SATB2基因缺失/突变与新生儿原因不明的神经发育障碍之间的关系及分子机制研究

项目名称： SATB2基因缺失/突变与新生儿原因不明的神经发育障碍之间的关系及分子机制研究

项目编号： No.81471483

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： 王慧君

作者单位： 复旦大学

项目金额： 66万元

中文摘要： 新生儿神经发育障碍发生率高，危害大，在排除了常见染色体异常后，很多病例难以明确诊断。SATB2是一个高度保守的转录因子，在大脑发育中占有重要地位。虽然在胼胝体发育不良、智力障碍患者中已发现SATB2基因的缺失/突变，但缺乏深入研究。我们在前期基础上，研究SATB2基因突变对大脑发育的影响。本课题中，拟通过详细的序列分析，检测SATB2基因在大样本原因未明的脑发育异常新生儿中的突变/缺失情况，并对发现的新的突变进行体外细胞株研究和体内孕鼠大脑胼胝体质粒注射和电转，分析突变体对Satb2的表达、定位和对下游分子的影响。通过与中科院神经所合作，采用CRISPR/CAS9定点基因敲除技术，建立Satb2基因R239X等突变体的小鼠模型，分析不同突变位点对小鼠胚胎大脑发育的影响并进行分子机制研究和干预探讨。通过本研究，可探索部分临床新生儿脑发育异常的原因，为临床早期干预和遗传咨询提供理论依据。

中文关键词： SATB2；新生儿；神经发育；出生缺陷；基因突变

英文摘要： Neurodevelopmental disorder is a great harmful factor threatening newborn health. The difficulty is hard to give a clear diagnosis when exclusing of common chromosomal abnormalities.SATB2 is a highly conserved transcription factor that plays an important role in brain development. Although several mutations of SATB2 have been found in clinical corpus callosum agenesis, mental retardation and other cases, in-depth study is very limited. On the basis of our previous research works, we intended to study the role of Satb2 gene mutations on the brain development. In this study, the deletion/mutations situation of SATB2 in unknown cause of abnormal brain development newborns will be detected in cohort of cases using deep next generation sequencing technology. New mutations of unknown function found in patients will be studied in vitro using HEK293 cell lines and in vivo studies using in utero electroporation, to analysis of the expression, protein location, and the effection on it's related molecular of SATB2 mutants. Meanwhile, by the cooperation with Shanghai Institutes for Biological Sciences of Chinese Academy of Sciences，we will create Satb2 mutants by means of CRISPR/CAS9 target gene knockout technology. Then, using cell biology, genetics,and behavior analysis, to explore changes caused by Satb2 gene mutations in mice brain development, the mechanisms, and possible intervening methods. The purpose of this study is to clarify Satb2 mutant effects on mouse brain development and mechanisms involved, to explore at least part of the reasons of the clinical neonatal neurodevelopmental abnormalities. This study can provide a theoretical basis for early intervention and give evidence of clinical and genetic counseling.

英文关键词： SATB2;Newborn;Neuro-development;Birth defect;Gene mutation