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.

小胶质细胞作为大脑中的常驻免疫细胞， 神经炎症，这一过程与酒精对神经系统的长期负面影响有关。 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- 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显着部分， KDM 5 B是一种组蛋白去甲基化酶，它催化小胶质细胞的增殖。 去除组蛋白H3（H3 K4 me 3）的Lys 4上的三甲基化。此外，我们有强有力的初步数据， 显示乙醇在体外和体内均诱导组蛋白甲基化变化，且实验调节 KDM 5 B的活性影响小胶质细胞的促炎反应。因此，我们假设 甲基化是影响乙醇诱导的小胶质细胞活化的重要表观遗传修饰 导致独特的功能结果，对儿童的健康产生直接和可能的代际影响。 个脑袋为了验证我们的假设，我们计划1）描述乙醇诱导的小胶质细胞的变化， 在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KDM 5 B介导的组蛋白甲基化编码 2）确定KDM 5 B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活化的变化和3）表征乙醇- 诱导小胶质细胞中KDM 5 B介导的甲基化的表观遗传。拟议的研究包括 新的方法，如基于活性的蛋白质谱，甲基化特异性定量质谱 和ChIP-Seq来实现这些目标。该项目将是有史以来第一个全面的全球规模 分析甲基化及其调控因子，这可以确定，至少部分，表观遗传密码相关的 脑中慢性乙醇暴露后的小胶质细胞活化表型。这些乙醇的特性- 小胶质细胞中的反应通路也可能导致进一步了解新的表观遗传学的发展。 用于治疗由酒精滥用引起的CNS功能障碍的疗法。