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: 10432028
• 负责人: JUDITH FRYDMAN
• 金额: $ 25.07万
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10432028
• 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 总结： 该核心将开发工具来定义衰老过程中的蛋白质稳态网络（PN）， 疾病相关的聚集倾向蛋白，并将提供技术支持项目1 - 4。的 资源也将用于测试和验证核心D的化合物。核心B将有两个站点紧密相连 地理接近。斯坦福大学的Frydman小组将建立分子传感器和测定来测量 老化过程中蛋白质稳态的变化，从为酵母伴侣机制开发的报告分子开始， 泛素-蛋白酶体系统（UPS）。Gladstone/UCSF的Finkbeiner小组将建立在哺乳动物的基础上， 表达和分析工具，以评估蛋白质聚集和自噬，并帮助PL解决特定的 他们的多路纵向单细胞成像平台至关重要。通过携手合作- 测试传感器在所有人使用的实验中报告不同蛋白酶抑制事件的能力 项目，我们将产生一套分析，提供一个全面的看法PN的任何细胞。工具将 提供给研究界。核心B的主要任务是开发分子传感器， 稳健和定量地报告PN的每个分支以及老化和 疾病相关的错误折叠蛋白质。成像传感器也将使我们能够研究蛋白质稳态是如何 机制是从一个细胞传递到另一个细胞的。这些问题的答案将提供关键的见解 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， 用于保持健康和治疗疾病。我们建议：（1）开发和验证一个小组， 蛋白酶抑制传感器和报告器，以及（2）协助项目负责人（PL）分析PN 传感器：对任何细胞的PN状态进行全面检查。 我们的最终目标是建立一套全面的传感器兼容的384孔板，可以使用 作为一个综合小组来询问PN的所有方面。通过综合知识的集体主体， 核心B将产生一个新的蛋白质稳定网络， 这是整个蛋白质稳定领域的宝贵工具箱，将促进进展，不仅在相关的实验室 除了PPG之外，其他实验室也对将蛋白质稳态功能障碍与衰老和疾病联系起来感兴趣。