title: Cell-Based High-Throughput Screening Assay Identifies 2',2'-Difluoro-2'-deoxycytidine Gemcitabine as a Potential Antipoliovirus Agent.
abstract: As we approach the global eradication of circulating wild-type polioviruses (PV), vaccination with oral poliovirus vaccine (OPV) has led to the emergence of circulating vaccine-derived poliovirus (cVDPV) and vaccine-associated paralytic poliomyelitis (VAPP). Complete cessation of all poliovirus infections may require stopping use of OPV and formulating improved vaccines and new antiviral drugs. Currently, no licensed drugs are available to treat chronically infected poliovirus excretors. Here, we created a modified PV expressing Gaussia Luciferase (Sb-Gluc) and developed a cell-based high-throughput screening (HTS) antiviral assay. Using the validated HTS assay, we screened the FDA-approved drug library of compounds and identified candidate agents capable of inhibiting PV replication. We then characterized antipoliovirus activity for the best hit, gemcitabine, a nucleoside analogue used in tumor chemotherapy. We found that gemcitabine inhibited PV Mahoney replication with an IC50of 0.3 μM. It completely protected HeLa cells from PV-induced cytopathic effects at 25 μM, without detectable toxicity for cell viability. Furthermore, a gemcitabine metabolite directly inhibited the ability of PV RNA polymerase to synthesize or elongate PV RNA. Because PV RNA polymerase is somehow conserved among species in the Picornaviridae family, gemcitabine may be further developed as an attractive broad-spectrum antiviral for PV and others.
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Zhuoran
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.|Unit of anti-tuberculosis immunity, CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences , Shanghai 200031, China.
authors.forename: Enzhuo
authors.initials: E
authors.lastname: Yang
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Chunmiao
authors.initials: C
authors.lastname: Hu
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Han
authors.initials: H
authors.lastname: Cheng
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Crystal Y
authors.initials: CY
authors.lastname: Chen
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Dan
authors.initials: D
authors.lastname: Huang
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Richard
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Lijun
authors.initials: L
authors.lastname: Rong
authors.affiliation: Viral Populations and Pathogenesis Unit, CNRS UMR 3569, Institut Pasteur , 25-28 rue du Dr. Roux, 75724 Paris cedex 15, France.
authors.forename: Marco
authors.initials: M
authors.lastname: Vignuzzi
authors.affiliation: Unit of anti-tuberculosis immunity, CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences , Shanghai 200031, China.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Shen
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.
authors.forename: Ling
authors.initials: L
authors.lastname: Shen
authors.affiliation: Department of Microbiology and Immunology and Center for Primate Biomedical Research, University of Illinois College of Medicine , 909 South Wolcott Avenue, E704, M/C790, Chicago, Illinois 60612, United States.|Institut Pasteur of Shanghai, Shanghai 200031, China.
authors.forename: Zheng W
authors.initials: ZW
authors.lastname: Chen
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D017955:Poliovirus / Q000187:drug effects*; D010948:Viral Plaque Assay; D014779:Virus Replication; D000093542:Gemcitabine
chemical_list: D000998:Antiviral Agents; D003841:Deoxycytidine; D000093542:Gemcitabine
keywords: RNA-dependent RNA polymerase inhibitor; antiviral; gemcitabine; poliovirus