Epigenomic regulation of oxidative stress-producing innate immunity in neuroinflammation

项目总结/摘要 氧化应激是先天免疫诱导的神经退行性疾病的核心部分 包括多发性硬化症（MS）。然而，调节氧化应激基因回路的分子机制 促进神经毒性免疫反应的作用仍然没有得到很好的表征。新出现的证据支持 表观基因组在MS中紧密调节免疫细胞基因活性。然而，表观基因组的景观和功能 在促氧化剂中，MS中神经毒性中枢神经系统（CNS）先天免疫细胞仍然未知。因此，在本发明中， 能够选择性抑制免疫驱动的神经变性的药物的发现受到阻碍， 缺乏对CNS先天免疫细胞神经毒性功能的分子理解。这个项目的最终目标 该项目的目的是确定促氧化免疫细胞的调控格局，并确定翻译机制， 将表观遗传畸变转化为先天免疫驱动的神经退行性变以设计新的治疗干预 我们的初步数据发现，神经毒性小胶质细胞和外周血淋巴细胞的分子会聚， 在MS模型中，巨噬细胞与核心氧化应激基因签名的结合。通过应用创新的实验 设计与尖端的方法，这项建议的目的是定义表观遗传和转录组成部分， 氧化应激产生的先天免疫细胞在神经炎症的MS小鼠模型中通过无偏 开放染色质景观（Aim 1）和组蛋白修饰（Aim 2）的分析。这些分子 表征将确定关键的MS相关调控元件，其将在体外进行功能验证， 在体内用CRISPR干扰测定（Aim 3）。该项目将提供一个基础的表观基因组学的前景， 在神经炎症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者- 介导的氧化损伤的MS和相关条件。全面的培训计划将使PI能够 实现他的职业目标，推出一个成功的独立研究实验室，致力于研究 表观基因组机制有助于MS中的免疫功能障碍，用于靶向治疗。MOSAIC 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劳动力的多样性。作为卡特琳娜医生的学员 Akassoglou的实验室是MS发病机制的神经血管和免疫机制的领导者， Gladstone研究所和加州大学旧金山弗朗西斯科的学术环境，PI将 在K99阶段获得功能表观基因组学和CRISPR基因组工程的新培训。pi的 由具备相关专业知识的教员组成的咨询委员会将加强培训和职业发展; 以及PI参与教学活动，如表观基因组学课程和研讨会，将共同 允许PI完成该项目并整合这些方法，为 MS和表观基因组学在未来的独立研究领域。

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.