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

A Sharma, C Marceau, R Hamaguchi… - Circulation …, 2014 - Am Heart Assoc
A Sharma, C Marceau, R Hamaguchi, PW Burridge, K Rajarajan, JM Churko, H Wu
Circulation research, 2014Am Heart Assoc
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 …
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.
Am Heart Assoc