3D Reconstruction and Analysis of Alzheimer's Patient Biopsy Samples to Map and Quantify Hallmarks of Pathogenesis and Vulnerability

阿尔茨海默病患者活检样本的 3D 重建和分析，以绘制和量化发病机制和脆弱性的标志

• 批准号: 10115568
• 负责人: Mark H Ellisman
• 金额: $ 74.78万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10115568
• 关键词: 3-Dimensional; Acute; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Amyloid; Amyloid Fibrils; Archives; Area; Award; Axon; Back; Binding; Biochemical; Biopsy; Biopsy Specimen; Brain; Cataloging; Cell Nucleus; Cells; Cerebrospinal Fluid; Chromatin; Collection; Communities; Cytoplasm; Cytoskeletal Filaments; Cytoskeleton; Data; Data Analyses; Data Set; Dendrites; Electrons; Endoplasmic Reticulum; Funding; Future; Goals; Golgi Apparatus; Human; Infusion procedures; Investigation; Link; Literature; Location; Manuals; Maps; Measures; Methods; Microscopic; Microtubules; Mitochondria; Modification; Molecular Structure; Nerve Degeneration; Neurites; Neurofibrillary Tangles; Neuroglia; Neurons; Nuclear Pore; Organelles; Output; Parents; Pathogenesis; Patients; Performance; Pharmaceutical Preparations; Process; Production; Reporting; Request for Applications; Research; Research Personnel; Resolution; Running; Sampling; Scanning Electron Microscopy; Staging; Surveys; Synapses; Thick; Tissues; Training; United States National Institutes of Health; Work; axonal degeneration; base; brain cell; brain tissue; brain volume; cellular imaging; cellular targeting; convolutional neural network; data acquisition; data mining; deep learning; electron tomography; human model; image archival system; imaging Segmentation; insight; interest; microscopic imaging; mouse model; neuronal cell body; open source tool; paired helical filament; preservation; reconstruction; tomography; transmission process; web portal

PROJECT SUMMARY/ABSTRACT This project will expand the acquisition, reconstruction, analysis, and dissemination of 3D electron microscopic (3D EM) reference data, disclosing key ultrastructural details preserved within a remarkable collection of legacy biopsy brain samples from patients suffering from Alzheimer’s Disease (AD). These samples were originally collected, characterized and archived by neuropathologists R.D. Terry and N. Gonatas (at A. Einstein in the 1960’s), with later samples taken as part of a cerebrospinal fluid (CSF) drug infusion study involving S. Mirra (at Emory in the 1980’s). They were re-examined by Ellisman, Masliah, Terry, and Mirra in the 1980s, using early 3D EM methods, and were found to manifest excellent preservation of ultrastructure, showing paired helical filaments (PHF) and amyloid accumulations as well as modifications to subcellular organelles and cytoskeletons of the cell bodies, axonal and dendritic processes. Here, we will exploit recent advances in high throughput, automated 3D EM to massively scale the examination of these precious samples, reconstructing 100s of brain cells with and without PHF, tracking axons (and mapping glia and synapses) through much greater brain volumes than was feasible previously. Our goal is to target areas associated with both plaques and tangles, attending to locations where existing findings suggest cell and network vulnerability and contain molecular interactions suspected by some to underlie the initiation and progression of AD. Supporting investigations into the progression of soma/dendritic degeneration, we will target cells operationally defined to represent a progression of neuronal decline as seen in AD; determining the volume fraction of PHF in the cytoplasm as a practical staging measure and linking this to the characterization of quantitative changes in the microstructure of major subcellular constituents. Likewise, we will analyze the progression of axonal degeneration in and near plaques as data obtained suggests that axons may become dystrophic before their parent cell bodies and their dendrites degenerate.

项目摘要/摘要 该项目将扩大3D电子显微镜的获取，重建，分析和传播 （3D em）参考数据，披露保留在遗产的巨大收藏中的关键超微结构细节 来自阿尔茨海默氏病（AD）的患者的活检大脑样本。这些样品最初是 由神经病理学家R.D. Terry和N. Gonatas收集，特征和存档（在A. Einstein 1960年代），后来的样品是涉及链球菌的脑脊液（CSF）药物输注研究的一部分 埃默里（Emory）在1980年代）。他们在1980年代被Ellisman，Masliah，Terry和Mirra重新检查， 3D EM方法，并被发现表现出极好的超微结构保存，显示了配对的螺旋形 细丝（PHF）和淀粉样蛋白的积累以及对亚细胞细胞器和细胞骨架的修饰 细胞体，轴突和树突过程。在这里，我们将探讨高通量的最新进展， 自动化的3D em以大规模扩展这些珍贵样品的检查，重建100 s脑 具有和不具有PHF的细胞，通过更大的脑量跟踪轴突（以及映射神经胶质和突触） 比以前可行的。我们的目标是针对与斑块和缠结相关的区域 现有发现表明细胞和网络脆弱性并包含分子相互作用的位置 有人怀疑是AD的主动性和进步的基础。支持调查 SOMA/树突变性的进展，我们将以操作定义的细胞靶向代表进展 在AD中看到的神经元下降；确定细胞质中PHF的体积分数作为实际分期 测量并将其与主要亚细胞微观结构的定量变化的表征相关联 成分。同样，我们将分析斑块中轴突变性的进展 获得的表明，轴突在其母细胞体和树突之前可能会变成营养不良 退化。