Epigenomic regulation of oxidative stress-producing innate immunity in neuroinflammation

项目摘要/摘要 氧化应激是先天免疫诱导的神经性疾病中神经变性的核心部分 包括多发性硬化症(MS)。然而，调控氧化应激基因回路的分子机制 促进神经毒性免疫反应的特点仍不明确。新出现的证据支持 表观基因组在密切调控MS免疫细胞基因活性中的作用 在促氧化剂中，MS中的神经毒性中枢神经系统(CNS)天然免疫细胞尚不清楚。因此， 能够选择性抑制免疫驱动的神经变性的药物的发现受到了以下因素的阻碍 对中枢神经系统天然免疫细胞的神经毒性功能缺乏分子认识。这样做的最终目的是 该项目是定义促氧化免疫细胞的调控格局，并确定翻译的机制 先天免疫驱动的神经变性的表观遗传学异常用于设计新的治疗干预措施 对于MS，我们的初步数据发现了神经毒性小胶质细胞和外周血细胞的分子聚集 巨噬细胞在MS模型中的核心氧化应激基因特征。通过应用一种创新的实验 设计采用尖端方法，这项建议旨在定义表观遗传和转录成分 氧化应激在多发性硬化症小鼠神经炎症模型中产生的天然免疫细胞 描述开放染色质景观(目标1)和组蛋白修饰(目标2)。这些分子 特征将确定关键的MS相关调节元件，这些元件将在体外和 体内CRISPR干扰试验(AIM 3)。这个项目将提供一个基本的表观基因组学前景 神经炎性疾病中控制促氧化剂、神经毒性免疫反应的分子回路，以及 研究结果可能会揭示开发先天免疫新疗法的候选药物- MS及相关条件下介导的氧化损伤。全面的培训计划将使PI能够 实现了他的职业目标，成立了一个成功的独立研究实验室，致力于 多发性硬化症免疫功能障碍的表观基因组学机制。马赛克UE5 指导、领导力和多样性培训将促进他向独立的过渡，并使PI能够 在R00阶段及以后加强生物医学工作人员的多样性。作为《卡特琳娜医生》的学员 在多发性硬化症发病机制的神经血管和免疫机制方面处于领先地位的Akassoglou实验室 Gladstone学院和加州大学旧金山分校受人尊敬的学术环境，PI将 在K99阶段获得功能表观基因组学和CRISPR基因组工程方面的新培训。少年派的 培训和职业发展将通过一个由具有相关专业知识的教员组成的咨询委员会予以支持； 而PI参与的教学活动，如表观基因组学的课程作业和研讨会，将共同 允许PI完成这一项目，并整合这些方法，为 未来独立研究中的MS和表观基因组学领域。

PROJECT SUMMARY / ABSTRACT Oxidative stress is a central part of innate immune-induced neurodegeneration in neurological disorders including multiple sclerosis (MS). However, the molecular mechanisms regulating oxidative stress gene circuits to promote neurotoxic immune responses remain poorly characterized. Emerging evidence supports a role for the epigenome in tightly regulating immune cell gene activity in MS. Yet, the epigenomic landscape and function in prooxidant, neurotoxic central nervous system (CNS) innate immune cells in MS remains unknown. Thus, discovery of drugs capable of selectively suppressing immune-driven neurodegeneration has been hindered by lack of molecular understanding of neurotoxic functions of CNS innate immune cells. The ultimate goal of this project is to define the regulatory landscape of prooxidant immune cells and identify mechanisms that translate epigenetic aberrations into innate-immune driven neurodegeneration for devising novel therapeutic interventions for MS. Our preliminary data discovered a molecular convergence of neurotoxic microglia and peripheral macrophages to a core oxidative stress gene signature in MS model. By applying an innovative experimental design with cutting-edge methods, this proposal aims to define the epigenetic and transcriptional components of oxidative stress-producing innate immune cells in a mouse model of neuroinflammation for MS through unbiased profiling of the open chromatin landscapes (Aim 1) and histone modifications (Aim 2). These molecular characterizations will identify key MS-related regulatory elements that will be functionally validated in vitro and in vivo with CRISPR interference assays (Aim 3). This project will provide a foundational epigenomic outlook on the molecular circuits governing prooxidant, neurotoxic immune responses in neuroinflammatory disease, and the research outcomes may reveal candidates for the development of new treatments for innate immune- mediated oxidative injury in MS and related conditions. The comprehensive training plan will enable the PI to achieve his career goal of launching a successful independent research laboratory dedicated to studying epigenomic mechanisms contributing to immune dysfunction in MS for targeted treatments. The MOSAIC UE5 mentoring, leadership, and diversity training will facilitate his transition to independence and enable the PI to enhance diversity in the biomedical workforce in the R00 phase and beyond. As a mentee in Dr. Katerina Akassoglou’s laboratory, a leader in neurovascular and immune mechanisms of MS pathogenesis, at the esteemed academic environment of Gladstone Institutes and University of California, San Francisco, the PI will obtain new training in functional epigenomics and CRISPR genome engineering during the K99 phase. The PI’s training and career development will be bolstered through an advisory committee of faculty with related expertise; and the PI’s participation in didactic activities such as coursework in epigenomics and seminars, will collectively allow the PI to complete this project and integrate these approaches for making meritorious contributions to the fields of MS and epigenomics in future independent research.