Next-Generation Sequencing of a 40 Mb Linkage Interval Reveals TSPAN12 Mutations in Patients with Familial Exudative Vitreoretinopathy
Next-Generation Sequencing of a 40 Mb Linkage Interval Reveals TSPAN12 Mutations in Patients with Familial Exudative Vitreoretinopathy
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DOI：
10.1016/j.ajhg.2009.12.016
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发表时间：
2010-02-12
影响因子：
9.8
通讯作者：
Collin, Rob W. J.
中科院分区：
文献类型：
--
作者：
Nikopoulos, Konstantinos;Gilissen, Christian;Collin, Rob W. J.
家族性渗出性玻璃体视网膜病变（FEVR）是一种遗传异质性视网膜疾病，其特征是周边视网膜血管异常，常伴有视网膜脱离。到目前为止，已表明三个基因（FZD4、LRP5和NDP）的突变是FEVR的病因。在两个患有常染色体显性FEVR的荷兰大家系中，全基因组单核苷酸多态性（SNP）分析在7号染色体上确定了一个约40Mb的FEVR位点。微卫星标记分析表明两个家系的患者具有相似的风险单倍型。为了确定致病基因，我们对其中一个家系的先证者应用了新一代测序技术，分析了40Mb连锁区间内338个基因的所有外显子和内含子 - 外显子边界，以及微小RNA、非编码RNA和其他高度保守的基因组区域。在对序列数据进行详细的生物信息学分析后，对所有检测到的序列变异进行优先级排序，确定了三个可能是该家系致病遗传缺陷的候选基因。其中一个变异是跨膜4超家族成员12蛋白（由TSPAN12编码）中的丙氨酸到脯氨酸的替换。最近发现该蛋白与FZD4、LRP5和NDP编码的蛋白一起参与调节视网膜血管的发育。对TSPAN12的序列分析显示，在11个FEVR家系中的5个家系中存在两种分离的突变，这表明TSPAN12的突变是FEVR相对常见的病因。此外，我们证明了靶向新一代测序技术在连锁区间内鉴定疾病基因的能力。
Familial exudative vitreoretinopathy (FEVR) is a genetically heterogeneous retinal disorder characterized by abnormal vascularisation of the peripheral retina, often accompanied by retinal detachment. To date, mutations in three genes (FZD4, LRP5, and NDP) have been shown to be causative for FEVR. In two large Dutch pedigrees segregating autosomal-dominant FEVR, genome-wide SNP analysis identified an FEVR locus of similar to 40 Mb on chromosome 7. Microsatellite marker analysis suggested similar at risk haplotypes in patients of both families. To identify the causative gene, we applied next-generation sequencing in the proband of one of the families, by analyzing all exons and intron-exon boundaries of 338 genes, in addition to microRNAs, noncoding RNAs, and other highly conserved genomic regions in the 40 Mb linkage interval. After detailed bioinformatic analysis of the sequence data, prioritization of all detected sequence variants led to three candidates to be considered as the causative genetic defect in this family. One of these variants was an alanine-to-proline substitution in the transmembrane 4 superfamily member 12 protein, encoded by TSPAN12. This protein has very recently been implicated in regulating the development of retinal vasculature, together with the proteins encoded by FZD4, LRP5, and NDP. Sequence analysis of TSPAN12 revealed two mutations segregating in five of 11 FEVR families, indicating that mutations in TSPAN12 are a relatively frequent cause of FEVR. Furthermore, we demonstrate the power of targeted next-generation sequencing technology to identify disease genes in linkage intervals.