Multicenter Study

. 2018 Sep 25;24(13):3582-3592.

doi: 10.1016/j.celrep.2018.08.079.

International Multisite Study of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes for Drug Proarrhythmic Potential Assessment

Ksenia Blinova¹, Qianyu Dang², Daniel Millard³, Godfrey Smith⁴, Jennifer Pierson⁵, Liang Guo⁶, Mathew Brock⁷, Hua Rong Lu⁸, Udo Kraushaar⁹, Haoyu Zeng¹⁰, Hong Shi¹¹, Xiaoyu Zhang¹², Kohei Sawada¹³, Tomoharu Osada¹⁴, Yasunari Kanda¹⁵, Yuko Sekino¹⁶, Li Pang¹⁷, Tromondae K Feaster¹⁸, Ralf Kettenhofen¹⁹, Norman Stockbridge²⁰, David G Strauss²¹, Gary Gintant²²

Affiliations

¹ Division of Biomedical Physics, Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, US Food and Drug Administration, Silver Spring, MD 20993, USA. Electronic address: ksenia.blinova@fda.hhs.gov.
² Office of Biostatistics, Office of Translational Science, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USA.
³ Axion BioSystems, Atlanta, GA 30309, USA.
⁴ University of Glasgow, Glasgow G12 8QQ, Scotland, UK; Clyde Biosciences, Newhouse ML1 5UH, Scotland, UK.
⁵ Health and Environmental Sciences Institute, Washington, DC 20005, USA.
⁶ Investigative Toxicology, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD 21702, USA.
⁷ Genentech, San Francisco, CA 94080, USA.
⁸ Discovery Sciences, R&D, Janssen Pharmaceutical (JNJ), Beerse, Belgium.
⁹ NMI Natural and Medical Sciences Institute at the University of Tübingen, Reutlingen, Germany.
¹⁰ Merck, Safety & Exploratory Pharmacology Department, West Point, PA 19486, USA.
¹¹ Bristol-Myers Squibb, New York, NY 10154, USA.
¹² ACEA Biosciences, San Diego, CA 92121, USA.
¹³ Eisai, Tsukuba, Ibaraki 300-2635, Japan; The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
¹⁴ LSI Medience, Chiyoda-ku, Tokyo 101-8517, Japan.
¹⁵ Division of Pharmacology, National Institute of Health Sciences, Kawasaki, Kanagawa 210-9501, Japan.
¹⁶ The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; Division of Pharmacology, National Institute of Health Sciences, Kawasaki, Kanagawa 210-9501, Japan.
¹⁷ Division of Biochemical Toxicology, National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA.
¹⁸ Cellular Dynamics International-A FUJIFILM Company, Madison, WI 53711, USA.
¹⁹ Ncardia, Cologne 50829, Germany.
²⁰ Division of Cardiovascular and Renal Products, Office of Drug Evaluation I, Office of New Drugs, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USA.
²¹ Division of Applied and Regulatory Science, Office of Clinical Pharmacology, Office of Translational Science, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USA.
²² AbbVie, North Chicago, IL 60064-6118, USA. Electronic address: gary.gintant@abbvie.com.

PMID: 30257217
PMCID: PMC6226030
DOI: 10.1016/j.celrep.2018.08.079

Multicenter Study

International Multisite Study of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes for Drug Proarrhythmic Potential Assessment

Ksenia Blinova et al. Cell Rep. 2018.

. 2018 Sep 25;24(13):3582-3592.

doi: 10.1016/j.celrep.2018.08.079.

Authors

Affiliations

¹ Division of Biomedical Physics, Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, US Food and Drug Administration, Silver Spring, MD 20993, USA. Electronic address: ksenia.blinova@fda.hhs.gov.
² Office of Biostatistics, Office of Translational Science, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USA.
³ Axion BioSystems, Atlanta, GA 30309, USA.
⁴ University of Glasgow, Glasgow G12 8QQ, Scotland, UK; Clyde Biosciences, Newhouse ML1 5UH, Scotland, UK.
⁵ Health and Environmental Sciences Institute, Washington, DC 20005, USA.
⁶ Investigative Toxicology, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Frederick, MD 21702, USA.
⁷ Genentech, San Francisco, CA 94080, USA.
⁸ Discovery Sciences, R&D, Janssen Pharmaceutical (JNJ), Beerse, Belgium.
⁹ NMI Natural and Medical Sciences Institute at the University of Tübingen, Reutlingen, Germany.
¹⁰ Merck, Safety & Exploratory Pharmacology Department, West Point, PA 19486, USA.
¹¹ Bristol-Myers Squibb, New York, NY 10154, USA.
¹² ACEA Biosciences, San Diego, CA 92121, USA.
¹³ Eisai, Tsukuba, Ibaraki 300-2635, Japan; The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
¹⁴ LSI Medience, Chiyoda-ku, Tokyo 101-8517, Japan.
¹⁵ Division of Pharmacology, National Institute of Health Sciences, Kawasaki, Kanagawa 210-9501, Japan.
¹⁶ The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan; Division of Pharmacology, National Institute of Health Sciences, Kawasaki, Kanagawa 210-9501, Japan.
¹⁷ Division of Biochemical Toxicology, National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR 72079, USA.
¹⁸ Cellular Dynamics International-A FUJIFILM Company, Madison, WI 53711, USA.
¹⁹ Ncardia, Cologne 50829, Germany.
²⁰ Division of Cardiovascular and Renal Products, Office of Drug Evaluation I, Office of New Drugs, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USA.
²¹ Division of Applied and Regulatory Science, Office of Clinical Pharmacology, Office of Translational Science, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD 20993, USA.
²² AbbVie, North Chicago, IL 60064-6118, USA. Electronic address: gary.gintant@abbvie.com.

PMID: 30257217
PMCID: PMC6226030
DOI: 10.1016/j.celrep.2018.08.079

Abstract

To assess the utility of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as an in vitro proarrhythmia model, we evaluated the concentration dependence and sources of variability of electrophysiologic responses to 28 drugs linked to low, intermediate, and high torsades de pointes (TdP) risk categories using two commercial cell lines and standardized protocols in a blinded multisite study using multielectrode array or voltage-sensing optical approaches. Logistical and ordinal linear regression models were constructed using drug responses as predictors and TdP risk categories as outcomes. Three of seven predictors (drug-induced arrhythmia-like events and prolongation of repolarization at either maximum tested or maximal clinical exposures) categorized drugs with reasonable accuracy (area under the curve values of receiver operator curves ∼0.8). hiPSC-CM line, test site, and platform had minimal influence on drug categorization. These results demonstrate the utility of hiPSC-CMs to detect drug-induced proarrhythmic effects as part of the evolving Comprehensive In Vitro Proarrhythmia Assay paradigm.

Keywords: CiPA; comprehensive in vitro proarrhythmia assay; drug-induced ventricular arrhythmia Torsade de Pointes; hiPSC-CM; human-induced pluripotent stem cell-derived cardiomycotes; microelectrode array; voltage-sensitive dyes.

Published by Elsevier Inc.

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Conflict of interest statement

DECLARATION OF INTERESTS

The following co-authors are employed by or have declared interests in MEA or VSO platform manufacturer companies: G.S., Clyde Biosciences; D.M., Axion BioSystems; and X.Z., ACEA Biosciences. The following co-authors are employed by or have declared interests in stem cell manufacturer companies: T.K., Cellular Dynamics International; and R.K., Ncardia. G.G. is an employee of and a shareholder in the biopharmaceutical company AbbVie.

Figures

**Figure 1.. Representative Traces of Four Cellular Arrhythmia-Like Events Recorded in hiPSC-CMs**
Recorded by (left) MEA and (right) VSO platforms. The horizontal scale bar equals 1 s. We refer to type A arrhythmia as a “mild” arrhythmia-like event in the text.

**Figure 2.. EP Effects of Verapamil (Low TdP Risk) across 10 Sites (15 Site/Cell Combinations)**
Panel titles represent site number followed by a three-letter code of EP platform used (AXN, Maestro [Axion BioSystems]; CLY, CellOPTIQ [Clyde Biosciences]; ECR, CardioECR [ACEA Biosciences]; AMD, AlphaMED64 [Alpha MED Scientific]; and MCS, MEA2100 [Multichannel Systems]). Druginduced repolarization prolongation (black and gray circles for Cor.4U and iCell², correspondingly, left y axis) are shown as averaged baseline- and vehicle-controlled, Fridericia rate-corrected ddFPDc/ddAPD90c. Error bars represent SEs. The bars represent the percentage of wells in which a particular arrhythmic or quiescent event was observed (see color legend). ddFPDc/ APD90c was not calculated for the drug concentrations in which ≥50% of the wells included in the analysis were arrhythmic after drug addition. Drug concentrations (in μM and x-fold above free [unbound] clinical Cmax values) are shown in the table on the bottom of the figure, along with the concentration intervals. See also Data S1.

**Figure 3.. EP Effects of Terfenadine (Intermediate TdP Risk) across 10 Sites (15 Site/Cell Combinations)**
See Figure 2 legend. A star represents data with number of replicate wells N < 5.. See also Data S2

**Figure 4.. Effects of Dofetilide (High TdP Risk) across 10 Sites (15 Site/Cell Combinations)**
See Figure 2 legend. Stars represent missing data or data with number of replicate wells N ≤ 5. See also Data S3

**Figure 5.. Three Significant Model Predictors for Model 1 Shown for All 28 Drugs**
Each data point represents individual dataset (site/ cell type combination, 15 datasets total). (A) Predictor 1, drug-induced arrhythmia-like event at any concentration (none, no arrhythmias; type A, only arrhythmia type A; other, any other arrhythmia type: B, C, D, or any combination of ≥2 arrhythmia types). (B) Predictor 4, maximum observed drug-induced repolarization prolongation or shortening (ddFPDc or ddAPD90c) at all studied drug concentrations. (C) Predictor 7, estimated drug-induced repolarization prolongation or shortening (ddFPDc or ddAPD90c) at clinical Cmax.

**Figure 6.. Model 1 (Dichotomous Model) Prediction of a Drug’s TdP Risk Category to Be Either Low or Intermediate and High Combined Averaged from All 10 Sites (15 Cell Type/Platform Combinations)**
Error bars represent 95% confidence intervals. Red dotted line represents the 0.8 threshold discussed in the text.

**Figure 7.. Model 2 Prediction of a Drug to Fall into Low, Intermediate, or High TdP Risk Category Averaged across 10 Sites (15 Cell Type/Platform Combinations)**
Error bars represent 95% confidence intervals.

See this image and copyright information in PMC

References

1. Ando H, Yoshinaga T, Yamamoto W, Asakura K, Uda T, Taniguchi T, Ojima A, Shinkyo R, Kikuchi K, Osada T, et al. (2017). A new paradigm for drug-induced torsadogenic risk assessment using human iPS cell-derived cardiomyocytes. J. Pharmacol. Toxicol. Methods 84, 111–127. - PubMed
1. Asakura K, Hayashi S, Ojima A, Taniguchi T, Miyamoto N, Nakamori C, Nagasawa C, Kitamura T, Osada T, Honda Y, et al. (2015). Improvement of acquisition and analysis methods in multi-electrode array experiments with iPS cell-derived cardiomyocytes. J. Pharmacol. Toxicol. Methods 75, 17–26. - PubMed
1. Blinova K, Stohlman J, Vicente J, Chan D, Johannesen L, Hortigon-Vinagre MP, Zamora V, Smith G, Crumb WJ, Pang L, et al. (2017). Comprehensive translational assessment of human-induced pluripotent stem cell derived cardiomyocytes for evaluating drug-induced arrhythmias. Toxicol. Sci 155, 234–247. - PMC - PubMed
1. Clements M, and Thomas N (2014). High-throughput multi-parameter profiling of electrophysiological drug effects in human embryonic stem cell derived cardiomyocytes using multi-electrode arrays. Toxicol. Sci 140, 445–461. - PubMed
1. Colatsky T, Fermini B, Gintant G, Pierson JB, Sager P, Sekino Y, Strauss DG, and Stockbridge N (2016). The Comprehensive in Vitro Proarrhythmia Assay (CiPA) initiative - update on progress. J. Pharmacol. Toxicol. Methods 81, 15–20. - PubMed

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International Multisite Study of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes for Drug Proarrhythmic Potential Assessment

Affiliations

International Multisite Study of Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes for Drug Proarrhythmic Potential Assessment

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

Publication types

MeSH terms

Grants and funding

LinkOut - more resources

Full Text Sources

Other Literature Sources