Wang Feng, Liu Dong, Zhang Ran-Ran, Yu Li-Wei, Zhao Jian-Yuan, Yang Xue-Yan, Jiang Song-Shan, Ma Duan, Qiao Bin, Zhang Feng, Jin Li, Gui Yong-Hao, Wang Hong-Yan
Children's Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Institute of Reproduction & Development, Fudan University, Shanghai, China.
Co-innovation Center of Neuroregeneration, Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Nantong University, Nantong, China.
Cell Discov. 2017 Jul 25;3:17026. doi: 10.1038/celldisc.2017.26. eCollection 2017.
TBX5 is a vital transcription factor involved in cardiac development in a dosage-dependent manner. But little is known about the potential association of 3' untranslated region (UTR) variations with congenital cardiac malformations. This study aimed to investigate the relationship between 3'UTR variants and risk for congenital heart disease (CHD) susceptibility in two Han Chinese populations, and to reveal its molecular mechanism. The relationship between 3'UTR variants and CHD susceptibility was examined in 1 177 CHD patients and 990 healthy controls in two independent case-control studies. Variant rs6489956 C>T was found to be associated with increased CHD susceptibility in both cohorts. The combined CHD risk for the CT and TT genotype carriers was 1.83 times higher than that of CC genotype, while the risk for CT or TT genotype was 1.94 times and 2.31 times higher than that of CC carriers, respectively. Quantitative real-time PCR and western blot analysis showed that T allele carriers exhibited reduced mRNA and protein levels in CHDs tissues. Compared with C allele, T allele showed increased binding affinity to miR-9 and miR-30a in both luciferase assays and surface plasmon resonance analysis. Functional analysis confirmed that miR-9 and miR-30a downregulated expression at the transcriptional and translational levels, respectively. The assays in zebrafish model were in support of the interaction of miR-9/30a and TBX5 3'UTR (C and T allele). We concluded that 3'UTR variant rs6489956 increased susceptibility of CHD in the Han Chinese population because it changes the binding affinity of two target miRNAs that specifically mediate expression.
TBX5是一种重要的转录因子，以剂量依赖的方式参与心脏发育。但关于3'非翻译区（UTR）变异与先天性心脏畸形的潜在关联知之甚少。本研究旨在探讨两个汉族人群中3'UTR变异与先天性心脏病（CHD）易感性风险之间的关系，并揭示其分子机制。在两项独立的病例对照研究中，对1177例CHD患者和990例健康对照进行了3'UTR变异与CHD易感性关系的研究。发现变异rs6489956 C>T在两个队列中均与CHD易感性增加相关。CT和TT基因型携带者的CHD综合风险比CC基因型高1.83倍，而CT或TT基因型的风险分别比CC携带者高1.94倍和2.31倍。定量实时PCR和蛋白质印迹分析表明，T等位基因携带者在CHD组织中的mRNA和蛋白质水平降低。在荧光素酶测定和表面等离子体共振分析中，与C等位基因相比，T等位基因显示出与miR-9和miR-30a的结合亲和力增加。功能分析证实，miR-9和miR-30a分别在转录和翻译水平下调表达。斑马鱼模型试验支持miR-9/30a与TBX5 3'UTR（C和T等位基因）的相互作用。我们得出结论，3'UTR变异rs6489956增加了汉族人群中CHD的易感性，因为它改变了两个特异性介导表达的靶miRNA的结合亲和力。
