Proteostasis in Aging and Neurodegenerative Disease

项目摘要 社会面临的最具挑战性的问题之一是寿命延长带来的混合好处，也就是 伴随着神经退行性疾病风险的增加。这项提议的一个中心主题是 衰老与蛋白质质量控制机制的能力下降有关，从而导致蛋白质 错误折叠和聚集，导致细胞和组织衰竭和神经退行性疾病。在这 PPG，我们已经组建了S.Finkbeiner(加州大学旧金山分校)、D.Finley(哈佛大学)、J.Frydman(斯坦福大学)、J. Kelly(斯克里普斯)和R.Morimoto(西北大学)研究衰老中的蛋白平衡失败作为基础 阿尔茨海默病、肌萎缩侧索硬化症、亨廷顿病患者Tau、SOD1和Expanded PolyQ的错误折叠和聚集 疾病和共济失调。这个PPG团队的一个独特优势是我们在多个方面的专业知识 生物系统包括酿酒酵母、线虫、小鼠、患者来源的细胞和分化的神经元， 以及来自生化和生物物理的多种实验方法，以及聚集的活细胞成像 表型和小分子高通量筛选。我们假设，方法的独特丰富性 将提供新的见解，揭示衰老是如何影响蛋白质平衡网络的 蛋白质合成和分子伴侣的(Pn)，运输机械和降解的手臂 PN由泛素-蛋白酶体和自噬溶酶体途径组成。了解如何 衰老在细胞、组织和组织水平上对PN的影响将提供对 蛋白失稳过程中促进和加速Tau、SOD1和Tau的聚集 膨胀的多聚Q蛋白导致AD和其他神经退行性疾病。通过四个项目和 四个核心，我们的团队将探索衰老如何影响分子伴侣的功能，从而影响新生- Tau和聚谷氨酰胺在酵母中的链合成和非路径聚集特性(Proj.1)和In 短寿和长寿线虫的不同组织。4)。我们将检查PN的退化臂 蛋白酶体活性水平改变的转基因小鼠及Tau和突变体对蛋白毒性的影响 SOD(项目2)，并通过监测自噬溶酶体在人IPSCs衍生的患者神经元中 路径(项目3)衰老和Tau、TDP43的表达。这些项目将得到以下方面的支持： 发展PN报告量化的协调管理核心A、蛋白质抑制传感器核心B 不同的PN活性、蛋白质平衡蛋白质组学核心C和蛋白质平衡调节剂药理核心D 开发一种小分子策略来恢复衰老和神经退行性疾病的PN功能。 通过合作，这些核心和项目将产生PN试剂和工具、数据集和小分子 量化和干扰PN的不同成分，以生成老化和 所有生物系统的神经退化，并开发一种小分子策略来防止或 逆转导致神经退行性疾病的PN中与年龄和疾病相关的失败。

Project Summary One of the most challenging problems facing society is the mixed benefit of longer lifespan that is also accompanied by the increased risk for neurodegenerative diseases. A central theme of this proposal is that aging is associated with the declining capability of the protein quality control machinery, leading to protein misfolding and aggregation, and resulting in cell and tissue failure and neurodegenerative disease. In this PPG, we have assembled the team of S. Finkbeiner (UCSF), D. Finley (Harvard), J. Frydman (Stanford), J. Kelly (Scripps) and R. Morimoto (Northwestern) to examine proteostasis failure in aging as the basis for misfolding and aggregation of Tau, SOD1 and expanded polyQ in Alzheimer's disease (AD), ALS, Huntington's disease and Ataxias, respectively. A distinctive strength of this PPG team is our expertise with multiple biological systems including S. cerevisiae, C. elegans, mice, patient derived cells and differentiated neurons, and multiple experimental approaches from biochemical and biophysical, live cellular imaging of aggregation phenotypes, and small molecule high-throughput screens. We posit that the unique richness of approaches afforded by this team will provide novel insights that will uncover how aging affects the proteostasis network (PN) of protein synthesis and molecular chaperones, transport machineries, and the degradative arms of the PN comprised of the ubiquitin-proteasome and autophagy lysosomal pathways. An understanding of how aging affects the PN at the cellular, tissue, and organismal level will provide a mechanistic understanding on the events during proteostasis failure that contributes to and accelerates aggregation of Tau, SOD1 and expanded polyQ proteins leading to AD and other neurodegenerative diseases. Through four Projects and four Cores, our team will explore how aging affects the function of molecular chaperones to influence nascent- chain synthesis and the off-pathway aggregation properties of Tau and polyglutamine in yeast (Proj. 1) and in different tissues of short-and-long-lived C. elegans (Proj. 4). We will examine the degradative arms of the PN in transgenic mice with altered levels of proteasome activity and the effects on proteotoxicity of Tau and mutant SOD1 (Proj. 2), and in human iPSCs derived patient neurons by monitoring the autophagic lysosomal pathways (Proj. 3) challenged by aging and expression of Tau or TDP43. These Projects will be supported by: the coordinating Administrative Core A, Proteostasis Sensors Core B that develops PN reporters to quantify different PN activities, Proteostasis Proteomics Core C, and the Proteostasis Regulator Pharmacology Core D to develop a small molecule strategy to restore PN functionality in aging and neurodegenerative disease. Working together, the Cores and Projects will generate PN reagents and tools, datasets and small molecules to quantify and perturb different components of the PN, to generate a comparative analysis of aging and neurodegeneration across all biological systems, and to develop a small molecule strategy to prevent or reverse the age-and-disease dependent failures in the PN leading to neurodegenerative disease.