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

基本信息

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
项目类别：
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

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

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项目摘要

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-传感器和用于蛋白质的工具分析概括：该核心将开发工具来定义衰老期间的蛋白质量表网络（PN）易于疾病相关的可聚集蛋白，并将为项目1-4提供技术支持。这资源还将用于测试和验证CoreD。CoreB的化合物地理邻近。斯坦福大学的Frydman组将建立分子传感器和测定法以测量衰老过程中的蛋白质静态变化，从为酵母伴侣机械开发的记者开始泛素 - 蛋白酶体系统（UPS）。 Gladstone/UCSF的Finkbeiner Group将以哺乳动物为基础评估蛋白质聚集和自噬的表达和分析工具，并帮助PL解决特定其多路复用纵向单细胞成像平台的问题至关重要。通过手工工作在所有人使用的实验中，手动和测试传感器报告不同蛋白抑制事件的能力项目，我们将制作一套测定法，从而为任何单元的PN提供全面的看法。工具将是可用于研究社区。核B的主要任务是开发分子传感器坚固，定量地报告PN的每个分支，以及在衰老的变化和与疾病相关的错误折叠蛋白。成像传感器还将使我们能够检查蛋白质的含量机制从一个单元格传达到另一个单元格。这些问题的答案将提供关键的见解进入衰老期间蛋白质的功能障碍和蛋白质聚集的机制，并将开放新的途径维持健康和治疗疾病。我们建议：（1）开发和验证一个面板 Proteostasis传感器和记者，（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。我们的最终目标是构建与384孔板兼容的全面传感器作为综合面板，询问PN的所有方面。通过综合集体知识的团体蛋白质的网络进入一组基于荧光的传感器和记者，Core B将产生一个整个蛋白质病领域的宝贵工具箱将促进进步，不仅在实验室相关的实验室有了这个PPG，还包括有兴趣将蛋白质功能障碍与衰老和疾病联系起来的其他实验室。