Variability in high-throughput ion-channel screening data and consequences for cardiac safety assessment

Introduction: Unwanted drug interactions with ionic currents in the heart can lead to an increased proarrhythmic risk to patients in the clinic. It is therefore a priority for safety pharmacology teams to detect block of cardiac ion channels, and new technologies have enabled the development of automated and high-throughput screening assays using cell lines. As a result of screening multiple ion-channels there is a need to integrate information, particularly for compounds affecting more than one current, and mathematical electrophysiology in-silico action potential models are beginning to be used for this. Methods: We quantified the variability associated with concentration-effect curves fitted to recordings from high-throughput Molecular Devices IonWorks (R) Quattro (TM) screens when detecting block of I-Kr (hERG), I-Na (NaV1.5), I-CaL (CaV1.2), I-Ks (KCNQ1/minK) and I-to (Kv4.3/KChIP2.2), and the Molecular Devices FLIPR (R) Tetra fluorescence screen for I-CaL (CaV1.2), for control compounds used at AstraZeneca and GlaxoSmithKline. We examined how screening variability propagates through in-silico action potential models for whole cell electrical behaviour, and how confidence intervals on model predictions can be estimated with repeated simulations. Results: There are significant levels of variability associated with high-throughput ion channel electrophysiology screens. This variability is of a similar magnitude for different cardiac ion currents and different compounds. Uncertainty in the Hill coefficients of reported concentration-effect curves is particularly high. Depending on a compound's ion channel blocking profile, the uncertainty introduced into whole-cell predictions can become significant. Discussion: Our technique allows confidence intervals to be placed on computational model predictions that are based on high-throughput ion channel screens. This allows us to suggest when repeated screens should be performed to reduce uncertainty in a compound's action to acceptable levels, to allow a meaningful interpretation of the data. (C) 2013 The Authors. Published by Elsevier Inc. All rights reserved.