Mass spectrometry and multiplexed immunofluorescence imaging of metabolic and proteomic contributors to selective neuronal vulnerability in Alzheimer's disease

我们的目标是揭示代谢和蛋白质信号通路有助于区域脆弱性， 阿尔茨海默病中的新皮层锥体神经元，并鉴定用于检测的新靶点， 干预我们将比较前额叶皮层，一个受神经病理学影响的新皮层大脑区域， 在阿尔茨海默病的早期，初级视觉皮层，一个相对备用的大脑区域。我们将 使用质谱成像结合分割分析，以确定空间变化 小分子和蛋白质在死后大脑切片中的含量， 与对照组相比，在阿尔茨海默病的各种临床和病理阶段的供体。然后我们 将对来自相同标本的连续切片进行多重免疫荧光成像， 获得关于脆弱的新皮层内和周围的细胞和微环境变化的信息 神经元在疾病进展过程中，与临床严重程度，程度和位置相关， 神经病理学改变和风险基因型。此外，我们将记录来自两个成像的数据 技术，以确定代谢途径和蛋白质信号转导的变化，在区域，层流和 细胞水平，并定位有助于阿尔茨海默氏症的分子和细胞表型的协变 疾病脆弱性。除了生成细胞和分子的全面数据集之外， 在阿尔茨海默病的各个阶段的变化，这些研究将涉及开发，验证， 和传播新的工具，用于分析使用两种强大的成像技术生成的大型数据集 工具，一个可以检测数百种分析物的工具， 另一种是利用一组选定的已知标记物提供更高的分辨率。 从长远来看，这些研究产生的数据可以为测试新疾病提供基础- 通过使用实验的细胞类型特异性靶向鉴定的代谢途径来修改治疗 模型，例如用人类神经元或人源化小鼠复制皮质层压的脑类器官 嵌合体来模拟神经元和非神经元细胞类型之间的相互作用。

We aim to uncover metabolic and protein signaling pathways contributing to the regional vulnerability of neocortical pyramidal neurons in Alzheimer's disease and to identify novel targets for detection and intervention. We will compare the prefrontal cortex, a neocortical brain area afflicted by neuropathology early in Alzheimer's disease, with the primary visual cortex, a brain area that is relatively spared. We will use mass spectrometric imaging in combination with segmentation analyses, to identify spatial changes of small molecules and proteins in postmortem brain sections prepared from these neocortical areas from donors at various clinical and pathological stages of Alzheimer's disease compared to controls. Then, we will apply multiplexed immunofluorescence imaging on sequential sections from the same specimens, to obtain information on cellular and microenvironment changes in and around vulnerable neocortical neurons during disease progression, correlated with the clinical severity, degree and location of neuropathological changes, and the risk genotype. Further, we will register the data from both imaging techniques to identify metabolic pathways and protein signaling changes at the regional, laminar and cellular level and to locate covariation in molecular and cellular phenotypes contributing to Alzheimer's disease vulnerability. In addition to generating a comprehensive dataset of the cellular and molecular changes at various stages of Alzheimer's disease, these studies will involve the development, validation, and dissemination of novel tools for analysis of large datasets generated using two powerful imaging tools, one that detects hundreds of analytes with the possibility of detecting previously unknown contributors to disease and the other that provides higher resolution with a select set of known markers. In the long term, the data generated from these studies could provide the basis for testing novel disease- modifying treatments by cell-type specific targeting of identified metabolic pathways using experimental models, such as brain organoids to replicate cortical lamination with human neurons or humanized mouse chimeras to model interactions between neurons and non-neuronal cell types.