Electrophysiological characterization of drug response in hSC-derived cardiomyocytes using voltage-sensitive optical platforms

Emily R Pfeiffer-Kaushik; Godfrey L Smith; Beibei Cai; Graham T Dempsey; Maria P Hortigon-Vinagre; Victor Zamora; Shuyun Feng; Randall Ingermanson; Renjun Zhu; Venkatesh Hariharan; Cuong Nguyen; Jennifer Pierson; Gary A Gintant; Leslie Tung

doi:10.1016/j.vascn.2019.106612

Electrophysiological characterization of drug response in hSC-derived cardiomyocytes using voltage-sensitive optical platforms

J Pharmacol Toxicol Methods. 2019 Sep-Oct:99:106612. doi: 10.1016/j.vascn.2019.106612. Epub 2019 Jul 15.

Affiliations

¹ Vala Sciences Inc., 6370 Nancy Ridge Drive, Suite 106, San Diego, CA 92121, USA.
² Clyde Biosciences Ltd, BioCity Scotland, Bo'Ness Road, Newhouse, Lanarkshire, Scotland ML1 5UH, United Kingdom; Institute of Cardiovascular and Medical Sciences, College of Medical, Veterinary and Life Science, University of Glasgow, 126 University Place, Glasgow G12 8TA, United Kingdom.
³ Q-State Biosciences Inc., 179 Sidney Street, Cambridge, MA 02139, USA.
⁴ Department of Biomedical Engineering, The Johns Hopkins University, 720 Rutland Ave., Baltimore, MD 21205, USA.
⁵ Health and Environmental Sciences Institute, Washington, D.C. 20009, USA. Electronic address: jpierson@hesiglobal.org.
⁶ AbbVie, 1 North Waukegan Road, Department ZR-13, Building AP-9A, North Chicago, IL 60064-6119, USA.

PMID: 31319140
DOI: 10.1016/j.vascn.2019.106612

Abstract

Introduction: Voltage-sensitive optical (VSO) sensors offer a minimally invasive method to study the time course of repolarization of the cardiac action potential (AP). This Comprehensive in vitro Proarrhythmia Assay (CiPA) cross-platform study investigates protocol design and measurement variability of VSO sensors for preclinical cardiac electrophysiology assays.

Methods: Three commercial and one academic laboratory completed a limited study of the effects of 8 blinded compounds on the electrophysiology of 2 commercial lines of human induced pluripotent stem-cell derived cardiomyocytes (hSC-CMs). Acquisition technologies included CMOS camera and photometry; fluorescent voltage sensors included di-4-ANEPPS, FluoVolt and genetically encoded QuasAr2. The experimental protocol was standardized with respect to cell lines, plating and maintenance media, blinded compounds, and action potential parameters measured. Serum-free media was used to study the action of drugs, but the exact composition and the protocols for cell preparation and drug additions varied among sites.

Results: Baseline AP waveforms differed across platforms and between cell types. Despite these differences, the relative responses to four selective ion channel blockers (E-4031, nifedipine, mexiletine, and JNJ 303 blocking I_Kr, I_CaL, I_Na, and I_Ks, respectively) were similar across all platforms and cell lines although the absolute changes differed. Similarly, four mixed ion channel blockers (flecainide, moxifloxacin, quinidine, and ranolazine) had comparable effects in all platforms. Differences in repolarisation time course and response to drugs could be attributed to cell type and experimental method differences such as composition of the assay media, stimulated versus spontaneous activity, and single versus cumulative compound addition.

Discussion: In conclusion, VSOs represent a powerful and appropriate method to assess the electrophysiological effects of drugs on iPSC-CMs for the evaluation of proarrhythmic risk. Protocol considerations and recommendations are provided toward standardizing conditions to reduce variability of baseline AP waveform characteristics and drug responses.

Keywords: Action potential; Cardiac electrophysiology; Comprehensive in vitro proarrhythmia assay (CiPA); ICH S7B; Methods; Safety pharmacology; Stem cell-derived cardiomyocyte; Torsades de pointes (TdP) arrhythmia; Voltage-sensitive optical sensors; hERG.