Cardiovascular Development and Congenital Heart Disease Modeling in the Pig.
Cardiovascular Development and Congenital Heart Disease Modeling in the Pig.
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DOI：
10.1161/jaha.121.021631
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发表时间：
2021-07-20
影响因子：
5.4
通讯作者：
Lo CW
中科院分区：
文献类型：
--
作者：
Gabriel GC;Devine W;Redel BK;Whitworth KM;Samuel M;Spate LD;Cecil RF;Prather RS;Wu Y;Wells KD;Lo CW
在小鼠身上建立心血管疾病模型为了解先天性心脏病的病因提供了宝贵的见解。然而，小鼠心脏的小尺寸阻碍了转化研究。鉴于目前的高效基因编辑，在其他物种中建立先天性心脏病模型是可能的。猪是有利的，因为它的心脏解剖，生理和大小与人类婴儿相似。我们分析了猪的心血管发育情况，并产生了带有先天性心脏缺陷的基因编辑猪。收集7期（第20天至第115天出生）的猪胚胎和胎儿，每期至少分析3个胚胎。结合磁共振成像和三维组织重建与主教共聚焦显微镜进行。对晚期或足月胎儿进行大体解剖，采用序贯的心房、心室和动脉段分析。在第20天，心脏已形成环，形成共同的心房、心室和不分离的流出道。心脏形态发生进展迅速，第26天房间隔和流出间隔明显，第30天室间隔完成。流出液和房室缓冲在第20天发生重塑形成成熟瓣膜，这一过程持续到第42天。由染色质修饰子SAP130突变产生的基因编辑猪表现出三尖瓣发育不良，三尖瓣闭锁与早期胚胎致死有关。猪心脏形态发生的主要事件在第30天基本完成。其发育特征与人类和小鼠相似，这表明基因编辑猪可能为专注于改善先天性心脏病结局的临床前研究提供新的机会。
Modeling cardiovascular diseases in mice has provided invaluable insights into the cause of congenital heart disease. However, the small size of the mouse heart has precluded translational studies. Given current high‐efficiency gene editing, congenital heart disease modeling in other species is possible. The pig is advantageous given its cardiac anatomy, physiology, and size are similar to human infants. We profiled pig cardiovascular development and generated genetically edited pigs with congenital heart defects. Pig conceptuses and fetuses were collected spanning 7 stages (day 20 to birth at day 115), with at least 3 embryos analyzed per stage. A combination of magnetic resonance imaging and 3‐dimensional histological reconstructions with episcopic confocal microscopy were conducted. Gross dissections were performed in late‐stage or term fetuses by using sequential segmental analysis of the atrial, ventricular, and arterial segments. At day 20, the heart has looped, forming a common atria and ventricle and an undivided outflow tract. Cardiac morphogenesis progressed rapidly, with atrial and outflow septation evident by day 26 and ventricular septation completed by day 30. The outflow and atrioventricular cushions seen at day 20 undergo remodeling to form mature valves, a process continuing beyond day 42. Genetically edited pigs generated with mutation in chromatin modifier SAP130 exhibited tricuspid dysplasia, with tricuspid atresia associated with early embryonic lethality. The major events in pig cardiac morphogenesis are largely complete by day 30. The developmental profile is similar to human and mouse, indicating gene edited pigs may provide new opportunities for preclinical studies focused on outcome improvements for congenital heart disease.