Building a Toolbox of Sensors and Approaches to Monitor the Proteostasis Network Core B

构建监测蛋白质稳态网络核心 B 的传感器和方法工具箱

• 批准号: 10183111
• 负责人: JUDITH FRYDMAN
• 金额: $ 25.36万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10183111
• 关键词: Address; Affect; Aging; Autophagocytosis; Biological Assay; Biological Models; Biophysics; Biosensor; Cells; Collaborations; Communities; Data; Disease; Event; Fluorescence; Functional disorder; Generations; Geography; Goals; Health; Human; Image; Image Analysis; Investigation; Knowledge; Link; Literature; Measures; Modality; Molecular; Molecular Chaperones; Monitor; Mus; Neurodegenerative Disorders; Pathway interactions; Performance; Protein Biochemistry; Proteins; Proteomics; Quality Control; Reporter; Reporting; Research; Research Personnel; Resources; Site; System; Testing; Therapeutic; Time; Tissues; Ubiquitin; Work; Yeast Model System; Yeasts; base; biological adaptation to stress; cellular imaging; chaperone machinery; design; experience; experimental study; imaging platform; improved; insight; interest; misfolded protein; multicatalytic endopeptidase complex; novel; novel strategies; optical sensor; programs; protein aggregation; protein folding; proteostasis; quantitative imaging; sensor; single cell analysis; small molecule; success; synergism; tool; transcriptome sequencing; vector

Core B – Frydman/Finkbeiner - Sensors and Tools for Proteostasis Analyses Summary: This Core will develop tools to define the proteostasis network (PN) during aging and in the context of disease-associated aggregation-prone proteins and will provide technical support to Projects 1–4. The resources will also be used to test and validate compounds from Core D. Core B will have two sites in close geographical proximity. The Frydman group at Stanford will build molecular sensors and assays to measure proteostasis changes during aging, starting with reporters developed for yeast chaperone machinery and the ubiquitin-proteasome system (UPS). The Finkbeiner group at Gladstone/UCSF will build on mammalian expression and analysis tools to assess protein aggregation and autophagy and help PLs address specific questions for which their multiplexed longitudinal single-cell imaging platform is critical. By working hand-in- hand and testing the ability of the sensors to report on different proteostasis events in experiments used by all Projects, we will produce a set of assays that provide a comprehensive view of the PN of any cell. Tools will be available to the research community. A primary task of Core B will be to develop molecular sensors that robustly and quantitatively report on each branch of the PN and that change in aging and in the context of disease-associated misfolded proteins. Imaging sensors will also allow us to examine how proteostasis mechanisms are communicated from one cell to another. Answers to these questions will provide key insights into mechanisms of proteostasis dysfunction and protein aggregation during aging and will open new avenues for maintaining health and treating disease. We Propose to: (1) Develop and validate a panel of proteostasis sensors and reporters and (2) Assist project leaders (PLs) in the analysis of the PN sensors: towards a comprehensive exam of the PN status of any cell. Our ultimate goal is to build a comprehensive set of sensors compatible with 384-well plates that could be used as an integrated panel to interrogate all aspects of the PN. By synthesizing the collective body of knowledge of the proteostasis network into a set of fluorescence-based sensors and reporters, Core B will produce an invaluable toolbox for the whole proteostasis field that will facilitate progress, not only in the labs associated with this PPG but also in other labs interested in linking proteostasis dysfunction with aging and disease.

核心 B – Frydman/Finkbeiner - 用于蛋白质稳态分析的传感器和工具 概括： 该核心将开发工具来定义衰老过程中和在以下情况下的蛋白质稳态网络（PN）： 疾病相关的易聚集蛋白，将为项目 1-4 提供技术支持。这 资源还将用于测试和验证 Core D 的化合物。Core B 将有两个靠近的站点 地理位置接近。斯坦福大学的弗里德曼小组将构建分子传感器和检测方法来测量 衰老过程中蛋白质稳态发生变化，从为酵母伴侣机械开发的报告基因和 泛素蛋白酶体系统（UPS）。格拉德斯通/加州大学旧金山分校的 Finkbeiner 小组将以哺乳动物为基础 表达和分析工具可评估蛋白质聚集和自噬并帮助 PL 解决特定问题 他们的多重纵向单细胞成像平台对于解决这些问题至关重要。通过携手合作—— 手并测试传感器在所有人使用的实验中报告不同蛋白质稳态事件的能力 在项目中，我们将制作一套检测方法，提供任何细胞 PN 的全面视图。工具将是 可供研究界使用。 Core B 的首要任务是开发分子传感器 稳健、定量地报告 PN 的每个分支以及老龄化和老龄化背景下的变化 与疾病相关的错误折叠蛋白质。成像传感器还将使我们能够检查蛋白质稳态如何 机制从一个细胞传递到另一个细胞。这些问题的答案将提供关键见解 研究衰老过程中蛋白质稳态功能障碍和蛋白质聚集的机制，并将开辟新的途径 用于维持健康和治疗疾病。我们建议：（1）开发并验证一个小组 蛋白质稳态传感器和报告器，以及 (2) 协助项目负责人 (PL) 分析 PN 传感器：全面检查任何细胞的 PN 状态。 我们的最终目标是构建一套与 384 孔板兼容的全面传感器，可用于 作为一个综合小组来询问 PN 的各个方面。通过综合知识的集体 将蛋白质稳态网络转化为一组基于荧光的传感器和报告器，Core B 将产生一个 整个蛋白质稳态领域的宝贵工具箱将促进进步，而不仅仅是在相关实验室中 除了这个 PPG 之外，其他实验室也对蛋白质稳态功能障碍与衰老和疾病之间的联系感兴趣。