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

项目摘要 虽然存在许多转基因工具和方法来使得能够操纵基因表达，但在一些实施方案中， 健康大脑中的许多细胞类型，旨在靶向和研究仅存在于疾病中的细胞的工具， 缺乏放松或受损的大脑。几乎所有的神经退行性疾病都有， 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， 其变成“反应性的”。星形胶质细胞通常为神经元提供关键支持， 反应性星形胶质细胞（RA）在脑疾病和炎症中的作用。一个长期存在的问题， 目前尚不清楚RA是否有助于或有助于缓解疾病进展。本申请的目的是-- 目的是提供一种新的组合转基因策略和工具包，以特异性靶向RA， 放松。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）基因表达。 在脑部疾病和炎症进展的任何时候，RA中的表达都是有限的。脑部感染 寄生虫弓形虫将作为一个模型的脑部炎症源于感染。 提出了三个目标：在目标1中，我们将首先表征Cre转基因策略，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 在不同阶段选择性消融、预防RA或将RA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。我们的工作将提供新的信息， 反应性星形胶质细胞在大脑炎症的早期与持续阶段。在目标3中，我们将 “translatome”分析，以确定在脑炎症过程中反应性星形胶质细胞中独特改变的基因， 这是第一次，为未来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都具有前所未有的信噪比， 他走近了。这项研究的基本原理是，提高对细胞的理解， 大脑疾病的更大和分子机制将为更多的发展提供新的见解。 有效的治疗。我们预计，这项研究将是变革性的，因为我们将介绍给 研究社区一个强大的新策略，以调查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.