Calcium Signaling Consequences of RyR2-S4938F Mutation Expressed in Human iPSC-Derived Cardiomyocytes.
Calcium Signaling Consequences of RyR2-S4938F Mutation Expressed in Human iPSC-Derived Cardiomyocytes.
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RyR2-S4938F突变在人IPSC来源的心肌细胞中表达的钙信号后果。
DOI：
10.3390/ijms242015307
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发表时间：
2023-10-18
影响因子：
5.6
通讯作者：
中科院分区：
文献类型：
--
作者：
2型兰尼碱受体（RyR 2）是心肌肌浆网（SR）的主要钙离子释放通道，控制着心脏的节律和强度，但其功能障碍可导致严重的心律失常，导致心脏性猝死或心力衰竭。使用CRISPR/Cas9基因编辑技术在人诱导多能干细胞衍生的心肌细胞（hiPSC-CM）中表达羧基端的S4938 F-RyR 2突变。采用全内反射荧光显微镜（TIRF）和膜片钳技术研究突变基因转染的搏动心肌细胞的钙信号和电生理特性。在突变体细胞中，通过去极化至零mV或复极化从+100 mV至-50 mV测量的L型Ca 2+电流（伊卡）及其激活的Ca 2+瞬变显著较小，尽管与野生型（WT）细胞相比，其咖啡因触发的Ca 2+释放信号更大，表明ICa诱导的Ca 2+释放（CICR）受到损害。S4938 F-RyR 2细胞的更大的SR Ca 2+含量可能是自发发生的Ca 2+火花和Ca 2+瞬变及其致瘤表型的更高频率的基础。
Type-2 ryanodine receptor (RyR2) is the major Ca2+ release channel of the cardiac sarcoplasmic reticulum (SR) that controls the rhythm and strength of the heartbeat, but its malfunction may generate severe arrhythmia leading to sudden cardiac death or heart failure. S4938F-RyR2 mutation in the carboxyl-terminal was expressed in human induced pluripotent stem cells derived cardiomyocytes (hiPSC-CMs) using CRISPR/Cas9 gene-editing technique. Ca2+ signaling and electrophysiological properties of beating cardiomyocytes carrying the mutation were studied using total internal reflection fluorescence microscopy (TIRF) and patch clamp technique. In mutant cells, L-type Ca2+ currents (ICa), measured either by depolarizations to zero mV or repolarizations from +100 mV to –50 mV, and their activated Ca2+ transients were significantly smaller, despite their larger caffeine-triggered Ca2+ release signals compared to wild type (WT) cells, suggesting ICa-induced Ca2+ release (CICR) was compromised. The larger SR Ca2+ content of S4938F-RyR2 cells may underlie the higher frequency of spontaneously occurring Ca2+ sparks and Ca2+ transients and their arrhythmogenic phenotype.
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