Spatial analysis of regional, cell type and molecular hallmarks of Alzheimer's disease

摘要（项目3） 阿尔茨海默病细胞脆弱性的分子机制尚不清楚。客观 该项目的目的是阐明选择细胞脆弱性和发病机制的分子机制 通过应用空间方法对阿尔茨海默病影响组织进行高度多重RNA和蛋白质分析， 路段我们将优化和标准化多重荧光原位杂交（mFISH）平台 检查来自单核转录组分析的AD连锁基因集，以及 免疫组化（IHC）检查病理蛋白标志物的细胞类型群体的背景下， 完整的人脑组织然后将这些方法应用于来自大脑的阿尔茨海默病组织， 受不同疾病严重程度影响的地区，以了解1） 转录组学定义的细胞类型在这些大脑区域和变化，在这个组织与疾病 严重性（例如，选择性脆弱性或增殖），2）阿尔茨海默病中的分子途径被破坏， 特定细胞类型的背景，和3）这些受影响的细胞类型和细胞的标记物之间的关系。 阿尔茨海默病的病理，如淀粉样前体蛋白和过度磷酸化的tau蛋白。这些数据生成 通过mFISH和IHC一起将提供高分辨率的细胞和分子结果图， 临床上典型的阿尔茨海默病和阿尔茨海默病病理学的增强视图，以及强大的 为今后分析更大和更多样化的群组提供技术平台。

ABSTRACT (PROJECT 3) The molecular mechanisms underlying cellular vulnerability in Alzheimer’s disease are unclear. The objective of this project is to clarify the molecular mechanisms responsible for select cell vulnerability and pathogenesis by applying spatial methods for highly multiplexed RNA and protein analysis in Alzheimer’s-affected tissue sections. We will optimize and standardize platforms for multiplexed fluorescence in situ hybridization (mFISH) to examine AD-linked gene sets derived from single nucleus transcriptome analysis, as well as immunohistochemistry (IHC) to examine pathological protein markers in the context of cell type populations in intact human brain tissue. These methods will then be applied to Alzheimer’s disease tissues from brain regions affected in different levels of disease severity to understand 1) the spatial organization of transcriptomically-defined cell types in these brain regions and changes in this organization with disease severity (e.g. selective vulnerability or proliferation), 2) molecular pathways disrupted in Alzheimer’s disease in the context of specific cell types, and 3) the relationship between these affected cell types and the markers of Alzheimer’s pathology, such as amyloid precursor protein and hyperphosphorylated tau. These data generated by mFISH and IHC together will provide a high-resolution map of the cellular and molecular consequences of clinically typical Alzheimer’s disease and an enhanced view of Alzheimer’s pathology, as well as a powerful technology platform for future analyses of larger and more varied cohorts.