Probing the Protective Role of EZH2 in Epilepsy

探讨 EZH2 在癫痫中的保护作用

• 批准号: 10398140
• 负责人: RAYMOND J DINGLEDINE
• 金额: $ 50.7万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10398140
• 关键词: Alzheimer' s Disease; Amygdaloid structure; Anticonvulsants; Antiepileptogenic; Appearance; Binding; Biochemical; Bioinformatics; Blood - brain barrier anatomy; Brain; Cell Death; Cell Nucleus; Cells; Cellular Assay; Chromatin; Chronic; Complex; Convulsants; Data; Data Set; Disease; Disease Progression; Dose; EZH2 gene; Epilepsy; Epileptogenesis; Gene Expression; Generations; Genes; Genetic Models; Genetic Transcription; Hippocampus (Brain); Histones; Immunohistochemistry; Incidence; Individual; Inflammation; Institution; Life; Measures; Messenger RNA; Methylation; Methyltransferase; Migraine; Modeling; Molecular Biology; Molecular Genetics; Mus; Nerve Degeneration; Neurons; Outcome Measure; Output; Pathologic; Patients; Persons; Pharmaceutical Preparations; Pharmacology; Polycomb; Population; Prosencephalon; Proteins; Publishing; Rattus; Recurrence; Resected; Ribosomes; Role; Seizures; Status Epilepticus; Stimulus; Temporal Lobe Epilepsy; Testing; Time; Transcript; Up-Regulation; Work; antagonist; axonal sprouting; bioinformatics tool; dravet syndrome; epigenetic silencing; experimental study; follow-up; gene repression; genetic approach; genetic corepressor; genome-wide; granule cell; inhibitor; kainate; mRNA Expression; nervous system disorder; neurogenesis; novel; post stroke; programs; protein complex; response; stoichiometry; transcription factor; transcriptome sequencing; virtual; whole genome

Project Summary/Abstract Epilepsy is the 4th most prevalent neurological disorder after stroke, Alzheimer’s and migraine with an incidence of 1 in 26 individuals. Though there are a number of anti-convulsant drugs available, there are no anti-epileptogenic drugs that mitigate the progression of the disease. Using novel bioinformatic approaches, we have identified an endogenous, protective program launched by the brain after a prolonged seizure that functions to mitigate pathological changes. Epileptogenesis is associated with a plethora of changes in the brain including alterations in plasticity, cell death, neurogenesis, inflammation and axonal sprouting. These changes occur over timescales ranging from many minutes to years, but the orchestrating mechanisms are virtually unknown. Long-term changes in gene expression that are associated with epileptogenesis imply that one or more master regulators of transcription may be coordinating the brain alterations. In order to uncover these transcriptional mechanisms, we turned to our recently published genome-wide expression datasets generated by the Epilepsy Microarray Consortium (EMC). The datasets consist of mRNA expression profiles of rat dentate granule cells assayed at various time points after Status Epilepticus (SE). Using a novel bioinformatic tool that integrates whole genome transcription factor binding data with gene expression profiles, we analyzed datasets derived from brains induced by 3 different convulsant stimuli, each in 2 independent labs, and at various time-points. This analysis projected that Polycomb target genes represent the majority of chronically altered genes during epileptogenesis. REST targets represent a second, overlapping, group of repressed genes. Polycomb is a well-known driver of life-long changes in gene expression that works by epigenetically silencing genes across the phyla. Our data shows an extremely robust induction of EZH2 protein (the catalytic methylase subunit of Polycomb) over a 20 day window post SE in neurons. Further, we find that antagonizing EZH2 shortly after SE robustly accelerates the onset of spontaneous recurrent seizures in mice, suggesting a protective rather than pathological role for EZH2. How antagonism of EZH2 later after SE remains to be determined. In this project, we will test the hypothesis that an alteration in Polycomb output is a principal modifier of epileptogenesis. We will ascertain whether EZH2 upregulation is always protective or whether its role evolves during the latent period. We will test the effect of an order-of-magnitude change in EZH2 levels on corepressor function to see whether such upregulation augments or hampers the repressive abilities of two major EZH2 containing complexes: Polycomb and REST. We anticipate that these studies will establish Polycomb as a major orchestrator of the long-term changes associated with epileptogenesis. If so, approaches that modulate Polycomb function may be of benefit to the 65 million people world-wide that live with epilepsy.

项目摘要/摘要 癫痫是中风后，阿尔茨海默氏症和偏头痛的第四最普遍的神经系统疾病 26个人中有1人的发病率。尽管有许多抗惊人药物可用，但没有 减轻疾病进展的抗癫痫药。使用新颖的生物信息学方法， 我们已经确定了一个长时间的癫痫发作后，大脑发起的一个内源性，受保护的程序 功能减轻病理变化。 癫痫生成与大脑的大量变化有关，包括可塑性的改变， 细胞死亡，神经发生，感染和轴突发芽。这些变化发生在时间尺度上 从数分钟到几年，但是精心策划的机制几乎是未知的。长期变化 与癫痫发生相关的基因表达意味着一个或多个主调节剂 转录可以协调大脑的改变。为了发现这些转录机制， 我们转向癫痫微阵列产生的最近发表的全基因组表达数据集 财团（EMC）。数据集由大鼠齿状颗粒细胞的mRNA表达分布组成。 状态癫痫病（SE）之后的各种时间点。使用一种整合整体的生物信息学工具 基因组转录因子与基因表达谱的结合数据，我们分析了从 由3种不同的抽搐刺激引起的大脑，每个刺激在2个独立实验室中，以及各个时间点。这 分析预测，polycomb靶基因代表了大多数慢性改变基因 癫痫发生。 REST目标代表第二个重叠的反射基因。 polycomb是a 众所周知的终身基因表达变化驱动力，通过表观遗传沉默的基因跨越跨的基因表达 门。我们的数据显示了EZH2蛋白的极强诱导（催化甲基酶亚基 polycomb）在神经元的SE后20天的窗户上。此外，我们发现不久之后拮抗Ezh2 SE坚强地加速了小鼠的赞助复发性癫痫发作，这表明了保护性 比EZH2的病理作用。在SE之后后，EZH2的拮抗作用如何仍有待确定。 在这个项目中，我们将检验以下假设：Polycomb输出的改变是 癫痫发生。我们将确定EZH2上调是否始终受到保护，还是它的作用演变 在潜在时期。我们将测试EZH2级别的速率变化对CorePressor的影响 功能以查看这种上调是否增加或阻碍了两个主要EZH2的反射能力 包含复合物：polycomb和Rest。我们预计这些研究将建立polycomb 与癫痫发生相关的长期变化的主要编排。如果是这样，请接近 调节PolyComb功能可能对全球6500万人有利 癫痫。