Cellular Anatomy of Tau Seeding

Tau 播种的细胞解剖学

• 批准号: 10575918
• 负责人: Sarah H Shahmoradian
• 金额: $ 24.6万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10575918
• 关键词: 3-Dimensional; Alzheimer' s Disease; Antibodies; Axon; Biosensor; Brain; Bypass; Cells; Cytoplasm; Data; Dendrites; Diamond; Electron Microscopy; Electrons; Event; Faculty; Human; Image; Immunofluorescence Immunologic; Impaired cognition; In Situ; Label; Light; Methods; Modeling; Molecular; Molecular Conformation; Molecular Machines; Monitor; Morphologic artifacts; Neurodegenerative Disorders; Neurons; Organelles; Pathologic; Pathology; Pattern; Presynaptic Terminals; Process; Proteins; Recombinants; Resolution; Resources; Role; Subcellular Anatomy; System; Techniques; Testing; Transgenic Mice; Visualization; human stem cells; imaging approach; induced pluripotent stem cell; light microscopy; live cell imaging; member; microscopic imaging; nanoscale; neuronal cell body; novel; protein aggregation; spatiotemporal; tau Proteins; tau conformation; tool; uptake

Project Summary/Abstract: The propagation of tau pathology along synaptically connected brain circuits is considered a major driver of cognitive decline in Alzheimer’s Disease (AD). However, the mechanisms underlying proteopathic tau seed entry and amplification within human neurons remain unclear. Given the putative role of brain circuits in tau propagation, axons/axon terminals or dendrites may be involved in tau seed uptake. However, this has never been determined in neuronal models, and typical methods to study these events, such as light microscopy and immunofluorescence alone, have critical resolution limits. Thus, we cannot yet hypothesize where in a neuron that the tau seeds access the cytoplasm and replicate. To answer these questions, we will apply high-resolution correlative light and electron microscopic imaging (3D-CLEM), combined with advanced cell-based fluorescent biosensors and novel conformational antibodies with live cell imaging in human neurons derived from inducible pluripotent stem cells (iPSCs) available here in our center. Our strategy should enable determining where tau seed uptake occurs, and the localization and sub-cellular components and/or molecular machinery involved in tau seed amplification, while bypassing protein aggregate extraction techniques that introduce experimental artifact. By exploring these mechanisms using high-resolution correlative imaging within human neurons, we will address how proteopathic seeds access the cytoplasm and replicate, which may have broader implications for other related neurodegenerative diseases.

项目摘要/摘要： tau病理沿突触连接的脑电路的传播被认为是 阿尔茨海默氏病认知能力下降的主要驱动力（AD）。但是，基础机制 人神经元内的蛋白质tau种子进入和扩增仍不清楚。给定推定 脑电路在tau繁殖，轴突/轴突末端或树突中的作用可能参与tau种子 吸收。但是，这从未在神经元模型和研究的典型方法中确定。 这些事件，例如光显微镜和免疫荧光，具有关键的分辨率限制。 那就是我们还不能假设在神经元中tau种子进入细胞质和 复制。要回答这些问题，我们将应用高分辨率纠正灯和电子 微观成像（3D-CLEM），结合了高级基于细胞的荧光生物传感器和新型 具有诱导多能的人类神经元中的活细胞成像的构象抗体 干细胞（IPSC）在我们的中心提供。我们的策略应确定tau种子在哪里 发生吸收，涉及的定位和亚细胞组件和/或分子机械 在tau种子扩增中，同时绕过引入的蛋白质骨料提取技术 实验工件。通过使用高分辨率正确的成像探索这些机制 人神经元，我们将解决蛋白质疗法如何进入细胞质并复制，这 可能对其他相关神经退行性疾病具有更广泛的影响。