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

项目总结 虽然存在许多转基因工具和方法来实现对 健康大脑中的许多细胞类型，旨在靶向和研究仅存在于大脑中的细胞的工具。 缺乏松弛或受损的大脑。几乎所有的神经退行性疾病，脑损伤都有 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， 它们会变成“反应性”。星形胶质细胞通常为神经元提供关键的支持，只有在 反应性星形胶质细胞(RAS)与脑部疾病和炎症一个长期存在的问题 目前尚不清楚RAS是否有助于或帮助减缓疾病的进展。这项申请的目的是- 阳离子将提供一种新的组合转基因策略和工具包，专门针对DIS中的RAS。 放松点。这个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。 在脑部疾病和炎症的发展过程中，压力只存在于RAS中。脑部感染 通过这种寄生虫，弓形虫将成为由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，以选择性地 仅在脑部疾病的RAS中操纵基因表达。在目标2中，我们将使用新方法 在不同阶段选择性地将RAS消融、阻止或重新编程为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以下方面的新信息 脑部炎症早期与持续阶段的反应性星形胶质细胞。在目标3中，我们将表演 “翻译组”分析以确定脑炎症期间反应性星形胶质细胞中独特改变的基因。 为今后的研究提供了新的靶点。我们的创新方法将使我们能够- 以前所未有的信噪比抑制基因表达的诱导和减少 接近了。拟议研究的基本原理是，提高对细胞的了解-- 脑部疾病的LAR和分子机制将为发展更多 有效的治疗方法。我们预计这项研究将是变革性的，因为我们将向 研究界研究反应性细胞类型在任何疾病中的作用的强大新策略 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.