Department of Chemistry, Stanford University, Stanford, CA, 94305, USA.
Stanford Cardiovascular Institute, Stanford University, Stanford, CA, USA.
Biosens Bioelectron. 2022 Nov 15;216:114617. doi: 10.1016/j.bios.2022.114617. Epub 2022 Aug 13.
Unintended binding of small-molecule drugs to ion channels affects electrophysiological properties of cardiomyocytes and potentially leads to arrhythmia and heart failure. The waveforms of intracellular action potentials reflect the coordinated activities of cardiac ion channels and serve as a reliable means for assessing drug toxicity, but the implementation is limited by the low throughput of patch clamp for intracellular recording measurements. In the last decade, several new technologies are being developed to address this challenge. We recently developed the nanocrown electrode array (NcEA) technology that allows robust, parallel, and long-duration recording of intracellular action potentials (iAPs). Here, we demonstrate that NcEAs allow comparison of iAP waveforms before and after drug treatment from the same cell. This self-referencing comparison not only shows distinct drug effects of sodium, potassium, and calcium blockers, but also reveals subtle differences among three subclasses of sodium channel blockers with sub-millisecond accuracy. Furthermore, self-referencing comparison unveils heterogeneous drug responses among different cells. In our study, whole-panel simultaneous intracellular recording can be reliably achieved with ∼94% success rate. The average duration of intracellular recording is ∼30 min and some last longer than 2 h. With its high reliability, long recording duration, and easy-to-use nature, NcEA would be useful for iAP-based preclinical drug screening.
小分子药物与离子通道的非预期结合会影响心肌细胞的电生理特性，并可能导致心律失常和心力衰竭。细胞内动作电位的波形反映了心脏离子通道的协调活动，是评估药物毒性的可靠手段，但由于细胞内记录测量的膜片钳高通量较低，其实施受到限制。在过去的十年中，开发了几种新技术来应对这一挑战。我们最近开发了纳米冠电极阵列（NcEA）技术，该技术允许对细胞内动作电位（iAP）进行稳健、平行和长时间的记录。在这里，我们证明 NcEAs 允许从同一细胞中比较药物处理前后的 iAP 波形。这种自我参照比较不仅显示了钠、钾和钙通道阻滞剂的明显药物作用，而且还以亚毫秒级的精度揭示了三种钠通道阻滞剂亚类之间的细微差异。此外，自我参照比较揭示了不同细胞之间药物反应的异质性。在我们的研究中，整个面板的同时细胞内记录可以可靠地实现，成功率约为 94%。细胞内记录的平均持续时间约为 30 分钟，有些记录持续时间超过 2 小时。NcEA 具有高可靠性、长记录时间和易于使用的特点，将有助于基于 iAP 的临床前药物筛选。
