TY - JOUR
T1 - An interferon-beta promoter reporter assay for high throughput identification of compounds against multiple RNA viruses
AU - Guo, Fang
AU - Zhao, Xuesen
AU - Gill, Tina
AU - Zhou, Yan
AU - Campagna, Matthew
AU - Wang, Lijuan
AU - Liu, Fei
AU - Zhang, Pinghu
AU - Dipaolo, Laura
AU - Du, Yanming
AU - Xu, Xiaodong
AU - Jiang, Dong
AU - Wei, Lai
AU - Cuconati, Andrea
AU - Block, Timothy M.
AU - Guo, Ju Tao
AU - Chang, Jinhong
N1 - Copyright © 2014 Elsevier B.V. All rights reserved.
PY - 2014/7
Y1 - 2014/7
N2 - 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.
AB - 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.
KW - Antiviral
KW - Dengue virus
KW - High throughput assay
KW - Innate immune modulator
KW - Artificial Gene Fusion
KW - Cell Line
KW - Promoter Regions, Genetic
KW - Antiviral Agents/isolation & purification
KW - Humans
KW - Interferon-beta/biosynthesis
KW - Luciferases, Firefly/analysis
KW - High-Throughput Screening Assays
KW - Drug Evaluation, Preclinical/methods
KW - RNA Viruses/drug effects
KW - Genes, Reporter
KW - Immunologic Factors/isolation & purification
UR - http://www.scopus.com/inward/record.url?scp=84901058059&partnerID=8YFLogxK
UR - https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=purepublist2023&SrcAuth=WosAPI&KeyUT=WOS:000338412500009&DestLinkType=FullRecord&DestApp=WOS
U2 - 10.1016/j.antiviral.2014.04.010
DO - 10.1016/j.antiviral.2014.04.010
M3 - Article
C2 - 24792753
SN - 0166-3542
VL - 107
SP - 56
EP - 65
JO - Antiviral Research
JF - Antiviral Research
IS - 1
ER -