Temporal, Spatial and Cellular Dynamics of Amyloid Plaque Deposition

项目摘要 淀粉样斑块是阿尔茨海默病(AD)的典型病理特征之一。 淀粉样蛋白假说是淀粉样蛋白Aβ致病机制的标准模型。 过去25年的药物发现，导致了许多临床失败，包括 使接受治疗的患者的认知能力比接受安慰剂治疗的对照组更差。阿杜卡努单抗 (Aduhelm)是FDA最近批准的第一种基于 它能够促进淀粉样斑块的去除，表明这些 严苛。在AD中有几种不同类型的斑块和淀粉样沉积， 包括弥漫性、“经典”、“致密核”、神经炎斑块和脑血管淀粉样变性(CVA)。 和神经元内淀粉样蛋白沉积。虽然有很多关于淀粉样斑块的知识 AD的形态和成分，对斑块沉积的机制知之甚少，它们的 动力学和相互关系，不同类型细胞对其形成的贡献以及 它们在AD发病机制中的意义。我们试图研究淀粉样蛋白的这些关键方面。 通过标记特定细胞的蛋白质组，以详细和公正的方式进行斑块沉积 类型与非规范氨基酸，叠氮鸟亮氨酸，并在并入 ANL标记的蛋白质转化为淀粉样蛋白沉积。这项提案的目标是确定 用前沿双正交法研究淀粉样蛋白沉积的时间、空间和细胞动力学 非规范氨基酸标签(BONCAT)技术和特定的Cre驱动小鼠品系 细胞特异性合成可点击蛋白质，利用点击化学进行荧光 定位(FUNCAT)、纯化和浓缩以及生化分析。我们的中央 假说是不同类型的斑块在不同的时间由不同的机制沉积 不同种群神经元的时间和位置以及其中一些类型的斑块 可能在机制上与其他类型的淀粉样蛋白无关，并且可能是不同的 与发病机制和神经元变性有关。

Project Summary Amyloid plaques are one of the canonical pathological hallmarks of Alzheimer's disease (AD). The amyloid hypothesis is a standard model for amyloid Abeta pathogenesis that has driven drug discovery for the past 25 years, giving rise to many clinical failures, including drugs that make the treated patients cognitively worse than the placebo-treated controls. Aducanumab (Aduhelm) is the first disease modifying treatment recently approved by the FDA on the basis of its ability to facilitate the removal of amyloid plaques, indicating the importance of these strictures. There are several different types of plaques and amyloid deposits known in AD, including diffuse, “classical” “dense core”, neuritic plaques and cerebrovascular amyloid (CVA) and intraneuronal amyloid deposits. While there is much that is known about amyloid plaque morphology and composition in AD, less is known about mechanisms of plaque deposition, their dynamics and interrelationships, the contributions of different cell types to their formation and their significance for AD pathogenesis. We seek to investigate these critical aspects of amyloid plaque deposition in a detailed and un unbiased fashion by labeling the proteome of specific cell types with the non-canonical amino acid, azidonorleucine, and following the incorporation of ANL-labeled proteins into amyloid deposits. The goal of this proposal is to determine the temporal, spatial and cellular dynamics of amyloid deposition using cutting-edge biorthogonal non-canonical amino acid tagging (BONCAT) technology and specific Cre driver mouse lines for cell specific synthesis of clickable proteins, exploiting click chemistry for fluorescence localization (FUNCAT), purification and enrichment and biochemical analysis. Our central hypothesis is that different types of plaques are deposited by different mechanisms at different times and locations by different populations neurons and that some of these types of plaques may be mechanistically unrelated to the other types of amyloid and may be differentially associated with pathogenesis and neuronal degeneration.