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

阿尔茨海默病选择性神经元脆弱性的代谢和蛋白质组学贡献者的质谱和多重免疫荧光成像

基本信息

批准号：
10704682
负责人：
PARAG Kumar MALLICK
金额：
$ 86.05万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2027-05-31
项目状态：
未结题

项目摘要

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.

我们的目标是揭示导致区域脆弱性的代谢和蛋白质信号传导途径阿尔茨海默氏病的新皮质锥体神经元，并确定新的检测目标和干涉。我们将比较前额皮质，这是一个受神经病理学影响的新皮质大脑区域在阿尔茨海默病的早期，初级视觉皮层是一个相对幸存的大脑区域。我们将使用质谱成像结合分割分析来识别空间变化从这些新皮质区域制备的死后脑切片中的小分子和蛋白质与对照组相比，处于阿尔茨海默病各个临床和病理阶段的捐赠者。然后，我们将在同一样本的连续切片上应用多重免疫荧光成像，以获取有关脆弱新皮质及其周围细胞和微环境变化的信息疾病进展期间的神经元，与临床严重程度、程度和位置相关神经病理变化和风险基因型。此外，我们将注册来自两个成像的数据识别区域、层状和层状代谢途径和蛋白质信号变化的技术细胞水平并定位导致阿尔茨海默病的分子和细胞表型的共变疾病脆弱性。除了生成细胞和分子的综合数据集阿尔茨海默病各个阶段的变化，这些研究将涉及开发、验证、和传播用于分析使用两种强大的成像生成的大型数据集的新工具工具，一种可以检测数百种分析物并有可能检测以前未知的分析物的工具一种是疾病的促成因素，另一种是通过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实验确定的代谢途径的细胞类型特异性靶向来修改治疗模型，例如用人类神经元或人源化小鼠复制皮质分层的大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。