee
authors.affiliation: Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303;
authors.forename: Vidhi D
authors.initials: VD
authors.lastname: Thakkar
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL 35294.
authors.forename: Ming
authors.initials: M
authors.lastname: Luo
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322;Children's Healthcare of Atlanta, Atlanta, GA 30322; and.
authors.forename: Martin L
authors.initials: ML
authors.lastname: Moore
authors.affiliation: Institute for Biomedical Sciences, Georgia State University, Atlanta, GA 30303;Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322; rplemper@gsu.edu.
authors.forename: Richard Karl
authors.initials: RK
authors.lastname: Plemper
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D006224:Cricetinae; D024882:Drug Resistance, Viral; D008561:Membrane Fusion / Q000502:physiology; D008807:Mice, Inbred BALB C; D009154:Mutation; D012136:Respiratory Syncytial Viruses / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D054852:Small Molecule Libraries; D014764:Viral Proteins / Q000235:genetics
chemical_list: D000998:Antiviral Agents; D054852:Small Molecule Libraries; D014764:Viral Proteins
keywords: antiviral drugs; drug resistance
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