诱导多能干细胞在遗传性心肌病细胞模型中的研究进展

doi:10.3877/cma.j.issn.2095-1221.2023.05.009

遗传性心肌病是一类由于基因突变而引起的心肌结构和功能异常的疾病，可导致心力衰竭、心律失常甚至猝死。诱导多能干细胞的发现推动了人类多种疾病的体外研究，诱导获得的心肌细胞（CMs）为遗传性心肌病的机制研究及药物开发提供了重要的疾病细胞模型，已在基础研究中广泛应用。本文总结了遗传性心肌病的种类、临床表现以及常见的致病突变基因，以诱导多能干细胞衍生的CMs为体外模型，模拟各类遗传性心肌病的表型，并进一步挖掘由于基因突变引发的致病机制，为遗传性心肌病的遗传、分子和细胞学发病机制提供了充足的种子细胞，也为疾病的药物开发提供了研究模型。

关键词: 遗传性, 心肌病, 诱导多能干细胞, 细胞模型

Hereditary cardiomyopathy is a group of diseases characterized by abnormalities in myocardial structure and function due to genetic mutations, which leads to heart failure, arrhythmias and even sudden death. The discovery of induced pluripotent stem cells has promoted the in vitro study of a variety of human diseases. The cardiomyocytes obtained by induction provide important disease cell models for the study of the mechanism of hereditary cardiomyopathies and drug development and have been widely used in basic research. This article summarizs the types, clinical manifestations and common pathogenic mutated genes of hereditary cardiomyopathy, used induced pluripotent stem cell-derived cardiomyocytes as in vitro models to simulate the phenotypes of various types of hereditary cardiomyopathy, and further explores the related pathogenic mechanisms caused by gene mutations, providing sufficient seed cells for the genetic, molecular and cytological pathogenesis of hereditary cardiomyopathy. It also provides a valuable research model for drug development of diseases.

Key words: Hereditary, Cardiomyopathy, Induced pluripotent stem cells, Cell model

图1 心肌病患者来源的iPSCs用于心肌病细胞模型的研究路线注：iPSCs为人诱导多能干细胞

表1 hiPSC在遗传性心肌病细胞模型中的应用

疾病名称	发表年份	亮点	参考文献
HCM	2013	探索MYH7基因（p.R663H）突变在HCM中的作用机制，基于hiPSC模型发现与对照组相比，HCM组具有细胞增大和收缩性心律失常及细胞内钙循环增多等表型。	[22]
	2019	HCM患者hiPSC-CMs表现出舒张功能受损，表现为舒张时间延长、舒张率降低和舒张肌节长度缩短以及舒张期钙超载等表型。	[23]
	2022	首次证明iPSC-CMs中存在功能性的MPO，且在HCM-CMs中上调，通过增加3-氯酪氨酸修饰的MYBPC3蛋白水平来降低MYBPC3磷酸化水平，进而损害HCM-CMs中的心肌细胞舒张动力学。	[24]
	2018	PRKAG2突变的HCM hiPSC-CMs表现出结构和功能异常。利用CRISPR矫正后消除了电生理异常、糖原累积和心肌细胞肥大等异常表型。	[25]
DCM	2012	首次建立了携带TNNT2突变的DCM-hiPSC疾病模型。	[31]
	2015	利用TNNT2突变患者的hiPSC-CMs表征了心肌细胞在体外分化和成熟过程中的特性，并发现β肾上腺素能信号通路传导功能在DCM hiPSC-CMs中被减弱。	[32]
	2020	利用TNNT2突变的hiPSC系，揭示了基因突变后破坏了肌钙蛋白与其他肌节蛋白的相互作用。	[33]
LVNC	2023	利用KLHL26突变患者衍生出hiPSC-CMs，细胞表现出内质网扩张和线粒体畸形，以及收缩率降低、钙瞬变等异常表型。	[38]
	2023	利用PLEKHM2突变患者衍生hiPSC-CMs，发现这些心肌细胞自噬活性水平较低。	[39]
	2016	构建TBX20突变的LVNC患者的hiPSC-CMs，发现TBX20突变后TGFβ信号转导异位激活，最终损害了心肌多层致密结构的形成。利用CRISPR技术矫正后，降低了TGFβ信号转导活性。	[40]
ARVC	2020	利用OBSCN突变的ARVC-hiPSC生成心肌细胞，表现出明显的脂质积累、多形性增加、不规则的Z带和L型钙电流增加。	[44]
	2020	利用PKP2-hiPSC-CMs模型再现了ARVC脂肪生成和细胞凋亡的表型，并发现加入睾酮后表型恶化，雌二醇处理后表型有所改善。	[45]
	2013	利用PKP2突变的ARVC患者hiPSC诱导为具有异常JUP蛋白核易位的心肌细胞，且脂肪生成、细胞凋亡、活性氧产生和脂肪酸氧化通量等方面均显著增加，PPARγ拮抗剂可减少脂肪生成，添加活性氧清除剂可减少心肌细胞凋亡。	[46]
	2018	利用DSG2突变的ARVC患者的hiPSC生成心肌细胞，表现出多离子通道功能障碍和异常的细胞电生理学，以及对肾上腺素能刺激的敏感性增强。	[47]

表1 hiPSC在遗传性心肌病细胞模型中的应用

疾病名称	发表年份	亮点	参考文献
HCM	2013	探索MYH7基因（p.R663H）突变在HCM中的作用机制，基于hiPSC模型发现与对照组相比，HCM组具有细胞增大和收缩性心律失常及细胞内钙循环增多等表型。	[22]
	2019	HCM患者hiPSC-CMs表现出舒张功能受损，表现为舒张时间延长、舒张率降低和舒张肌节长度缩短以及舒张期钙超载等表型。	[23]
	2022	首次证明iPSC-CMs中存在功能性的MPO，且在HCM-CMs中上调，通过增加3-氯酪氨酸修饰的MYBPC3蛋白水平来降低MYBPC3磷酸化水平，进而损害HCM-CMs中的心肌细胞舒张动力学。	[24]
	2018	PRKAG2突变的HCM hiPSC-CMs表现出结构和功能异常。利用CRISPR矫正后消除了电生理异常、糖原累积和心肌细胞肥大等异常表型。	[25]
DCM	2012	首次建立了携带TNNT2突变的DCM-hiPSC疾病模型。	[31]
	2015	利用TNNT2突变患者的hiPSC-CMs表征了心肌细胞在体外分化和成熟过程中的特性，并发现β肾上腺素能信号通路传导功能在DCM hiPSC-CMs中被减弱。	[32]
	2020	利用TNNT2突变的hiPSC系，揭示了基因突变后破坏了肌钙蛋白与其他肌节蛋白的相互作用。	[33]
LVNC	2023	利用KLHL26突变患者衍生出hiPSC-CMs，细胞表现出内质网扩张和线粒体畸形，以及收缩率降低、钙瞬变等异常表型。	[38]
	2023	利用PLEKHM2突变患者衍生hiPSC-CMs，发现这些心肌细胞自噬活性水平较低。	[39]
	2016	构建TBX20突变的LVNC患者的hiPSC-CMs，发现TBX20突变后TGFβ信号转导异位激活，最终损害了心肌多层致密结构的形成。利用CRISPR技术矫正后，降低了TGFβ信号转导活性。	[40]
ARVC	2020	利用OBSCN突变的ARVC-hiPSC生成心肌细胞，表现出明显的脂质积累、多形性增加、不规则的Z带和L型钙电流增加。	[44]
	2020	利用PKP2-hiPSC-CMs模型再现了ARVC脂肪生成和细胞凋亡的表型，并发现加入睾酮后表型恶化，雌二醇处理后表型有所改善。	[45]
	2013	利用PKP2突变的ARVC患者hiPSC诱导为具有异常JUP蛋白核易位的心肌细胞，且脂肪生成、细胞凋亡、活性氧产生和脂肪酸氧化通量等方面均显著增加，PPARγ拮抗剂可减少脂肪生成，添加活性氧清除剂可减少心肌细胞凋亡。	[46]
	2018	利用DSG2突变的ARVC患者的hiPSC生成心肌细胞，表现出多离子通道功能障碍和异常的细胞电生理学，以及对肾上腺素能刺激的敏感性增强。	[47]

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Research progress of induced pluripotent stem cells in cell models of hereditary cardiomyopathy

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Research progress of induced pluripotent stem cells in cell models of hereditary cardiomyopathy