SMYD4：一个新的先天性心脏病候选基因的识别与验证

先天性心脏病（简称先心病）是严重影响患者生命和生活质量的出生缺陷，研究其发病机制有重要意义。我们前期的全外显子组测序结果提示，SMYD4可能是新的先心病候选基因，但需进一步证实。拟开展以下研究：①检测SMYD4基因表达、DNA序列及其表达调控，揭示先心病患者中是否存在SMYD4基因缺陷；②采用CRISPR/Cas9技术敲除模式生物斑马鱼smyd4基因，明确smyd4基因缺陷对斑马鱼心脏表型的影响；③在细胞水平研究SMYD4基因对心肌细胞功能的影响，分析SMYD4基因相互作用蛋白，明确SMYD4在心脏发育过程中所参与的信号通路/调节网络以及SMYD4参与先心病发生的机制。通过上述系列研究，在先心病人群、模式生物斑马鱼和模式细胞等多种水平，从表型到基因型、再从基因型到表型，并进行机制研究，逐层深入地阐明SMYD4基因在先心病发生中的作用及其机制。该研究思路可为其他出生缺陷的病因学研究提供参考。

Congenital heart disease (CHD), one of the most common birth defect, affects the life quality of the patients and is also the leading cause of death in infants. So,it’s important to study the pathogenesis of CHD. Our previous whole exome sequencing (WES) results have suggested that the gene SMDY4 may be a new candidate gene of CHD. But this phenomenon and its mechanism should be further confirmed. We plan to launch the following research, including ① Detecting the DNA sequence, the expression and its regulation of the gene SMYD4, to reveal whether there’re the SMYD4 gene defects in patients with CHD. ② To create the model organism SMDY4 gene knockout zebrafish by using the technique of CRISPR/Cas9, then elucidate the cardiac phenotype of the zebrafish with SMYD4 gene defect. ③ To make clear the effect of SMYD4 gene on the function of myocardial cell, analyze the proteins interacting with SMYD4, and explicate the signal pathway or regulation network in which the gene SMYD4 participating at the cellular level, and the molecular mechanism of SMYD4 in the occurrence of CHD. Based on the above serial study, on the level of patients with CHD, model organism zebrafish and model cell, from phenotype to genotype, then from genotype to phenotype, combined with the mechanistic investigation, we expect to elucidate the role of gene SMYD4 in the pathogenesis of CHD and its mechanism. In addition, the research thinking and method will provide references for other birth defects.