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.

项目概要/摘要 该项目将扩大 3D 电子显微镜的采集、重建、分析和传播 (3D EM) 参考数据，揭示了保存在大量遗产中的关键超微结构细节 这些样本最初是来自阿尔茨海默病 (AD) 患者的脑活检样本。 由神经病理学家 R.D. Terry 和 N. Gonatas（A. Einstein）收集、表征和存档 1960 年代），后来采集的样本作为脑脊液 (CSF) 药物输注研究的一部分 S. Mirra（位于 埃默里大学在 1980 年代），埃利斯曼、马斯利亚、特里和米拉在 1980 年代使用早期的方法重新审视了它们。 3D EM 方法，发现超微结构保存良好，显示成对的螺旋 细丝 (PHF) 和淀粉样蛋白积累以及亚细胞器和细胞骨架的修饰 在这里，我们将利用高通量方面的最新进展， 自动化 3D EM 可大规模检查这些珍贵样本，重建数百个大脑 有或没有 PHF 的细胞，通过更大的脑容量追踪轴突（并绘制神经胶质细胞和突触图） 我们的目标是针对与斑块和缠结相关的区域，重点关注。 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 一些人怀疑这是 AD 发生和发展的基础。 体细胞/树突状变性的进展，我们将针对可操作定义的代表进展的细胞 AD 中所见的神经衰退；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表征联系起来 同样，我们将分析斑块内和附近轴突变性的进展作为数据。 获得的结果表明轴突可能在其母体细胞体和树突之前变得营养不良 退化。