Exosomes: A Novel Contributor of Microglia-Mediated Neuroinflammation and Neurodegeneration.

外泌体：小胶质细胞介导的神经炎症和神经变性的新贡献者。

• 批准号: 10653688
• 负责人: Juliet Victoria Santiago
• 金额: $ 4.77万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10653688
• 关键词: Adopted; Alzheimer' s Disease; Alzheimer' s disease pathology; Amyloid beta-Protein; Anti-Inflammatory Agents; Astrocytes; Biological; Biotin; Biotinylation; Brain; Cells; Cognition; Cytoplasm; Dementia; Deterioration; Disease; Disease associated microglia; Goals; Immune; In Vitro; Inflammatory; Knowledge; Label; Ligase; LoxP-flanked allele; Mass Spectrum Analysis; Measures; Mediating; Memory; Messenger RNA; MicroRNAs; Microglia; Molecular; Mutate; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurons; Pathogenesis; Pathogenicity; Pathologic; Pathology; Persons; Phenotype; Play; Proteins; Proteome; Proteomics; Research; Rest; Role; Synapses; Testing; Therapeutic; Wild Type Mouse; Work; abeta accumulation; abeta oligomer; beta amyloid pathology; brain cell; cognitive function; cytokine; cytotoxicity; effective therapy; exosome; extracellular vesicles; in vitro Model; in vivo; in vivo Model; inhibitor; insight; intercellular communication; macromolecule; mouse model; neuroinflammation; neurotoxic; neurotoxicity; new therapeutic target; novel; polarized cell; protein aggregation; protein biomarkers; proteomic signature; response; tau Proteins; therapeutic target; transcriptomics

PROJECT ABSTRACT: Alzheimer’s disease (AD) is a devastating and rapidly rising neurodegenerative disorder, afflicting nearly 44 million people worldwide. AD pathology includes progressive protein aggregation of amyloid-beta (Aβ) and tau. A key pathological component of AD is neuroinflammation which is mediated by immune cells of the brain called microglia. Although microglia-mediated neuroinflammation has emerged as a causal disease mechanism, there are still critical gaps in our understanding of what microglial protein changes occur in AD and how microglia can perpetuate AD pathology. Exosome release has emerged as a mechanism of microglia-mediated neuroinflammation and neurodegeneration. Given the critical role of exosomes in the transfer of macromolecules between cells to facilitate intercellular communication, it is possible that microglia-derived exosomes transfer pathogenic cargo which could perpetuate AD. Furthermore, microglia can adopt different states, and therefore, exosomes derived from these microglia states are likely to have different compositions and distinct effects. However, the role of microglia-derived exosomes in neuroinflammation and AD pathology remains poorly investigated. To this end, there is a pressing need to define microglial protein level changes in AD and define the molecular composition and state-specific effects of microglia-derived exosomes in AD pathology. The overall goal of my F31 proposal is to understand how microglia and microglia-derived exosomes perpetuate AD pathology. I will test the hypothesis that the protein profiles of microglia and microglia-derived exosomes impact AD pathology and neuroinflammation. In Aim 1, I will determine the effect of microglia state on exosome composition and exosome-mediated responses in-vitro. I will polarize microglia to either a homeostatic, anti- inflammatory, pro-inflammatory, or “AD-like” state using a combination of different cytokines and Aβ oligomers. I will then measure the ability of exosomes released by these microglia to induce pro-inflammatory changes in resting glial cells and cytotoxicity in neurons. Using mass spectrometry, I will also define differences in the protein profiles of exosomes derived from different microglial states. In Aim 2, I will define changes in microglia-derived exosomal proteome that are associated with AD pathology in-vivo. To accomplish this, I will employ a mouse model of Aβ pathology (5XFAD), a novel in-vivo protein labeling strategy which uses a mutated biotin ligase (TurboID) to biotinylate proteins in microglia and microglia-derived exosomes, and quantitative proteomics using mass spectrometry. The results from these studies will advance our understanding of neuroinflammatory mechanisms of AD pathogenesis and contribute to the discovery of therapeutic targets for AD.

项目摘要： 阿尔茨海默病(AD)是一种破坏性的、迅速上升的神经退行性疾病，几乎 全球有4400万人。AD病理包括进行性淀粉样β蛋白聚集(Aβ)和 陶先生。阿尔茨海默病的一个关键病理成分是由大脑免疫细胞介导的神经炎症 叫做小胶质细胞。尽管小胶质细胞介导的神经炎症已经成为一种致病机制， 我们对阿尔茨海默病的小胶质细胞蛋白的改变以及小胶质细胞如何改变的理解仍然存在严重的差距。 会使AD的病理永久化。外切体释放已成为小胶质细胞介导的一种机制 神经炎症和神经变性。鉴于外体在大分子转移中的关键作用 为了促进细胞间的交流，小胶质细胞衍生的外切体转移是可能的 可能使AD永久化的致病货物。此外，小胶质细胞可以采取不同的状态，因此， 来自这些小胶质细胞状态的外切体可能具有不同的组成和不同的作用。 然而，小胶质细胞来源的外切体在神经炎症和AD病理中的作用仍然很少。 调查过了。为此，迫切需要确定AD中小胶质细胞蛋白水平的变化，并确定 小胶质细胞来源的外切体在AD病理中的分子组成和状态特异性效应。整体而言 我的F31提案的目标是了解小胶质细胞和小胶质细胞衍生的外切体如何使AD永久化 病理学。我将检验这样一个假设，即小胶质细胞和小胶质细胞衍生的外切体的蛋白质谱影响 AD病理和神经炎症。在目标1中，我将确定小胶质细胞状态对外切体的影响 组成和外切体介导体的体外反应。我会把小胶质细胞极化成一种动态平衡，抗- 炎症性、促炎性或类似AD的状态，使用不同的细胞因子和Aβ寡聚体的组合。 然后我将测量这些小胶质细胞释放的外切体在体内诱导促炎变化的能力 静息的神经胶质细胞和神经元的细胞毒性。利用质谱学，我还将确定蛋白质之间的差异 来自不同小胶质细胞状态的外切体谱。在目标2中，我将定义小胶质细胞来源的变化 与体内AD病理相关的外体蛋白质组。为了实现这一点，我将使用一只鼠标 β病理模型(5XFAD)--一种使用突变的生物素连接酶的体内蛋白质标记新策略 (TurboID)对小胶质细胞和小胶质细胞衍生的外切体中的生物素化蛋白，以及使用 质谱学。这些研究的结果将促进我们对神经炎性疾病的理解 研究阿尔茨海默病的发病机制，有助于发现AD的治疗靶点。