Electrophysiology data for NAA10-R4S-induced pluripotent-derived cardiomyocytes

N-terminal acetyltransferases including NAA10 catalyze N-terminal acetylation, an evolutionarily conserved co- and post-translational modification. However, little is known about the role of N-terminal acetylation in cardiac homeostasis. To gain insight into cardiac-dependent NAA10 function, we studied a previously unidentified NAA10 variant p.(Arg4Ser) segregating with QT-prolongation, cardiomyopathy, and developmental delay in a large kindred. Here, we show that the NAA10R4S variant reduced enzymatic activity, decreased NAA10-NAA15 complex formation, and destabilized the enzymatic complex N-terminal acetyltransferase A. In NAA10R4S/Y-induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CMs), dysregulation of the late sodium and slow delayed rectifier potassium currents caused severe repolarization abnormalities, consistent with clinical QT prolongation. Engineered heart tissues generated from NAA10R4S/Y-iPSC-CMs had significantly decreased contractile force and sarcomeric disorg..., Whole-cell patch clamp recordings Cultured iPSC-CMs were dissociated with Accutase and plated sparsely onto Geltrex-coated 11mm coverslips. Single iPSC-CMs were analyzed 3 to 6 days after dissociating. Single iPSC-CMs were recorded under different conditions to acquire each parameter (Supplemental Table 1)50,51. Perforated patch recordings were performed for action potential (AP) analysis and the L-type Ca2+ current (ICaL). Perforated patch was applied in order to prevent run-down in ICaL recording52. The ruptured patch technique was used for INa, IKs, and IKr recordings. Series resistance and cell capacitance were compensated to ~ 60 % for all the voltage clamp experiments. To measure INa, starting from a holding potential of -100 mV, 40 ms of depolarizing pulses from -100 mV to 90 mV were applied in 10-mV increments. For the INa steady-state inactivation, following 400 ms of prepulses with 10-mV increments from -110 mV to -20 mV, 40 ms of 0 mV pulse was applied.  For IKs, test pu..., , # **Electrophysiology data for NAA10-R4S-induced pluripotent-derived cardiomyocytes** [https://doi.org/10.5061/dryad.280gb5mvz](https://doi.org/10.5061/dryad.280gb5mvz) The enclosed dataset contains electrophysiology data from patient-derived and genome-edited induced pluripotent stem cell derived cardiomyocytes (iPSC-CMs) with dysfunctional NAA10. Data includes multi-electrode array (MEA) recordings of monolayer cultures and patch-clamp electrophysiology of single cells. ### **Description of the data and file structure** Files are in the GraphPad Prism format version 9 and version 10. Data is organized by related experiment and separated by patient-derived, genome-editing and wild type (WT) lines. All files are also uploaded in .csv format. Filenames are the same for both file formats. Any empty cells are replaced with “null” in the .csv files. These empty cells were left in the prism files to facilitate statistical calculations. Description of individual files and corresponding ...,N端乙酰转移酶（N-terminal acetyltransferase）包括NAA10，可催化N端乙酰化——这是一种在进化上保守的共翻译与翻译后修饰。然而，目前对于N端乙酰化在心脏稳态中的作用仍知之甚少。 为深入探究依赖于心脏的NAA10功能，我们对一个大型家系中与QT间期延长、心肌病及发育迟缓共分离的、此前未被报道的NAA10变异体p.(Arg4Ser)展开了研究。本研究发现，NAA10R4S变异体可降低酶活性、削弱NAA10-NAA15复合物的形成，并使N端乙酰转移酶A（N-terminal acetyltransferase A）这一酶复合物的稳定性下降。 在NAA10R4S/Y诱导多能干细胞衍生心肌细胞（induced pluripotent stem cell-derived cardiomyocytes, iPSC-CMs）中，晚钠电流与缓慢延迟整流钾电流的失调会引发严重的复极化异常，这与临床观察到的QT间期延长相符。由NAA10R4S/Y-iPSC-CMs构建的工程化心脏组织，其收缩力显著下降且肌节结构紊乱…… 全细胞膜片钳记录 培养的iPSC-CMs采用Accutase消化解离，并以低密度接种于包被了Geltrex的11mm盖玻片上。解离后3至6天对单个iPSC-CMs进行分析。通过不同实验条件下的记录获取各参数（补充表1）[50,51]。采用穿孔膜片钳技术记录动作电位（action potential, AP）与L型钙电流（L-type Ca²⁺ current, ICaL），以避免ICaL记录过程中的电流衰减[52]。而INa、IKs及IKr的记录则采用破膜膜片钳技术。所有电压钳实验均将串联电阻与细胞电容补偿至约60%。为记录INa，从-100 mV的钳制电位开始，以10 mV为步长施加从-100 mV至90 mV的40 ms去极化脉冲。针对INa的稳态失活特性，先以10 mV为步长施加从-110 mV至-20 mV的400 ms预脉冲，随后施加40 ms的0 mV测试脉冲。针对IKs的测试脉冲…… # **NAA10-R4S诱导多能干细胞衍生心肌细胞电生理数据集** [https://doi.org/10.5061/dryad.280gb5mvz] 本附带数据集包含来自患者来源及基因编辑的、功能异常的NAA10诱导多能干细胞衍生心肌细胞（iPSC-CMs）的电生理数据。数据涵盖单层培养细胞的多电极阵列（multi-electrode array, MEA）记录结果，以及单个细胞的膜片钳电生理数据。 ### 数据与文件结构说明 文件采用GraphPad Prism 9及10版本格式存储。数据按相关实验分组，并按患者来源细胞系、基因编辑细胞系与野生型（wild type, WT）细胞系进行区分。所有文件同时以.csv格式上传，两种格式的文件名完全一致。.csv文件中的空单元格均替换为"null"，而Prism文件中保留空单元格以方便统计计算。 单个文件及对应……的说明