title: High Throughput Measurement of Ca++ Dynamics in Human Stem Cell-Derived Cardiomyocytes by Kinetic Image Cytometery: A Cardiac Risk Assessment Characterization Using a Large Panel of Cardioactive and Inactive Compounds.
abstract: Human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) are emerging as a powerful in vitro model for cardiac safety assessment which may allow for better identification of compounds with poor arrhythmogenic liability profiles early in the drug discovery process. Here, we describe our examination of the Kinetic Image Cytometer (KIC) system's ability to predict adverse compound effects using hiPS-CMs and a library of 53 compounds, the majority of which are known to be cardioactive compounds, and several negative controls. The KIC provides a high throughput method for analyzing intracellular calcium transients. In the cardiomyocyte, intracellular calcium transients integrate the electrochemical signals of the action potential (AP) with the molecular signaling pathways regulating contraction. Drug-induced alterations in the shape and duration of AP result in changes to the shape and duration of the intracellular calcium transient. By examining calcium transient dynamics in hiPS-CMs, KIC can be used as a phenotypic screen to assess compound effects across multiple ion channel types (MITs), detecting MITs, calcium handling and signaling effects. The results of this blinded study indicate that using hiPS-CMs, KIC is able to accurately detect drug-induced changes in Ca(2+) transient dynamics (ie, duration and beat rate) and therefore, may be useful in predicting drug-induced arrhythmogenic liabilities in early de-risking within the drug discovery phase.
authors.affiliation: *Global Safety Pharmacology, Preclinical Development & Safety, Discovery Sciences, Janssen Pharmaceutical NV, B2340 Beerse, Belgium; hlu@its.jnj.com.
authors.forename: Hua Rong
authors.initials: HR
authors.lastname: Lu
authors.affiliation: Vala Sciences Inc. San Diego, California 92121;
authors.forename: Ross
authors.initials: R
authors.lastname: Whittaker
authors.affiliation: Vala Sciences Inc. San Diego, California 92121;
authors.forename: Jeffrey H
authors.initials: JH
authors.lastname: Price
authors.affiliation: Vala Sciences Inc. San Diego, California 92121;
authors.forename: Raquel
authors.initials: R
authors.lastname: Vega
authors.affiliation: Vala Sciences Inc. San Diego, California 92121;
authors.forename: Emily R
authors.initials: ER
authors.lastname: Pfeiffer
authors.affiliation: Vala Sciences Inc. San Diego, California 92121;
authors.forename: Fabio
authors.initials: F
authors.lastname: Cerignoli
authors.affiliation: Consultant at Audacter Consulting, Lydney, Gloucestershire, United Kingdom.
authors.forename: Rob
authors.initials: R
authors.lastname: Towart
authors.affiliation: *Global Safety Pharmacology, Preclinical Development & Safety, Discovery Sciences, Janssen Pharmaceutical NV, B2340 Beerse, Belgium;
authors.forename: David J
authors.initials: DJ
authors.lastname: Gallacher
mesh_terms: D000200:Action Potentials; D000826:Animal Testing Alternatives; D020013:Calcium Signaling / Q000187:drug effects*; D066126:Cardiotoxicity; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D006331:Heart Diseases / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D006339:Heart Rate / Q000187:drug effects; D057166:High-Throughput Screening Assays; D006801:Humans; D019044:Image Cytometry; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007700:Kinetics; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010641:Phenotype; D015203:Reproducibility of Results; D018570:Risk Assessment; D018675:Toxicity Tests / Q000379:methods*
keywords: Action potential (AP); Alternatives to animal testing; Calcium transient dynamics; Cardiac arrhythmias; High content screen (HCS); Human ether-à-go-go related gene (HERG); Long QT