T(brachyury)基因点突变致脊柱侧凸的分子机制研究

项目名称： T(brachyury)基因点突变致脊柱侧凸的分子机制研究

项目编号： No.81472050

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 杨雪艳

作者单位： 复旦大学

项目金额： 72万元

中文摘要： 脊柱侧凸是一种具有家族聚集性倾向的疾病，其遗传模式复杂多样。在我们研究中发现了一个包括12名患者的常染色体显性遗传脊柱侧凸家系，通过连锁分析定位其致病基因为T brachyury基因。T基因编码一种T-box家族转录因子。T基因突变可导致脊柱侧凸之前从未见报道过。但T基因集中于胚胎发育期的脊索中表达，脊索对于脊柱和体节的生成起着重要的引导作用。因此T基因突变很可能是因为对下游靶基因的调控发生变化从而引起脊柱发育异常。凝胶阻滞电泳和表面等离子共振实验均显示该家系中的T（Q199R）突变显著增强了T基因与DNA的结合能力。因此本研究拟采用CRISPR/Cas9技术通过同源重组构建T（Q199R）小鼠模型，模拟人类脊柱侧凸发病过程，通过分子、细胞、模式动物等不同水平的功能研究揭示出T基因突变导致脊柱侧凸的作用机制，深化对家族性脊柱侧凸发病机理的认识。

中文关键词： 脊柱侧凸；致病基因；基因突变；转录因子；分子机制

英文摘要： Scotiosis shows family aggregation. However its genetic patterns are complex and variable. In our study, we found a pedigree including 12 scotiosis patients, which is autosomal dominant inheritance. Linkage analysis located the pathogenic gene to T Brachyury, which is a T-box transcription factor. It is never reported that T Brachyury is related to scotiosis.T brachyury is expressed exlusively in notochord during embryo development, which play important roles in vertebrate and somites formation. Thus T gene mutation might lead to vertebral malformation via affecting regulation of its target genes. Our preliminary results via EMSA and SPR demonstrated that the binding affinity with DNA was significantly increased in T（Q199R） mutant comparing with T wildtype. In this study, we prepare to establish T（Q199R） mouse model by CRISPR/Cas9 gene editing technology, which may mimic human scotiosis disease development process. We want to clarify the mechanism of scotiosis caused by T brachyury mutation via function study.

英文关键词： scotiosis;causive gene;point mutation;trancription factor;molecular mechanism