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。目标是 本项目的主要目的是阐明导致选择细胞脆弱性和致病的分子机制。 通过应用空间方法对阿尔茨海默病患者组织中高度多元化的RNA和蛋白质进行分析 横断面。我们将优化和规范多重荧光原位杂交平台。 检查来自单核转录组分析的AD相关基因集，以及 免疫组织化学(IHC)检测不同细胞类型人群中的病理蛋白标记物 完整的人脑组织。然后，这些方法将应用于脑部的阿尔茨海默病组织 了解疾病严重程度不同的地区1)的空间组织 这些脑区转录定义的细胞类型和疾病时该组织的变化 严重程度(例如选择性易损性或增殖)，2)阿尔茨海默病的分子通路被破坏 特定细胞类型的上下文，以及3)这些受影响的细胞类型与 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。这些数据生成 由mFISH和IHC共同提供了一张高分辨率的地图，展示了 临床上典型的阿尔茨海默氏症和对阿尔茨海默氏症病理的增强观察，以及强大的 未来对更大和更多样化的队列进行分析的技术平台。

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.