PICK1基因突变致癫痫的发病机制及其分子诊断方法研究

项目名称： PICK1基因突变致癫痫的发病机制及其分子诊断方法研究

项目编号： No.81501824

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 王震

作者单位： 杭州医学院

项目金额： 18万元

中文摘要： 癫痫是神经元同步放电功能异常的疾病，我国的癫痫患者高达900多万。有研究表明失神性癫痫与PICK1对mGluR7a的调控密切相关，但具体发病机制不清楚，也尚无合适的分子诊断方法。我们的前期研究发现癫痫患儿存在PICK1基因嵌合突变，本课题研究中我们拟进一步增加研究例数，对新发现的和已报道的PICK1突变进行致癫痫发病机制研究。研究中，在构建不同的PICK1突变基因，ICA69和mGluR7等克隆后，分别利用培养的神经元，HEK293T和PICK1 KO小鼠脑组织切片进行共定位，膜运输，神经突触可塑性等机制研究。对癫痫发病机制深入理解的基础上，我们希望建立简便、灵敏的分子诊断方法即利用q-PCR或d-PCR方法检测癫痫患儿和其父母的主要PICK1及相关基因的突变位点。该方法一旦建立，也较易推广到其它癫痫基因或疾病的分子诊断，从而为癫痫性患儿的病因诊断及其父母、家属的优生优育提供指导和依据。

中文关键词： PKC结合蛋白；癫痫；分子诊断；胰岛细胞自身抗原69KD蛋白；代谢性谷氨酸受体7a

英文摘要： Epilepsy is a disease of synchronized discharge dysfunction of neurons. There are more than 900 million of epilepsy patients in China. It has been proved that absence epilepsy is closely related with the regulation of mGluR7a by PICK1. However, the exact pathogenesis is still unclear, and currently there is no proper molecular diagnostic method. Our previous study found chimeric gene mutations of PICK1 in epilepsy patients. In this project, we intend to increase the number of patients studied, and investigate the pathogenesis of epilepsy caused by PICK1 mutations, including the clinical findings and those reported in articles. The plasmids of the mutants of PICK1, ICA69 and mGluR7 were constructed and applied to the cultured neurons, HEK293T and brain tissue sections of PICK1 KO mouse for exploring the mechanism of co-localization, membrane traffic and synaptic plasticity. Based on the further understanding of the pathogenesis of epilepsy, it is possible to establish simple and sensitive molecular diagnostic methods. That is, q-PCR or d-PCR methods will be applied to detect mutations of PICK1 and related genes in epilepsy children as well as their parents. The method, once established, might be also useful for the study of other epilepsy genes or for the molecular diagnostic detection of other diseases. Thus it will be a breakthrough for the etiological diagnosis of epilepsy in children and prenatal guidance will be provided for their parents and relatives.

英文关键词： PICK1;epilepsy;Molecular Diagnosis;ICA69;mGluR7a