Ambient temperature CiPA dynamic hERG protocol dataset recorded on the SyncroPatch 384SE

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. Description High-throughput patch clamp electrophysiology dataset acquired on Nanion SyncroPatch 384PE platform describing the potency and kinetics of drug block of hERG channels stably expressed in CHO cells. The voltage protocols used were 10 or 40 s 0mV steps with a 15s, -80mV inter-pulse intervals, repeated 5 or 10 times in the absence (control) and presence of drug. Drug was added and cells clamped at -80 mV for 4 minutes prior to recording to ensure drug equilibration prior to recording the onset (kinetics) of drug block. This dataset accompanies the paper titled 'High throughput measurement of hERG drug block kinetics using the CiPA dynamic protocol' by Monique J. Windley, Jessica Farr, Clifford TeBay, Jamie I. Vandenberg and Adam P. Hill. File information Raw and leak corrected files for individual plates run on the SyncroPatch 384PE Folder naming convention: Plate identifier (6 digit date_user initials_plate number)_type of data (leak uncorrected raw / leak corrected). Raw data File naming convention: Plate identifier (6 digit date_user initials_plate number)_well number Each raw data folder also contains a file named QC_plate identifier no. which contains information used for cell/patch and hERG current quality controls including seal resistance, series resistance, capacitance, baseline (average current at -80mV), average (average leak uncorrected current during last 100 ms of 0mV step). QC details are reported for each sweep along with the drug and concentration, with external and 0 M reported for control sweeps. Full SyncroPatch plates were not used for all runs, only applicable wells were included where partial plates were run (see plates 240719_MW1-3, 170719_MW1-3, 180719_MW1 and 291119_MW1) Leak corrected data File naming convention: well number_drug_concentration in M_leak_correction Refer to folder for plate identifier number. Each folder containing leak corrected data also contains a file labelled 'percent block' which contains percentage block measurements for each leak correction file in the same folder as identified by the well number. Only files passing the first pass QC were leak corrected (>125 MOhm seal, <80 MOhm series, 10-200 pA capacitance, >-300 pA baseline, >50 pA hERG current size and <20% change in peak control current)

摘要 体外促心律失常综合试验（Comprehensive in vitro Proarrhythmic Assay, CiPA）推动了药物对心脏复极影响的计算机模拟模型的应用，以优化促心律失常风险预测能力。此类模型包含描述药物结合hERG通道（human ether-à-go-go-related gene）的药效学组分，该组分除需获取阻滞效价数据外，还需要阻滞动力学的体外实验数据以实现模型约束。迄今为止，相关模型的开发与验证均基于手动膜片钳数据开展。若要规模化应用该方法，则需开发高通量自动化膜片钳（automated patch-clamp, APC）实施方案。本文针对自动化膜片钳平台上Milnes方案（或称CiPA动态方案）的实施开展了全面分析，涵盖质量控制与数据分析环节。研究针对苄普地尔、西沙必利、特非那定与维拉帕米四种药物，评估了其对hERG通道的阻滞动力学与效价；整体数据留存/质量控制（QC）通过率为21.8%，若仅针对动力学较快的药物采用适配的扫迹时长，则通过率最高可达50.4%。手动与自动化膜片钳之间的IC50与动力学变异性，与既往自动化膜片钳效价研究中不同试验站点/平台间的变异性相当。尽管本研究的实验成功率低于单纯效价筛选的结果，但仍显著高于手动膜片钳技术。通过本研究推荐的方案设计优化措施（包括扫迹时长、重复次数与漏电流校正方法），该方案可在自动化膜片钳平台上应用，以获取与手动膜片钳相当的实验数据。

数据集概述 本数据集为在Nanion SyncroPatch 384PE平台上获取的高通量膜片钳电生理数据集，描述了稳定表达于中国仓鼠卵巢（Chinese Hamster Ovary, CHO）细胞中的hERG通道的药物阻滞效价与动力学特征。所用电压刺激方案为：10 s或40 s的0 mV阶跃刺激，搭配15 s的-80 mV脉冲间期；在无药物（对照）与存在药物的条件下，分别重复5次或10次。给药后将细胞钳制于-80 mV达4分钟，以确保药物在记录药物阻滞起效（动力学）前达到平衡。本数据集配套发表于Monique J. Windley、Jessica Farr、Clifford TeBay、Jamie I. Vandenberg与Adam P. Hill合著的论文《基于CiPA动态方案的hERG药物阻滞动力学高通量测量》。

文件信息
1. 原始数据与漏电流校正文件：对应SyncroPatch 384PE平台运行的单个板孔实验数据。
文件夹命名规则：板孔标识符（6位日期_用户姓名首字母_板孔编号）_数据类型（未校正漏电流的原始数据/漏电流校正后数据）。
2. 原始数据文件命名规则：板孔标识符（6位日期_用户姓名首字母_板孔编号）_孔位编号。
每个原始数据文件夹还包含一个名为"QC_板孔标识符"的文件，其中记录了用于细胞/膜片钳与hERG电流质量控制的相关信息，包括封接电阻、串联电阻、电容、基线（-80 mV处的平均电流）、0 mV阶跃最后100 ms内未校正漏电流的平均电流。质量控制详情将随每个扫迹记录与对应药物及浓度一同上报，对照扫迹将标注为external与0 M。
3. 本次实验未使用完整的SyncroPatch板孔，仅纳入了部分可用孔位（如板孔240719_MW1-3、170719_MW1-3、180719_MW1与291119_MW1）。
4. 漏电流校正后数据文件命名规则：孔位编号_药物浓度（单位为M）_leak_correction，可参考对应文件夹获取板孔标识符。
每个包含漏电流校正后数据的文件夹还包含一个名为"percent block"的文件，其中记录了该文件夹内每个漏电流校正文件的阻滞百分比测量值，通过孔位编号进行标识。仅通过首轮质量控制的文件才会进行漏电流校正，合格标准为：封接电阻>125 MΩ、串联电阻<80 MΩ、电容介于10~200 pA、基线电流>-300 pA、hERG峰值电流>50 pA、对照峰值电流变化率<20%。

搜集汇总

背景与挑战

背景概述

该数据集是一个在Nanion SyncroPatch 384PE平台上记录的高吞吐量膜片钳电生理学数据集，专注于测量药物（如bepridil、cisapride等）对hERG通道的效力和阻断动力学，以支持CiPA（综合体外致心律失常性评估）倡议。数据集包含原始和泄漏校正数据，并提供了详细的质量控制信息，适用于药物心脏安全性评估和计算模型验证研究。