Identification of FOXH1 mutations in patients with sporadic conotruncal heart defect
Identification of FOXH1 mutations in patients with sporadic conotruncal heart defect
复制标题
散发性圆锥干心脏缺陷患者 FOXH1 突变的鉴定
DOI：
10.1111/cge.13710
复制
发表时间：
2020
影响因子：
3.5
通讯作者：
Xu Rang
中科院分区：
文献类型：
--
作者：
Wei Wei;Li Bojian;Li Fen;Sun Kun;Jiang Xuechao;Xu Rang
圆锥干心脏缺陷(CTD)是由于心脏流出道(OFT)发育异常而导致的先天性心脏病的一个重要亚型。FOXH1是一种转录因子，参与右心室和OFT的形态发生。在本研究中，我们证实了FOXH1在OFT发育过程中在小鼠和人类胚胎中的表达。我们还扫描了605例散发性CTD患者和300例正常对照的FOXH1基因编码外显子和剪接区，从中我们发现了7个FOXH1基因杂合性突变。根据生物信息学分析结果，预测它们在保守的氨基酸位点具有潜在的危害性。Western印迹分析表明，所有突变体均降低FOXH1蛋白的表达，而双荧光素酶报告分析表明，除p.P35R外，其余6个突变体都增强了调控MEF2C表达的能力，MEF2C与NKX2.5相互作用，参与心脏生长。凝胶迁移率改变分析结果表明，两个突变改变了突变的FOXH1蛋白的DNA结合能力。在相同突变的患者中发现表型异质性。这些结果表明FOXH1突变导致致病的功能改变，从而导致CTD的发生。
Conotruncal heart defects (CTD) are an important subtype of congenital heart disease that occur due to abnormality in the development of the cardiac outflow tract (OFT). FOXH1 is a transcription factor that participates in the morphogenesis of the right ventricle and OFT. In this study, we confirmed the expression of FOXH1 in mouse and human embryos during OFT development. We also scanned the coding exons and splicing regions of the FOXH1 gene in 605 patients with sporadic CTD and 300 unaffected controls, from which we identified seven heterozygous FOXH1 gene mutations. According to bioinformatics analysis results, they were predicted potentially deleterious at conserved amino acid sites. Western blot was used to show that all the variants decreased the expression of FOXH1 protein, while dual‐luciferase reporter assay showed that six of them, with an exception of p.P35R, had enhanced abilities to modulate the expression of MEF2C, which interacts with NKX2.5 and is involved in cardiac growth. The electrophoretic mobility shift assays result showed that two mutations altered DNA‐binding abilities of mutant FOXH1 proteins. Phenotype heterogeneity was found in patients with the same mutation. These results indicate that FOXH1 mutations lead to disease‐causing functional changes that contribute to the occurrence of CTD.