Human induced pluripotent stem cell-derived cardiomyocytes as an in vitro model for coxsackievirus B3-induced myocarditis and antiviral drug screening platform

Circ Res. 2014 Aug 29;115(6):556-66. doi: 10.1161/CIRCRESAHA.115.303810. Epub 2014 Jul 11.

Abstract

Rationale: 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.

Methods and 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.

Keywords: myocarditis; myocytes, cardiac; stem cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiviral Agents / pharmacology
  • Antiviral Agents / therapeutic use*
  • Calcium / metabolism
  • Cell Proliferation
  • Cells, Cultured
  • Drug Evaluation, Preclinical
  • Enterovirus B, Human / isolation & purification*
  • Enterovirus Infections / drug therapy*
  • Enterovirus Infections / metabolism
  • Humans
  • In Vitro Techniques
  • Models, Cardiovascular*
  • Myocarditis / drug therapy*
  • Myocarditis / metabolism
  • Myocarditis / virology
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / pathology*
  • Myocytes, Cardiac / virology
  • Pluripotent Stem Cells / drug effects
  • Pluripotent Stem Cells / pathology*
  • Pluripotent Stem Cells / virology
  • RNA, Viral / metabolism
  • Treatment Outcome

Substances

  • Antiviral Agents
  • RNA, Viral
  • Calcium