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: 10343738
• 负责人: Mark H Ellisman
• 金额: $ 74.89万
• 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/10343738
• 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 该项目将扩大三维电子显微镜的获取、重建、分析和传播 (3D EM)参考数据，揭示保存在非凡的遗产集合中的关键超微结构细节 取自阿尔茨海默病(AD)患者的脑组织活检样本。这些样本最初是 由神经病理学家R.D.特里和N.Gonatas收集、表征和存档(在A.爱因斯坦的 S)，后来作为涉及S.Mirra(AT)的脑脊液药物输注研究的一部分 20世纪80年代的埃默里(S)。20世纪80年代，埃利斯曼、马斯利亚、特里和米拉使用早期的 3DEM方法显示，超微结构保存良好，呈成对螺旋状 细丝(PHF)和淀粉样蛋白的积累以及对亚细胞器和细胞骨架的修饰 胞体、轴突和树突。在这里，我们将利用高吞吐量方面的最新进展， 自动3D EM大规模检查这些珍贵的样本，重建100个大脑 有无PHF的细胞，通过更大的大脑体积追踪轴突(并映射胶质细胞和突触) 比以前可行的。我们的目标是瞄准与斑块和缠结相关的区域，关注 现有发现表明细胞和网络易受攻击并包含分子相互作用的位置 一些人怀疑这是AD的发生和发展的基础。支持对 胞体/树突状退变的进展，我们将以手术定义的细胞为靶点来代表进展 如AD中所见的神经元衰退；测定胞浆中PHF的体积分数作为一种实用的分期 测量这一点，并将其与主要亚细胞微结构的数量变化的特征联系起来 选民。同样，我们将分析斑块内和斑块附近的轴突变性进程作为数据 研究表明，轴突可能在其母细胞体和树突之前变得营养不良。 堕落。