The Functional Polymorphism R129W in the BVES Gene Is Associated with Sporadic Tetralogy of Fallot in the Han Chinese Population
The Functional Polymorphism R129W in the BVES Gene Is Associated with Sporadic Tetralogy of Fallot in the Han Chinese Population
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BVES基因R129W功能多态性与汉族人群散发性法洛四联症相关
DOI：
10.1089/gtmb.2019.0085
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发表时间：
2019
影响因子：
1.4
通讯作者：
Ping
中科院分区：
文献类型：
--
作者：
Yan Shi;Yongqing Li;Yuequn Wang;Jian Zhuang;Heng Wang;Min Hu;Xiaoyang Mo;Shusheng Yue;Yu Chen;Xiongwei Fan;Jimei Chen;Wanwan Cai;Xiaolan Zhu;Yongqi Wan;Ying Zhong;Xiangli Ye;Fang Li;Zuoqiong Zhou;Guo Dai;Rong Luo;Karen Ocorr;Zhigang Jiang;Xiaoping Li;Ping
背景：法洛四联症 (TOF) 约占先天性心脏病病例的 10%。据报道，血管心外膜物质（BVES）基因在成人心脏功能中发挥作用。然而，BVES中的等位基因变异是否会导致TOF风险及其可能的机制仍不清楚。方法：使用来自146名TOF患者和100名无关健康对照的样本对BVES基因的开放阅读框进行测序。 qRT-PCR 和蛋白质印迹测定用于确认 TOF 样品中突变 BVES 变体的表达。使用在线软件 Polyphen2 和 SIFT 来预测观察到的等位基因变异的有害影响。使用双荧光报告基因测定检查这些等位基因变体对基因转录活性的影响。结果：我们对 146 名 TOF 患者中每人的 BVES 基因中的 4 个单核苷酸多态性 (SNP) 进行了基因分型。其中，c.385C>T (p.R129W) 的次要等位基因频率在 TOF 中为 0.035%，但在 100 个对照和中国百万富翁数据库中为 ∼0.025%。 Polyphen2 和 SIFT 软件预测该等位基因变异是潜在有害的改变。 qRT-PCR和蛋白质印迹分析表明，带有c.385C>T等位基因变异的6个右心室流出道样本中BVES的表达显着下调。双荧光报告系统显示c.385C>T等位变体显着降低了BVES基因的转录活性，并且还降低了GATA4和NKX2.5启动子的转录。结论：c.385C>T（p.R129W）是BVES基因的功能性SNP，可降低BVES体外和体内的转录活性 TOF 组织。这随后影响了与TOF相关的GATA4和NKX2.5的转录活性。这些发现表明，c.385C>T 可能与中国汉族人群中 TOF 的风险相关。
Background:Tetralogy of Fallot (TOF) accounts for ∼10% of congenital heart disease cases. The blood vessel epicardial substance (BVES) gene has been reported to play a role in the function of adult hearts. However, whether allelic variants inBVEScontribute to the risk of TOF and its possible mechanism remains unknown.Methods:The open reading frame of theBVESgene was sequenced using samples from 146 TOF patients and 100 unrelated healthy controls. qRT-PCR and western blot assays were used to confirm the expression of mutatedBVESvariants in the TOF samples. The online software Polyphen2 and SIFT were used to predict the deleterious effects of the observed allelic variants. The effects of these allelic variants on the transcriptional activities of genes were examined using dual-fluorescence reporter assays.Results:We genotyped four single nucleotide polymorphisms (SNPs) in theBVESgene from each of the 146 TOF patients. Among them, the minor allelic frequencies of c.385C>T (p.R129W) were 0.035% in TOF, but ∼0.025% in 100 controls and the Chinese Millionome Database. This allelic variant was predicted to be a potentially harmful alteration by the Polyphen2 and SIFT softwares. qRT-PCR and western blot analyses indicated that the expression ofBVESin the six right ventricular outflow tract samples with the c.385C>T allelic variant was significantly downregulated. A dual-fluorescence reporter system showed that the c.385C>T allelic variant significantly decreased the transcriptional activity of theBVESgene and also decreased transcription from theGATA4andNKX2.5promoters.Conclusions:c.385C>T (p.R129W) is a functional SNP of theBVESgene that reduces the transcriptional activity ofBVES in vitroandin vivoin TOF tissues. This subsequently affects the transcriptional activities ofGATA4andNKX2.5related to TOF. These findings suggest that c.385C>T may be associated with the risk of TOF in the Han Chinese population.