Utilization of induced pluripotent stem cells to model the molecular network regulating congenital heart disease.
Utilization of induced pluripotent stem cells to model the molecular network regulating congenital heart disease.
复制标题
利用诱导多能干细胞来模拟调节先天性心脏病的分子网络。
DOI：
10.1093/cvr/cvab373
复制
发表时间：
2022
影响因子：
10.8
通讯作者：
Wu,JosephC
中科院分区：
文献类型：
--
作者：
Mullen,McKayMS;Wu,JosephC
先天性心脏病（CHD）是指在发育的早期阶段，心房和心室出现异常，导致房间隔缺损（ASD）、室间隔缺损或两者兼而有之。ASD是最常诊断的CHD形式之一，其典型特征是左向右分流和右心室输出量增加。1冠心病可导致慢性或急性心脏病，这取决于畸形的程度。CHD是最常见的先天性出生缺陷。尽管医疗保健的技术进步已经有所不同，但患有CHD的儿童仍然面临着显著的发病率和死亡率。2，3出生时存在的心脏畸形是儿科心血管疾病的重要组成部分，占临床相关先天性残疾的相当大的比例，每1000例活产中发生4-50例。4，5最近的进展使人们对CHD的病因有了更好的了解，使临床医生能够实施新的方法来降低受影响婴儿的发病率和死亡率。然而，迄今为止，调节心脏发育导致CHD的总体机制仍然知之甚少。心血管疾病建模中的一个强大的新范例是应用诱导多能干细胞（iPSC）及其分化的心血管细胞来开发人体生理学的体外模型。1，6 iPSC是评估特定心肌细胞亚群中多种途径（例如与心脏发育，心肌细胞功能和CHD遗传学相关的基因）高度敏感的失调的有价值的工具。7具体来说，iPSC衍生的心肌细胞（iPSC-CM）作为实验模型越来越受欢迎，因为它们是人源性的，易于获得，并且可以在体外培养数周到数月。7 Ye等人1使用iPSC-CM模型更好地阐明了GATA 4-FGF 16轴在心脏发育期间促进CHD的作用，因为它与ASD相关（图1）。GATA 4是一种参与协调心脏发育的关键转录因子。FGF 16属于成纤维细胞生长因子（FGF）家族，包含22个已知在胚胎发育中用于细胞增殖、迁移和分化的结构相关成员。6为了模拟GATA 4突变相关的ASD，将GATA 4突变的iPSC和胚胎干细胞（ESC）分化为心肌细胞（CM）。2010年，一项利用遗传分析的研究揭示了GATA 4 T280 M突变与常染色体显性遗传的家族性ASD相关。8 Ye等1在此之前的研究基础上，通过从携带GATA 4基因（T280 M）遗传性ASD突变的家族队列中产生患者特异性iPSC系（iPSC-G4 T280 M）。他们还使用CRISPR/Cas9基因组编辑方法产生了携带等基因T280 M突变的人ESC系（ESCG 4 T280 M）。1这项研究首次使用人类iPSC模型来显示GATA 4 T280 M和ASD之间的直接关系，其中GATA 4突变心肌细胞中FGF 16的过表达挽救了细胞增殖缺陷。除了使用iPSC模型揭示GATA 4和FGF 16之间的串扰外，他们还检测到GATA 4 T280 M蛋白的DNA占有率显著降低，这破坏了iPSC-G4 T280 M-CM中GATA 4靶向基因的激活或抑制。1功能缺失突变是GATA 4突变诱发冠心病等心血管疾病的直接原因。Ye等的这项研究成功表明，在GATA 4基因中携带T280 M突变的患者特异性iPSCCM。
Congenital heart disease (CHD) occurs when the atria and ventricles are abnormal during the earlier stages of development, leading to atrial septal defect (ASD), ventricular septal defect, or both. ASD is among the most frequently diagnosed forms of CHD and is typically characterized by left-to-right shunting and increased right ventricular output. 1 CHD can result in chronic or acute heart disease depending on the degree of malformation. CHD is the most commonly occurring congenital birth defect. Although technological advances in healthcare have made a difference, children afflicted with CHD are still confronted with significant morbidity and mortality. 2, 3 Cardiac malformations present at birth are a substantial component of pediatric cardiovascular disease that makes up a considerable percentage of clinically relevant congenital disabilities, occurring in 4–50 per 1000 live births. 4, 5 Recent progress has led to a better understanding of the aetiology of CHD, allowing clinicians to implement new approaches that decrease morbidity and mortality rates among affected infants. Nevertheless, to date, the overall mechanisms that regulate cardiac development leading to CHD remain poorly understood. A powerful novel paradigm in cardiovascular disease modeling is the application of induced pluripotent stem cells (iPSCs) and their differentiated cardiovascular cells to develop in vitro models of human physiology. 1, 6 iPSCs are a valuable tool for evaluating highly sensitive dysregulation of multiple pathways (eg genes associated with heart development, cardiomyocyte function, and CHD genetics) in specific subpopulations of cardiomyocytes. 7 Specifically, iPSC-derived cardiomyocytes (iPSC-CMs) have gained popularity as experimental models because they are human derived and readily available and can be cultured in vitro for weeks to months. 7 Ye et al. 1 used the iPSC-CM model to better elucidate the role of the GATA4–FGF16 axis in promoting the CHD during heart development, as it relates to ASD (Figure 1). GATA4 is a critical transcription factor involved in coordinating heart development. FGF16 belongs to the fibroblast growth factor (FGF) family, containing 22 structurally related members known for cell proliferation, migration, and differentiation in embryonic development. 6 To model the GATA4 mutation-associated ASD, both GATA4-mutant iPSCs and embryonic stem cells (ESCs) were differentiated into cardiomyocytes (CMs). In 2010, a study utilized genetic analysis to reveal that the GATA4 T280M mutation is associated with familial ASD in an autosomal dominant inheritance. 8 Ye et al. 1 built upon this previous study by generating a patient-specific iPSC line (iPSC-G4 T280M) from a family cohort carrying a hereditary ASD mutation in the GATA4 gene (T280M). They also generated a human ESC line (ESCG4 T280M) carrying the isogenic T280M mutation using the CRISPR/Cas9 genome editing method. 1 This study represents the first time that the human iPSC model was used to show the direct relationship between GATA4 T280M and ASD, in which overexpression of FGF16 in GATA4-mutant cardiomyocytes rescued the cell proliferation defect. 1 In addition to using the iPSC model to reveal the crosstalk between GATA4 and FGF16, they detected significantly lower DNA occupancy of the GATA4 T280M protein, which disrupts activation or suppression of GATA4-targeted genes in iPSC-G4 T280M-CMs. 1 Loss-of-function mutations are the direct causes of GATA4 mutation-induced cardiovascular diseases such as CHD.This study by Ye et al. successfully showed that patient-specific iPSCCMs harboring a mutation of T280M in the GATA4 …