TY - JOUR
T1 - Electrophysiological characterization of drug response in hSC-derived cardiomyocytes using voltage-sensitive optical platforms
AU - Pfeiffer-Kaushik, Emily R.
AU - Smith, Godfrey L.
AU - Cai, Beibei
AU - Dempsey, Graham T.
AU - Hortigon-Vinagre, Maria P.
AU - Zamora, Victor
AU - Feng, Shuyun
AU - Ingermanson, Randall
AU - Zhu, Renjun
AU - Hariharan, Venkatesh
AU - Nguyen, Cuong
AU - Pierson, Jennifer
AU - Gintant, Gary A.
AU - Tung, Leslie
N1 - Funding Information:
The Authors would like to acknowledge the HESI Cardiac Safety Committee Myocyte Subteam for their intellectual contributions to this study. Compounds were prepared and distributed by Liang Guo, Alexander Martinkosky and Doug Smallwood (Chemotherapeutic Agents Repository, National Cancer Institute, U.S. Health and Human Services). M.P.H.-V. and V.Z. were recipients of a postdoctoral fellowship from Fundacion Alfonso Martin Escudero, Spain. V.H. was a recipient of NIH postdoctoral fellowship F32 HL128079.
Funding Information:
HESI's scientific initiatives are primarily supported by the in-kind contributions (from public and private sector participants) of time, expertise, and experimental effort, including the cells provided by Cellular Dynamics International and Axiogenesis (now Ncardia). The authors thank Ross Whittaker, Patrick McDonough and Jeffrey Price for discussion and support (Vala Sciences). These contributions are supplemented by direct funding (that primarily support program infrastructure and management) provided by HESI's corporate sponsors. This project was principally supported through in-kind contributions from the participating organizations. In addition, this research was supported in part by the Developmental Therapeutics Program in the Division of Cancer Treatment and Diagnosis of the National Cancer Institute. Also, this project was funded in part with federal funds from the National Cancer Institute, National Institutes of Health, under contract no. HHSN261200800001E and R24 HL117756.
Funding Information:
The Authors would like to acknowledge the HESI Cardiac Safety Committee Myocyte Subteam for their intellectual contributions to this study. Compounds were prepared and distributed by Liang Guo, Alexander Martinkosky and Doug Smallwood (Chemotherapeutic Agents Repository, National Cancer Institute, U.S. Health and Human Services). M.P.H.-V. and V.Z. were recipients of a postdoctoral fellowship from Fundacion Alfonso Martin Escudero, Spain. V.H. was a recipient of NIH postdoctoral fellowship F32 HL128079. HESI's scientific initiatives are primarily supported by the in-kind contributions (from public and private sector participants) of time, expertise, and experimental effort, including the cells provided by Cellular Dynamics International and Axiogenesis (now Ncardia). The authors thank Ross Whittaker, Patrick McDonough and Jeffrey Price for discussion and support (Vala Sciences). These contributions are supplemented by direct funding (that primarily support program infrastructure and management) provided by HESI's corporate sponsors. This project was principally supported through in-kind contributions from the participating organizations. In addition, this research was supported in part by the Developmental Therapeutics Program in the Division of Cancer Treatment and Diagnosis of the National Cancer Institute. Also, this project was funded in part with federal funds from the National Cancer Institute, National Institutes of Health, under contract no. HHSN261200800001E and R24 HL117756.
Publisher Copyright:
© 2019
PY - 2019
Y1 - 2019
N2 - 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 IKr, ICaL, INa, and IKs, 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.
AB - 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 IKr, ICaL, INa, and IKs, 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.
KW - Action potential
KW - Cardiac electrophysiology
KW - Comprehensive in vitro proarrhythmia assay (CiPA)
KW - ICH S7B
KW - Methods
KW - Safety pharmacology
KW - Stem cell-derived cardiomyocyte
KW - Torsades de pointes (TdP) arrhythmia
KW - Voltage-sensitive optical sensors
KW - hERG
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U2 - 10.1016/j.vascn.2019.106612
DO - 10.1016/j.vascn.2019.106612
M3 - Article
C2 - 31319140
AN - SCOPUS:85068253518
SN - 1056-8719
VL - 99
JO - Journal of Pharmacological and Toxicological Methods
JF - Journal of Pharmacological and Toxicological Methods
M1 - 106612
ER -