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

摘要(摘自R01DA048815)。虽然存在许多转基因工具和方法来操纵 在健康大脑中发现的许多细胞类型的基因表达，设计用于靶向和研究细胞的工具仅存在 在患病或受损的大脑中缺乏。几乎所有的神经退行性疾病，脑损伤都有 感染是以星形胶质细胞变化为特征的神经炎性和免疫反应。这些 细胞通常为神经元提供关键支持，但在脑部疾病和疾病中成为反应性星形胶质细胞(RAS) 发炎。一个长期存在且仍未解决的问题是RAS是否有助于或有助于缓解疾病 进步。这个应用程序的目标是向神经科学界提供一个新的组合 针对疾病中RAS的转基因策略和工具包。该工具包将使研究人员能够 选择性地改变(即，消除、增加或减少)仅在RAS中的任何点的基因表达 脑部疾病和炎症的进展。 将分析急性和慢性使用甲基苯丙胺的星形胶质细胞种群。提出了三个目标： 在目标1中，我们将描述仅在RAS中选择性地操纵基因表达的Cre转基因策略 在患病的大脑中。在目标2中，我们将使用新方法选择性地将RAS消融、防止或重新编程为 急性和慢性炎症过程中不同阶段的非RAS。我们的工作将提供新的 关于RAS在早期和持续脑部炎症中的作用的信息。在目标3中，我们将表演 “翻译组”分析以确定在脑部炎症期间反应性星形胶质细胞中唯一改变的基因 首次为今后的研究提供了新的靶点。我们的创新方法将允许检测到 与现有方法相比，以前所未有的信噪比诱导和减少基因表达。这个 提出这项研究的理由是，提高对细胞和分子机制的理解 脑部疾病的研究将为开发更有效的治疗方法提供新的见解。我们预料到 这项研究将具有变革性，因为我们将向研究界介绍一项强大的新战略，以 研究反应细胞类型在任何神经系统疾病或紊乱中的作用。

SUMMARY (from R01DA048815). While numerous transgenic tools and approaches exist for manipulation of gene expression in many cell types found in a healthy brain, tools designed to target and study cells present only in the diseased or damaged brain are lacking. Common to virtually all neurodegenerative diseases, brain injuries and infections are neuroinflammatory and immune responses characterized by changes in astrocytes. These cells ordinarily provide critical support for neurons, but become reactive astrocytes (RAs) in brain disease and inflammation. A longstanding and still unresolved issue is whether RAs contribute to or help alleviate disease progression. The objective of this application is to deliver to the neuroscience community a new combinatorial transgenic strategy and toolkit to specifically target RAs in disease. This toolkit will enable researchers to selectively alter (i.e., eliminate, increase, or decrease) gene expression only in RAs at any point in the progression of brain disease and inflammation. Astrocyte populations from acute and chronic methamphetamine use will be analyzed. Three aims are proposed: In Aim 1, we will characterize the Cre transgenic strategy to selectively manipulate gene expression only in RAs in diseased brains. In Aim 2, we will use the new approach to selectively ablate, prevent, or reprogram RAs into non-RAs at various stages of the acute and chronic inflammatory processes. Our work will provide new information on the role of RAs in the early vs. sustained stages of brain inflammation. In Aim 3, we will perform “translatome” analysis to identify the genes uniquely altered in reactive astrocytes during brain inflammation for the first time, providing novel targets for future study. Our innovative approach will allow detection of both inductions and reductions in gene expression with unprecedented signal-to-noise over existing approaches. The rationale for the proposed research is that improving understanding of the cellular and molecular mechanisms of brain disease will provide novel insights for 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.