图雷特综合征与BDNF基因突变研究  

Association study on BDNF gene mutations and Tourette syndrome

作　　者：王炳辉 孟玉洁 刘文淼 吴云安 刘世国[3] 董继承[2] WANG Binghui;MENG Yujie;LIU Wenmiao(Women and Children’s Hospital,Qingdao University,Qingdao 266001,China)

机构地区：[1]青岛大学附属妇女儿童医院,山东青岛266001 [2]青岛市精神卫生中心 [3]青岛大学附属医院 [4]青岛安宁心理医院

出　　处：《精神医学杂志》2024年第5期506-511,共6页Journal of Psychiatry

基　　金：国家自然科学基金项目(编号:81371499,82001441);青岛市卫计委医药科研指导计划(编号:2016-WJZD067)。

摘　　要：目的对图雷特综合征(TS)患者脑源性神经营养因子(BDNF)基因进行遗传学分析,找出新的罕见致病基因突变位点。方法收集524例TS患者血液标本,提取外周静脉血全基因组DNA,利用二代靶向高通量测序技术对BDNF基因进行全部外显子检测,与人类基因组突变数据库中标准序列比对,找出候选变异位点并进行Sanger测序验证。利用生物信息学软件分析候选变异位点的致病性并预测变异蛋白的致病性,同时使用3D蛋白模型模拟变异前后蛋白质结构及分子间作用力的变化,初步确定可疑致病突变位点。结果高通量测序结果显示TS BDNF基因存在2处错义突变c.118C>T/p.H40Y、c.196G>A/p.V66M和1处缺失突变c.99_100del:p.C34Ffs*12,其中c.118C>T为尚未报道的罕见变异。生物信息学分析软件预测3个突变均为有害突变。多物种蛋白同源比对显示H40、V66位于高度保守区域;MAESTROweb预测p.H40Y降低蛋白质的稳定性;Pymol预测显示p.V66M变异使氨基酸之间的氢键发生改变,进而影响蛋白空间结构的稳定性。结论BDNF基因c.118C>T/p.H40Y、c.196G>A/p.V66M错义突变c.99_100del:p.C34Ffs*12缺失突变很可能是TS的致病突变位点。本研究有利于扩大BDNF的突变谱,为TS的治疗提供新靶点。Objective To identify the pathogenic gene variants through genetic analysis of the brain-derived neurotrophic factor(BDNF)gene in patients with Tourette syndrome(TS).Methods Whole genomic DNA was extracted from the peripheral venous blood of 524 patients with TS.Whole exome sequencing of the BDNF gene was performed using second-generation targeted high-throughput sequencing technology.Sequence data were compared with standard sequences from the Human Genome Mutation Database to identify candidate variants and verified by Sanger sequencing.Bioinformatic software was used to analyze the pathogenicity of candidate mutations.In the meantime,the alteration in protein structure and intermolecular force before and after mutation were simulated by using a 3D protein model to initially identify suspected pathogenic mutations.Results The results of sequencing showed that there were two missense mutations in the TS BDNF gene,c.118C>T/p.H40Y,c.196G>A/p.V66M and a deletion mutation c.99_100del:p.C34Ffs 12 and c.118C>T was a rare variant that had not been reported.The bioinformatics software predicted that all three mutations were deleterious and the multispecies protein homology comparison showed that H40 and V66 were located in highly conserved regions.p.H40Y was predicted by MAESTROweb to reduce protein stability and Pymol predictions showed that the p.V66M variant altered the hydrogen bonds between amino acids,which in turn affected the stability of the protein’s spatial structure.Conclusion The BDNF missense mutations c.118C>T/p.H40Y,c.196G>A/p.V66M and deletion c.99_100del:p.C34Ffs 12 are likely to be pathogenic mutation loci in TS,which will help to expand the mutational spectrum of BDNF and provide new targets for the treatment.

关 键 词：图雷特综合征 BDNF基因 全外显子组测序 突变 

分 类 号：R749.94[医药卫生—神经病学与精神病学]