Electrophysiological Characterization of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes for the Development of New In Vitro Assays in Preclinical Cardiac Risk Assessment of Drugs in the context of CiPA
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1
Natural and Medical Sciences Institute, Cell Biology, Germany
Unexpected cardiotoxicity has been the most common cause of market drug withdrawals or developmental termination of promising drug candidates in recent years. An early detection of drug-induced cardiac side effects is desired as it prevents cost-intensive late-stage withdrawals and ensures the patients’ safety. However, current assay systems lack the ability of accurately mimicking human physiological conditions for a sufficient and reliable predictivity. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) together with the microelectrode array (MEA) are proposed to be an efficient and more sensitive model for the investigation of drug-induced cardiotoxicity. However, current guidelines of the US Food and Drug Administration (FDA) for the approval of new pharmaceutical substances are very strict and one-sided, which could lead to the unnecessary withdrawal of promising drugs or in turn, the accidentally classification of a harmful drug as safe. Therefore, the Comprehensive in vitro Proarrhythmia Study (CiPA), initiated by the ILSI Health and Environmental Sciences Institute, aims for the improvement of such guidelines by innovative assays.
As member of the CiPA Myocyte Core Team we tested 28 blinded compounds on a hiPSC-CMs MEA system to evaluate the reliability of the assay. The data was obtained in parallel at several labs and assessed in terms of predictivity and inter-lab reliability. Experiments were performed using two commercially available hiPSC-CM cell lines iCell CM2 from Cellular Dynamics CDI (iCells2) and Cor.4U CM from Axiogenesis (Cor.4U). The electrophysiological phenotype and performance as well as predictivity of compound effects and arrhythmia formation were determined. Both cell lines exhibited a stable and reproducible, but distinct electrophysiological profile. Both cell lines showed similar results in the pharmacologically induced changes in the field potential duration (FPD). However, iCells2 showed better predictivity in arrhythmia formation and risk categorization in our hands compared to Cor.4U.
Furthermore the influence of the cell seeding density on electrophysiological phenotype and compound sensitivity was investigated. iCells2 were seeded at different densities and additional compound tests were performed using E-4031 as test compound. Significant differences in the development of electrophysiological properties such as FPD and Na+ amplitudes could be observed between individual seeding densities. Furthermore, cells at lower densities exhibited enhanced sensitivity to E-4031, whereas in very high densities, almost no effect could be detected. Moreover, remarkable variabilities in the occurrence of arrhythmic events were detected, indicating that the correct seeding density is a critical factor for the outcome of compound tests and large-scale studies like CiPA.
Keywords:
hiPSC-derived cardiomyocytes,
CIPA,
microelectrode array,
compound test,
Assay development,
Proarrhythmia,
safety pharmacology
Conference:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays, Reutlingen, Germany, 4 Jul - 6 Jul, 2018.
Presentation Type:
Oral Presentation
Topic:
Assay development
Citation:
Kuechler
S and
Kraushaar
U
(2019). Electrophysiological Characterization of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes for the Development of New In Vitro Assays in Preclinical Cardiac Risk Assessment of Drugs in the context of CiPA.
Conference Abstract:
MEA Meeting 2018 | 11th International Meeting on Substrate Integrated Microelectrode Arrays.
doi: 10.3389/conf.fncel.2018.38.00019
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Received:
14 May 2018;
Published Online:
17 Jan 2019.
*
Correspondence:
Dr. Udo Kraushaar, Natural and Medical Sciences Institute, Cell Biology, Reutlingen, 72770, Germany, udo.kraushaar@nmi.de