Functional epigenomic dissection of genetic generalized epilepsies

遗传性全面性癫痫的功能表观基因组解剖

• 批准号: 394773888
• 负责人: Professor Dr. Albert Becker
• 金额: --
• 依托单位: Institut für Neuropathologie
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/394773888?language=en
• 关键词: Functional; epigenomic; dissection; genetic; generalized

Genetic generalized epilepsies (GGEs) represent the most common form of genetically determined epilepsies. Up to date, the genetic basis of common GGE syndromes remains largely unsolved due to their prominent polygenic predisposition and extensive genetic heterogeneity. Heritability analyses implicate that at least 35% of GGE susceptibility is determined by single nucleotide polymorphisms (SNPs). Genome-wide association studies (GWASs) have identified only few GGE-risk loci mainly in non-coding genomic regions, suggesting cis-acting regulatory effects of the causal SNPs on gene expression. This project aims to dissect regulatory SNPs (rSNPs) conferring risk of GGE by epigenomic profiling of candidate SNPs derived from GWAS risk loci of GGE and candidate genes implicated in epileptogenesis. Candidate rSNPs will be prioritized in silico by their genomic intersection with binding sites of transcription factors, miRNAs and lncRNAs located within brain- and cell type-specific epigenetic elements regulating gene expression. Functional validation of the most promising candidate rSNPs will evaluate the cis-regulatory quantitative effects of rSNPs on gene expression (expression quantitative trait locus, eQTL) and CpG methylation (methylation QTL, meQTL) in human hippocampal biopsies, and in vitro gene expression analyses in primary neurons using luciferase reporter assays. Subsequently, intraventricular in utero electroporation (IUE)-mediated transgenic mouse reporter assays will explore the allelic effects of gene promoter rSNPs by in vivo molecular imaging. Initially, functional validation methods will be established for candidate rSNPs predicted to affect the promoter activity of two known epilepsy genes (CaV3.2/CACNA1H, PIGP). Together, epigenomic profiling of GGE-associated rSNPs will gain critical insights into temporospatial neurobiological processes of epileptogenesis. Predictive modeling of key pathways of epileptogenesis and delineation of individual epigenomic risk profiles of GGE may be of relevance towards precision medicine.

遗传性全身性癫痫（GGEs）是最常见的遗传性癫痫。迄今为止，常见GGE综合征的遗传基础仍然在很大程度上没有解决，由于其突出的多基因易感性和广泛的遗传异质性。遗传力分析表明，至少35%的GGE易感性是由单核苷酸多态性（SNP）决定的。全基因组关联研究（GWASs）已经确定了主要在非编码基因组区域的少数GGE风险位点，这表明因果SNP对基因表达的顺式调节作用。本项目旨在通过对GGE的GWAS风险位点的候选SNP和与癫痫发生有关的候选基因进行表观基因组分析，剖析赋予GGE风险的调节性SNP（rSNPs）。候选rSNP将通过其与位于调节基因表达的脑和细胞类型特异性表观遗传元件内的转录因子、miRNA和lncRNA的结合位点的基因组交叉在计算机上优先化。最有希望的候选rSNP的功能验证将评估rSNP对人类海马活检中基因表达（表达数量性状位点，eQTL）和CpG甲基化（甲基化QTL，meQTL）的顺式调控定量效应，以及使用荧光素酶报告基因测定法在原代神经元中进行体外基因表达分析。随后，脑室内子宫内电穿孔（IUE）介导的转基因小鼠报告基因检测将探讨基因启动子rSNPs的等位基因效应的体内分子成像。最初，将建立预测影响两个已知癫痫基因（CaV3.2/CACNA 1H，PIGP）启动子活性的候选rSNP的功能验证方法。总之，表观基因组分析的GGE相关的rSNPs将获得重要的见解癫痫发生的时空神经生物学过程。癫痫发生的关键途径的预测建模和GGE的个体表观基因组风险特征的描绘可能与精准医学有关。