The role of histone demethylase KDM5B in ethanol-induced microglial activation

组蛋白去甲基化酶 KDM5B 在乙醇诱导的小胶质细胞激活中的作用

• 批准号: 9617532
• 负责人: Stanley M Stevens
• 金额: $ 37.97万
• 依托单位: ALBANY COLLEGE OF PHARMACY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-10 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9617532
• 关键词: role; histone; demethylase; KDM5B; ethanol

Microglia serve as resident immune cells in the brain and are considered key contributors to neuroinflammation, a process that has been implicated in the negative, long-term effects of alcohol on the central nervous system (CNS). The molecular mechanisms underlying activation and related phenotypic transitions of microglia during ethanol exposure are unclear. Based on a recent proteomic analysis of ethanol- treated microglia, we can demonstrate that a significant portion of the ethanol-induced proteome response in microglia could be attributed to changes in the activity of KDM5B, a histone demethylase that catalyzes the removal of tri-methylation on Lys 4 of Histone H3 (H3K4me3). Moreover, we have strong preliminary data that show ethanol induces histone methylation changes both in vitro and in vivo and that experimental modulation of KDM5B activity affects the pro-inflammatory response of microglia. Therefore, we hypothesize that methylation is an important epigenetic modification that influences ethanol-induced activation of microglia resulting in unique functional outcomes that have immediate and possible transgenerational effects on the brain. In order to test our hypothesis, we plan to 1) characterize ethanol-induced changes in the microglial histone methylation code mediated by KDM5B in both primary pure microglia cultures and neuron-microglia co- cultures, 2) determine ethanol dose- and time-dependent role of KDM5B and related histone methylation changes on ethanol-induced microglial activation in vivo and 3) characterize the functional outcome of ethanol- induced epigenetic inheritance of KDM5B-mediated methylation in microglia. The proposed studies incorporate novel approaches such as activity-based protein profiling, methylation-specific quantitative mass spectrometry and ChIP-Seq in order to carry out these aims. This project will be the first ever comprehensive global-scale analysis of methylation and its regulators, which could determine, at least partly, the epigenetic code related to microglial activation phenotype after chronic ethanol exposure in the brain. Characterization of these ethanol- responsive pathways in microglia could also lead to further insight into the development of novel epigenetic therapies for the treatment of CNS dysfunction resulting from alcohol abuse.

小胶质细胞是大脑中的常驻免疫细胞，被认为是免疫细胞的关键贡献者 神经炎症，这一过程与酒精对神经系统的负面、长期影响有关 中枢神经系统（CNS）。激活和相关表型的分子机制 乙醇暴露期间小胶质细胞的转变尚不清楚。基于最近对乙醇的蛋白质组学分析 经过处理的小胶质细胞，我们可以证明乙醇诱导的蛋白质组反应的很大一部分 小胶质细胞的变化可能归因于 KDM5B 活性的变化，KDM5B 是一种组蛋白去甲基化酶，可催化 去除组蛋白 H3 (H3K4me3) 的 Lys 4 上的三甲基化。此外，我们有强有力的初步数据表明 显示乙醇在体外和体内诱导组蛋白甲基化变化，并且实验调节 KDM5B 活性的影响影响小胶质细胞的促炎反应。因此，我们假设 甲基化是一种重要的表观遗传修饰，影响乙醇诱导的小胶质细胞激活 产生独特的功能结果，对身体产生直接和可能的跨代影响 脑。为了检验我们的假设，我们计划 1）表征乙醇诱导的小胶质细胞变化 原代纯小胶质细胞培养物和神经元-小胶质细胞共培养物中由 KDM5B 介导的组蛋白甲基化代码 培养物，2) 确定 KDM5B 和相关组蛋白甲基化的乙醇剂量和时间依赖性作用 体内乙醇诱导的小胶质细胞激活的变化，以及3）表征乙醇的功能结果- 诱导小胶质细胞中 KDM5B 介导的甲基化的表观遗传。拟议的研究包括 新方法，例如基于活性的蛋白质分析、甲基化特异性定量质谱分析 和 ChIP-Seq 以实现这些目标。该项目将是第一个全球范围内的综合性项目 甲基化及其调节因子的分析，这可以至少部分地确定与以下相关的表观遗传密码 大脑中慢性乙醇暴露后的小胶质细胞活化表型。这些乙醇的表征- 小胶质细胞中的反应途径也可能导致进一步深入了解新型表观遗传的发展 用于治疗因酗酒引起的中枢神经系统功能障碍的疗法。