Exploring the role of reactive astrocytes in brain inflammation using a novel combinatorial strategy

项目概要 虽然存在许多转基因工具和方法来操纵基因表达 健康大脑中存在多种细胞类型，旨在针对和研究仅存在于大脑中的细胞的工具 缺乏缓解或受损的大脑。几乎所有神经退行性疾病、脑损伤都很常见 感染是一种神经炎症和免疫反应，其特征是星形胶质细胞的变化， 变得“反应性”。星形胶质细胞通常为神经元提供关键支持，并且只会变成 反应性星形胶质细胞（RA）在脑部疾病和炎症中的作用。一个长期存在的问题一直存在 目前尚不清楚 RA 是否有助于或有助于缓解疾病进展。本申请的目的 阳离子是提供一个新的组合转基因策略和工具包，专门针对 RAs 轻松。该工具包将使研究人员能够选择性地改变（消除、增加或减少）基因前 仅在脑部疾病和炎症进展过程中的任何时刻，RA 中才会出现抑制。脑部感染 弓形虫寄生虫将作为感染引起的脑部炎症的模型。 提出了三个目标： 在目标 1 中，我们将首先描述 Cre 转基因策略的特征，以选择性地 仅在脑部疾病的 RA 中操纵基因表达。在目标 2 中，我们将使用新方法 选择性地消融、预防 RA 或将 RA 重新编程回不同阶段的非反应性星形胶质细胞 在急性和慢性炎症过程中。我们的工作将提供有关角色的新信息 脑炎症早期与持续阶段的反应性星形胶质细胞。在目标 3 中，我们将执行 “翻译组”分析可识别脑炎症期间反应性星形胶质细胞中独特改变的基因 首次实现这一目标，为今后的研究提供了新的目标。我们的创新方法将允许检测 基因表达的诱导和减少具有前所未有的信噪比 列出方法。拟议研究的基本原理是提高对细胞的理解 脑部疾病的分子机制将为更多疾病的发展提供新的见解 有效的治疗。我们预计这项研究将具有变革性，因为我们将介绍 研究界提出了一种强大的新策略来研究反应性细胞类型在任何疾病中的作用 或神经系统紊乱。

PROJECT SUMMARY While numerous transgenic tools and approaches exist to enable manipulation of gene expression in many cell types in the healthy brain, tools designed to target and study cells present only in the dis- eased or damaged brain are lacking. Common to virtually all neurodegenerative diseases, brain injuries and infections is a neuroinflammatory and immune response characterized by changes in astrocytes, which become “reactive”. Astrocytes ordinarily provide critical support for neurons and only turn into reactive astrocytes (RAs) in brain disease and inflammation. A longstanding issue which has remained unknown is whether RAs contribute to or help alleviate disease progression. The objective of this appli- cation is to deliver a new combinatorial transgenic strategy and toolkit to specifically target RAs in dis- ease. This toolkit will enable researchers to selectively alter (eliminate, increase, or decrease) gene ex- pression only in RAs at any point in the progression of brain disease and inflammation. Brain infection by the parasite Toxoplasma gondii will serve as a model of brain inflammation stemming from infection. Three aims are proposed: In Aim 1, we will first characterize the Cre transgenic strategy to selectively manipulate gene expression only in RAs in brain disease. In Aim 2, we will then use the new approach to selectively ablate, prevent, or reprogram RAs back into non-reactive astrocytes at various stages during the acute and chronic inflammatory process. Our work will provide new information on the role of reactive astrocytes in the early vs. sustained stages of brain inflammation. In Aim 3, we will perform “translatome” analysis to identify genes uniquely altered in reactive astrocytes during brain inflamma- tion for the first time, providing novel targets for future study. Our innovative approach will allow detec- tion of both inductions and reductions in gene expression with unprecedented signal-to-noise over ex- isting approaches. The rationale for the proposed research is that improving understanding of the cellu- lar and molecular mechanisms of brain disease will provide novel insights into the development of more effective treatments. We anticipate that this research will be transformative, as we will introduce to the research community a powerful new strategy to investigate the role of reactive cell types in any disease or disorder of the nervous system.