Functional Networks in Microglia and Astrocytes Regulating Neuropathology of Alzheimer's Disease

项目摘要 阿尔茨海默病（Alzheimer's disease，AD）是老年痴呆症最常见的病因，目前尚无有效的治疗方法。 治疗。选择性神经元脆弱性，淀粉样β（Aβ）斑块的出现和积聚，改变 电路功能、突触损失和退化是AD的标志。小胶质细胞和星形胶质细胞，两大 神经胶质细胞的亚型，有助于这些神经病理学变化。然而，精确的机制 控制这些过程仍然是难以捉摸的。其中一个原因可能是我们仍然缺乏关于 细胞类型特异性改变以及它们如何促进AD的发作和进展。这里我们 假设通过揭示小胶质细胞和星形胶质细胞中基因功能网络的详细变化， 这些过程是如何交叉的，我们也许能够确定它们对神经病理学变化的贡献， AD.我们将利用翻译核糖体亲和纯化结合新的生物信息学工具， 构建胶质细胞特异性功能的功能网络模型，并将其与AD定量遗传学相结合， 数据使用这个框架，我们将检测胶质细胞特异性基因最有可能与AD病理学相关， 无偏数据驱动的方式，进一步研究星形胶质细胞和 小胶质细胞将在体外检查来自该分析的相关候选基因在AD进展中的作用， 并将重点放在细胞因子受体途径，特别是白细胞介素1和集落 刺激因子1枢纽。我们将利用原代细胞培养和与星形胶质细胞和小胶质细胞共培养 分离自健康和5xFAD小鼠。此外，我们将只研究与基因表达相关的基因。 人类病理学为了实现这一点，我们将测试AD标志和/或救援表型的出现， 从AD患者分离的诱导多能干细胞（iPSC）衍生的小胶质细胞和星形胶质细胞 健康的对照。最后，最重要的候选基因将通过基因表达进行体内测试 在适当的小鼠品系中进行操作。我们将测量：i）Aβ蓄积、摄取和降解， 小胶质细胞和星形胶质细胞，ii）神经元变性，iii）突触功能，包括突触标志物，以及 功能电路，iv）斑块形成，和v）认知功能，用一系列行为测试评估。 拟议的研究将揭示每种细胞类型的分子概况，并检查它们的相互作用。生成 数据集和生物信息学工具将通过网络界面与公众共享。这些数据将提供 新的见解的外观AD的标志和阐明疾病的基本机制。 研究这些变化，特别是在整个基因组水平上，将产生巨大的洞察力， 个体基因以及通路的改变，并可能提供研究原因的新方法。 并揭示了新的治疗靶点。

Project Summary Alzheimer's disease (AD) is the most common cause of elderly dementia and currently there are no effective treatments. Selective neuronal vulnerability, appearance and accumulation of amyloid-β (Aβ) plaques, altered circuitry function, synaptic loss and degeneration are hallmarks of AD. Microglia and astrocytes, two major subtypes of glial cells, contribute to these neuropathological changes. However, the precise mechanisms controlling these processes remain elusive. One of the reasons may be that we still lack the information about the cell type-specific alterations and how they contribute to the onset and progression of AD. Here, we postulate that by revealing detailed changes in functional networks of genes in microglia and astrocytes, and how those processes intersect, we may be able to determine their contribution to neuropathological changes in AD. We will utilize Translational Ribosomal Affinity Purification combined with novel bioinformatics tools to construct functional network models of glial-specific functions and integrate these with AD quantitative genetic data. Using this framework, we will detect glial-specific genes most likely associated with AD pathology in an unbiased data-driven way and further investigate the functional network modules between astrocytes and microglia. Relevant candidate genes from this analysis will be examined in vitro for their role in AD progression, and emphasis will be given to cytokine-cytokine receptor pathways, especially interleukin 1 and colony stimulating factor 1 hubs. We will utilize primary cell cultures and co-cultures with astrocytes and microglia isolated from healthy and 5xFAD mice. Furthermore, we will investigate only genes that are relevant to the human pathology. To achieve this, we will test the appearance of AD hallmarks and/or the rescue phenotype in the microglia and astrocytes derived from the induced pluripotent stem cells (iPSC) isolated from AD patients and healthy controls. Finally, the most important gene candidates will be tested in vivo by gene expression manipulation in the appropriate mice lines. We will measure: i) Aβ accumulation, uptake, and degradation by microglia and astrocytes, ii) neuronal degeneration, iii) synaptic function, including synaptic markers and functional circuitry, iv) plaque formation, and v) cognitive function, assessed with a battery of behavioral tests. The proposed study will reveal the molecular profile of each cell type and examine their interaction. Generated datasets and bioinformatics tools will be shared with the public via web-based interface. These data will offer new insights into the appearance of the AD hallmarks and elucidate the basic mechanisms of the disease. Studying these changes, especially on the whole genome level, will engender tremendous insights into alterations in the individual genes, as well as pathways, and may offer new approaches to studying the cause of AD, as well as reveal novel therapy targets.