Related Experiment Video
Updated: Sep 6, 2025
High-Throughput Cardiotoxicity Screening Using Mature Human Induced Pluripotent Stem Cell-Derived Cardiomyocyte Monolayers
Published on: March 24, 2023
High throughput measurement of hERG drug block kinetics using the CiPA dynamic protocol
Monique J Windley1, Jessica Farr2, Clifford TeBay2
1Victor Chang Cardiac Research Institute, Darlinghurst, NSW 2010, Australia; School of Clinical Medicine, Faculty of Medicine and Health, UNSW Sydney, Australia.
Insights
Automated patch-clamp successfully implements the Comprehensive in vitro Proarrhythmic Assay (CiPA) dynamic protocol for drug-induced cardiac events. This high-throughput method provides data comparable to manual patch-clamp, enhancing proarrhythmic risk prediction.
Area of Science:
- Cardiovascular Pharmacology
- Computational Toxicology
- Electrophysiology
Background:
- The Comprehensive in vitro Proarrhythmic Assay (CiPA) utilizes in silico models for drug proarrhythmic risk prediction.
- These models require in vitro data on drug interactions with hERG channels, specifically kinetics and potency of block.
- Current validation relies on manual patch-clamp, limiting scalability.
Purpose of the Study:
- To implement and analyze the CiPA dynamic protocol on a high-throughput automated patch-clamp (APC) platform.
- To assess the feasibility and data quality of APC for obtaining hERG channel block kinetics and potency.
- To compare APC performance with manual patch-clamp for CiPA protocol execution.
Main Methods:
- Implementation of the Milnes (CiPA dynamic) protocol on an APC platform.
- Assessment of drug kinetics and potency of block for bepridil, cisapride, terfenadine, and verapamil.
- Detailed quality control and data analysis procedures were applied.
Main Results:
- A data retention/QC pass rate of 21.8% was achieved overall, increasing to 50.4% with optimized sweep lengths for fast-kinetics drugs.
- Variability in IC50 and kinetics between manual and APC methods was comparable to inter-site variability in previous APC studies.
- Experimental success, while lower than potency-only screens, significantly exceeded manual patch-clamp limitations.
Conclusions:
- The CiPA dynamic protocol can be successfully implemented on an APC platform.
- Protocol modifications, including sweep length, repetitions, and leak correction, are recommended for optimal APC performance.
- APC offers a scalable solution for acquiring data comparable to manual patch-clamp, advancing in silico cardiac safety assessments.
Abstract:
The Comprehensive in vitro Proarrhythmic Assay (CiPA) has promoted use of in silico models of drug effects on cardiac repolarization to improve proarrhythmic risk prediction. These models contain a pharmacodynamic component describing drug binding to hERG channels that required in vitro data for kinetics of block, in addition to potency, to constrain them. To date, development and validation has been undertaken using data from manual patch-clamp. The application of this approach at scale requires the development of a high-throughput, automated patch-clamp (APC) implementation. Here, we present a comprehensive analysis of the implementation of the Milnes, or CiPA dynamic protocol, on an APC platform, including quality control and data analysis. Kinetics and potency of block were assessed for bepridil, cisapride, terfenadine and verapamil with data retention/QC pass rate of 21.8% overall, or as high as 50.4% when only appropriate sweep lengths were considered for drugs with faster kinetics. The variability in IC50 and kinetics between manual and APC was comparable to that seen between sites/platforms in previous APC studies of potency. Whilst the experimental success is less than observed in screens of potency alone, it is still significantly greater than manual patch. With the modifications to protocol design, including sweep length, number of repetitions, and leak correction recommended in this study, this protocol can be applied on APC to acquire data comparable to manual patch clamp.
More Related Videos
07:42Contractions of Human-iPSC-derived Cardiomyocyte Syncytia Measured with a Ca-sensitive Fluorescent Dye in Temperature-controlled 384-well Plates
Published on: October 18, 2018
10:30Technical Applications of Microelectrode Array and Patch Clamp Recordings on Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Published on: August 4, 2022
