title: Human induced pluripotent stem cell-derived cardiomyocytes as an in vitro model for coxsackievirus B3-induced myocarditis and antiviral drug screening platform.
abstract: BACKGROUND
  Viral myocarditis is a life-threatening illness that may lead to heart failure or cardiac arrhythmias. A major causative agent for viral myocarditis is the B3 strain of coxsackievirus, a positive-sense RNA enterovirus. However, human cardiac tissues are difficult to procure in sufficient enough quantities for studying the mechanisms of cardiac-specific viral infection.
  
  
  OBJECTIVE
  This study examined whether human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) could be used to model the pathogenic processes of coxsackievirus-induced viral myocarditis and to screen antiviral therapeutics for efficacy.
  
  
  RESULTS
  hiPSC-CMs were infected with a luciferase-expressing coxsackievirus B3 strain (CVB3-Luc). Brightfield microscopy, immunofluorescence, and calcium imaging were used to characterize virally infected hiPSC-CMs for alterations in cellular morphology and calcium handling. Viral proliferation in hiPSC-CMs was quantified using bioluminescence imaging. Antiviral compounds including interferonβ1, ribavirin, pyrrolidine dithiocarbamate, and fluoxetine were tested for their capacity to abrogate CVB3-Luc proliferation in hiPSC-CMs in vitro. The ability of these compounds to reduce CVB3-Luc proliferation in hiPSC-CMs was consistent with reported drug effects in previous studies. Mechanistic analyses via gene expression profiling of hiPSC-CMs infected with CVB3-Luc revealed an activation of viral RNA and protein clearance pathways after interferonβ1 treatment.
  
  
  CONCLUSIONS
  This study demonstrates that hiPSC-CMs express the coxsackievirus and adenovirus receptor, are susceptible to coxsackievirus infection, and can be used to predict antiviral drug efficacy. Our results suggest that the hiPSC-CM/CVB3-Luc assay is a sensitive platform that can screen novel antiviral therapeutics for their effectiveness in a high-throughput fashion.
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Arun
authors.initials: A
authors.lastname: Sharma
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Caleb
authors.initials: C
authors.lastname: Marceau
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Hamaguchi
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Paul W
authors.initials: PW
authors.lastname: Burridge
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Kuppusamy
authors.initials: K
authors.lastname: Rajarajan
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Jared M
authors.initials: JM
authors.lastname: Churko
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Haodi
authors.initials: H
authors.lastname: Wu
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Karim I
authors.initials: KI
authors.lastname: Sallam
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Elena
authors.initials: E
authors.lastname: Matsa
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Anthony C
authors.initials: AC
authors.lastname: Sturzu
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Yonglu
authors.initials: Y
authors.lastname: Che
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Antje
authors.initials: A
authors.lastname: Ebert
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Diecke
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Ping
authors.initials: P
authors.lastname: Liang
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Kristy
authors.initials: K
authors.lastname: Red-Horse
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA.
authors.forename: Jan E
authors.initials: JE
authors.lastname: Carette
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA. joewu@stanford.edu smwu@stanford.edu.
authors.forename: Sean M
authors.initials: SM
authors.lastname: Wu
authors.affiliation: From the Department of Medicine, Division of Cardiology (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Institute for Stem Cell Biology and Regenerative Medicine (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., S.M.W., J.C.W.), Stanford Cardiovascular Institute (A.S., R.H., P.W.B., K.R., J.M.C., H.W., K.I.S., E.M., A.C.S., Y.C., A.E., S.D., P.L., K.R.-H., S.M.W., J.C.W.), Department of Biology (A.S., R.H., K.R.-H.), Department of Microbiology and Immunology (C.M., J.E.C.), and Department of Radiology, Molecular Imaging Program (J.C.W.), Stanford University School of Medicine, CA. joewu@stanford.edu smwu@stanford.edu.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Wu
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology / Q000627:therapeutic use*; D002118:Calcium / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D029822:Enterovirus B, Human / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000188:drug therapy* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008955:Models, Cardiovascular; D009205:Myocarditis / Q000188:drug therapy* / Q000378:metabolism / Q000821:virology; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000473:pathology* / Q000821:virology; D039904:Pluripotent Stem Cells / Q000187:drug effects / Q000473:pathology* / Q000821:virology; D012367:RNA, Viral / Q000378:metabolism; D016896:Treatment Outcome
chemical_list: D000998:Antiviral Agents; D012367:RNA, Viral; D002118:Calcium
keywords: myocarditis; myocytes, cardiac; stem cells