Project 2: 3-D Molecular atlas of AD proteinopathy

项目2：项目摘要/摘要 阿尔茨海默病(AD)是一种最常见的神经退行性疾病，其结果有多种 症状，其中最突出的是进行性认知能力下降。而主要的病理改变是 与AD相关的蛋白病变是淀粉样β蛋白(斑块)和tau(神经原纤维)的聚集体 这些蛋白病变的外观、密度和分布显示出明显的异质性。 跨越个体和大脑区域。此外，个体经常表现出这两者同时出现 蛋白质病与其他蛋白质聚集体，如TDP43和α-突触核蛋白；后两种是 主要与肌萎缩侧索硬化症、额颞叶痴呆和 帕金森氏症，但其在AD中的发生并不罕见。从我们的块状和单核RNA-seq 对身体组织的研究，我们已经确定了特定的细胞特征/推测的亚型 丰度与缠结和斑块有关。然而，这些块状和单核实验 方法不会保留有关这些像元类型的定位(或不定位)方式的空间信息 病理性包裹体，从而产生一幅不完整的画面，说明老年人的细胞结构 在AD病理存在的情况下，大脑会发生改变。在这里，我们的目标是检查空间分布和 使用高度多元化的空间方法直接在完整的人脑组织中组织这些细胞类型。 我们提出了一种新的但明显可扩展的蛋白质和基因表达方法的组合，以 研究大量(300微米厚)的人体组织与这些蛋白质病的组合。 为了研究蛋白质聚集体对“高度局部性”的全面影响，这些大量的文献是必要的。 环境。具体地说，我们使用了高度多元化的抗体染色和全基因组 用空间转录学方法检验我们关于神经元和胶质细胞重组的假说 病理性包涵体周围的类型，重点是两种主要的AD相关蛋白病变(淀粉样蛋白 和tau)。最终，这将生成一份关于细胞类型和基因表达签名的高分辨率图谱 在阿尔茨海默病中常见的蛋白质聚集体的存在下发生变化的老化的人脑组织， 补充了许多现有的和正在进行的大量和单核RNA-SEQ研究 人类神经退行性变研究。

PROJECT 2: PROJECT SUMMARY/ABSTRACT Alzheimer’s Disease (AD) is the most prevalent neurodegenerative disease of aging, and results in a variety of symptoms, the most prominent of which is progressive cognitive decline. Whereas the primary pathological proteinopathies associated with AD are aggregates of amyloid-beta (plaques) and tau (neurofibrillary tangles), the appearance, density, and distribution of these proteinopathies shows substantial heterogeneity across individuals and brain regions. In addition, individuals often display co-occurrence of these two proteinopathies with other protein aggregates such as TDP43 and alpha-synuclein; these latter two are primarily associated with diseases such as Amyotrophic Lateral Sclerosis, Frontotemporal Dementia and Parkinson’s Disease, but their occurrence in AD is not rare. From our bulk and single-nucleus RNA-seq studies on post-mortem tissue, we have identified specific cellular signatures/putative subtypes whose abundance is associated with tangles and plaques. However, these bulk and single-nucleus experimental approaches do not preserve spatial information about how these cell types are localized (or not) around pathological inclusions, and thus generate an incomplete picture of how the cellular structure of the aged brain is altered in the presence of AD pathology. Here, we aim to examine the spatial distribution and organization of these cell types directly in intact human brain tissue using highly-multiplexed spatial methods. We propose a combination of novel but demonstrably scalable protein and gene expression approaches to study large volumes (300-micron thickness) of human tissue with combinations of these proteinopathies. These large volumes are necessary to study the full impact of protein aggregates on the “highly local” environment. Specifically, we use a combination of highly multiplexed antibody staining and genome-wide spatial transcriptomics methods to examine our hypothesis on the reorganization of neuronal and glial cell types around pathological inclusions, with a focus on the two major AD-associated proteinopathies (amyloid and tau). Ultimately, this will generate a high-resolution atlas of cell types and gene expression signatures in aged human brain tissue that are altered in the presence of protein aggregates commonly found in AD, complementing many of the existing and ongoing bulk and single-nucleus RNA-seq studies in the field of human neurodegeneration research.