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

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.

• Authors: Arun Sharma, Caleb Marceau, Ryoko Hamaguchi, Paul W Burridge, Kuppusamy Rajarajan, Jared M Churko, Haodi Wu, Karim I Sallam, Elena Matsa, Anthony C Sturzu, Yonglu Che, Antje Ebert, Sebastian Diecke, Ping Liang, Kristy Red-Horse, Jan E Carette, Sean M Wu, Joseph C Wu
• Journal: Circulation research (Circ Res) Vol. 115 Issue 6 Pg. 556-66 (Aug 29 2014) ISSN: 1524-4571 [Electronic] United States
• PMID: 25015077 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 American Heart Association, Inc.

Chemical References

• Antiviral Agents
• RNA, Viral
• Calcium

Topics

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