title: An interferon-beta promoter reporter assay for high throughput identification of compounds against multiple RNA viruses.
abstract: Virus infection of host cells is sensed by innate pattern recognition receptors (PRRs) and induces production of type I interferons (IFNs) and other inflammatory cytokines. These cytokines orchestrate the elimination of the viruses but are occasionally detrimental to the hosts. The outcomes and pathogenesis of viral infection are largely determined by the specific interaction between the viruses and their host cells. Therefore, compounds that either inhibit viral infection or modulate virus-induced cytokine response should be considered as candidates for managing virus infection. The aim of the study was to identify compounds in both categories, using a single cell-based assay. Our screening platform is a HEK293 cell-based reporter assay where the expression of a firefly luciferase is under the control of a human IFN-β promoter. We have demonstrated that infection of the reporter cell line with a panel of RNA viruses activated the reporter gene expression that correlates quantitatively with the levels of virus replication and progeny virus production, and could be inhibited in a dose-dependent manner by known antiviral compound or inhibitors of PRR signal transduction pathways. Using Dengue virus as an example, a pilot screening of a small molecule library consisting of 26,900 compounds proved the concept that the IFN-β promoter reporter assay can serve as a convenient high throughput screening platform for simultaneous discovery of antiviral and innate immune response modulating compounds. A representative antiviral compound from the pilot screening, 1-(6-ethoxybenzo[d]thiazol-2-yl)-3-(3-methoxyphenyl) urea, was demonstrated to specifically inhibit several viruses belonging to the family of flaviviridae.
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Guo
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Xuesen
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Tina
authors.initials: T
authors.lastname: Gill
authors.affiliation: Biostatistics and Bioinformatics Facility, Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Campagna
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Wang
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Liu
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Pinghu
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: DiPaolo
authors.affiliation: Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, PA, USA.
authors.forename: Yanming
authors.initials: Y
authors.lastname: Du
authors.affiliation: Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, PA, USA.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Xu
authors.affiliation: Hepatology Institute, Peking University, Beijing, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Jiang
authors.affiliation: Hepatology Institute, Peking University, Beijing, China.
authors.forename: Lai
authors.initials: L
authors.lastname: Wei
authors.affiliation: Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, PA, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Cuconati
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA; Baruch S. Blumberg Institute, Hepatitis B Foundation, Doylestown, PA, USA.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Block
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA.
authors.forename: Ju-Tao
authors.initials: JT
authors.lastname: Guo
authors.affiliation: Institute for Biotechnology and Virology Research, Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, PA, USA. Electronic address: jinhong.chang@drexelmed.edu.
authors.forename: Jinhong
authors.initials: J
authors.lastname: Chang
mesh_terms: D000998:Antiviral Agents / Q000302:isolation & purification*; D019677:Artificial Gene Fusion; D002460:Cell Line; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays; D006801:Humans; D007155:Immunologic Factors / Q000302:isolation & purification*; D016899:Interferon-beta / Q000096:biosynthesis* / Q000235:genetics; D049409:Luciferases, Firefly / Q000032:analysis / Q000235:genetics; D011401:Promoter Regions, Genetic; D012328:RNA Viruses / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D007155:Immunologic Factors; D016899:Interferon-beta; D049409:Luciferases, Firefly
keywords: Antiviral; Dengue virus; High throughput assay; Innate immune modulator