基于高通量全基因组测序技术探寻人颅内动脉瘤新易感基因的研究

Ruptured intracranial aneurysm (IA) can cause high morbidity and mortality, its formation mechanism is closely related with genetics. IA related predisposing genes are the research focus in IA genetics study, which have found that some susceptibility genes are associated with IA occurrence. Whole genome sequencing, the new high-throughput sequencing technology, can deeply excavate genetic variation at DNA level, to provide the most comprehensive genetic information for IA research. Our group has found that there are missense mutations on MAP2K3 and KCNJ18 gene located in chromosome 17, frameshift mutations on CREB3L1 gene located in chromosome 11 in familial IA patients, laying the groundwork for exploring new IA predisposing genes. On this basis, the extended FIA samples for whole genome sequencing and bioinformatics analysis; Verification genetic variation with the technology of SNP Typing in extended sporadic IA samples, further to determine the new brain aneurysm related susceptibility gene expression; detection of related susceptibility gene in intracranial aneurysm wall tissue to verify its correlation with the occurrence of IA. Our woke could screen out the new IA related predisposing genes, providing important information for the study of IA pathogenesis and genetic mechanism, providing new evidence and new targets for clinical prevention, diagnosis and treatment of IA.

颅内动脉瘤(intracranial aneurysm，IA)破裂出血可导致很高的病残率和病死率，其生成机制与遗传学密切相关。IA相关易感基因是目前IA遗传学研究热点，已发现若干与IA相关的易感基因。最新的高通量全基因组测序技术能全面挖掘 DNA 水平的遗传变异，为IA研究提供最全面的遗传信息。本组前期在家族性IA病人群中发现位于17号染色体上MAP2K3 和KCNJ18基因存在错义突变、位于11号染色体上CREB3L1基因存在移码缺失等，为探寻新的IA易感基因打下了基础。本课题将在此基础上扩大家族性IA样本进行全基因组测序和生物信息学分析；在散发IA中利用SNP分型等技术，进一步确定新的脑动脉瘤相关易感基因；检测脑动脉瘤壁组织相关易感基因的表达，验证其与IA发生的相关性。从而筛选出新的IA易感基因，为研究IA发病及遗传机制提供重要信息，为临床IA预防、诊断和治疗提供新依据和新靶点。

颅内动脉瘤破裂出血可导致很高的致残率和致死率，目前研究认为遗传因素和环境因素在颅内动脉瘤的发生发展中起到重要作用。课题组在本国自然项目（81571144）的支持下通过对颅内动脉瘤患进行二代测序，并使用先进的生物信息学方法筛选得到IA新的易感基因ARHGEF17。ARHGEF17基因与颅内动脉瘤相关的变异位点均在ARHGEF17蛋白的重要结构域，通过斑马鱼arhgef17基因敲低模型，发现arhgef17敲低可以引起斑马鱼幼鱼颅内出血，提示ARHGEF17的功能缺失可引起颅内血管损伤，结果证实ARHGEF17是IA的易感基因。同时我们在中国IA人群中验证了NOTCH3、SOX17、CDKN2A/B基因与IA的发生发症存在显著相关。项目开展期间项目组成员共发表SCI收录文章3篇，申请人以第一作者发表文章1篇，通讯作者发表文章2篇，合计影响因子13.374分，中文文章3篇，博士学位论文2篇，培养了博士研究生两名，硕士研究生6名，此外我们建立了中国多中心脑动脉瘤数据库（CMAD），联合诸多医院收集IA患者临床资料，通过大数据反映我国IA诊疗的真实情况。我们通过发现新的IA易感基因，成果丰富了IA遗传领域的基础研究，促进了IA筛查和预防工作的开展，为IA的诊断和治疗提供了新的思路。