title: A phenotypic assay to identify Chikungunya virus inhibitors targeting the nonstructural protein nsP2.
abstract: Chikungunya virus (CHIKV) is a mosquito-transmitted pathogen responsible for an acute infection of abrupt onset, characterized by high fever, polyarthralgia, myalgia, headaches, chills, and rash. In 2006, CHIKV was responsible for an epidemic outbreak of unprecedented magnitude in the Indian Ocean, stressing the need for therapeutic approaches. Since then, we have acquired a better understanding of CHIKV biology, but we are still missing active molecules against this reemerging pathogen. We recently reported that the nonstructural nsP2 protein of CHIKV induces a transcriptional shutoff that allows the virus to block cellular antiviral response. This was demonstrated using various luciferase-based reporter gene assays, including a trans-reporter system where Gal4 DNA binding domain is fused to Fos transcription factor. Here, we turned this assay into a high-throughput screening system to identify small molecules targeting nsP2-mediated shutoff. Among 3040 molecules tested, we identified one natural compound that partially blocks nsP2 activity and inhibits CHIKV replication in vitro. This proof of concept suggests that similar functional assays could be developed to target other viral proteins mediating a cellular shutoff and identify innovative therapeutic molecules.
authors.affiliation: Institut Pasteur, Unité de Génomique Virale et Vaccination, Paris, France.
authors.forename: Marianne
authors.initials: M
authors.lastname: Lucas-Hourani
authors.forename: Alexandru
authors.initials: A
authors.lastname: Lupan
authors.forename: Philippe
authors.initials: P
authors.lastname: Desprès
authors.forename: Sylviane
authors.initials: S
authors.lastname: Thoret
authors.forename: Olivier
authors.initials: O
authors.lastname: Pamlard
authors.forename: Joëlle
authors.initials: J
authors.lastname: Dubois
authors.forename: Catherine
authors.initials: C
authors.lastname: Guillou
authors.forename: Frédéric
authors.initials: F
authors.lastname: Tangy
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Vidalain
authors.forename: Hélène
authors.initials: H
authors.lastname: Munier-Lehmann
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D002646:Chikungunya virus / Q000187:drug effects* / Q000502:physiology*; D004353:Drug Evaluation, Preclinical; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D010641:Phenotype; D015203:Reproducibility of Results; D054852:Small Molecule Libraries; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D054852:Small Molecule Libraries; D017361:Viral Nonstructural Proteins