UTSW-UNC Center for Cell Signaling Analysis

UTSW-UNC 细胞信号分析中心

• 批准号: 10412148
• 负责人: Gaudenz Danuser
• 金额: $ 160.71万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-17 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10412148
• 关键词: 3-Dimensional; Address; Adopted; Animal Model; Automobile Driving; Behavior; Biochemical; Biological; Biological Models; Biological Process; Biomedical Technology; Biosensor; Cell Signaling Process; Cell physiology; Cells; Collaborations; Communities; Complex; Computer Models; Computer Vision Systems; Computer software; Core Facility; Data; Discipline; Drug resistance; Ecosystem; Environment; Etiology; Event; Feedback; Fiji; Genetic; Goals; Home; Image; Image Analysis; Imaging technology; Immune; Infrastructure; Institution; Kinetics; Lateral; Libraries; Light; Malignant Neoplasms; Metabolism; Methods; Microscope; Microscopy; Modeling; Molecular; Molecular Analysis; Molecular Probes; Optics; Output; Pattern; Persons; Positioning Attribute; Process; Proteins; Proteomics; Reporter; Research Project Grants; Resolution; Sarcomeres; Scheme; Scientist; Series; Signal Transduction; Signal Transduction Pathway; Speed; Standardization; Statistical Methods; Synaptic plasticity; Technology; Testing; Time; Time Series Analysis; Tissues; Trainers Training; Training; United States; Update; Variant; Visualization; analog; base; biological systems; career; cell behavior; cell motility; computational pipelines; cost effective; design; field study; imaging facilities; imaging modality; imaging platform; imaging probe; immunological synapse; immunological synapse formation; improved; instrumentation; live cell microscopy; mechanotransduction; migration; molecular dynamics; molecular imaging; molecular modeling; multiplex detection; network architecture; neutrophil; open source library; optogenetics; predictive modeling; programs; spatiotemporal; synaptic function; technology development; temporal measurement; tool; user-friendly; web site

ABSTRACT Cellular behavior is regulated by diverse signaling mechanisms that often depend on molecular dynamics precisely organized in space and time. The transient positioning and kinetics of molecular events is lost in bulk biochemical analysis and in single cell proteomics. Understanding such factors requires visualization and quantitative analysis of molecular events in living cells and tissues, now made possible by combining molecular probe design, high-resolution live cell microscopy and computational image analysis. However, combining these diverse technology into a working whole for the quantitative analysis of live cell signaling networks remains challenging. The proposed Center for Cell Signaling Analysis aims to democratize advanced methods that put these tools in the hands of scientists who have not devoted their careers to imaging. We propose to develop and disseminate a user-friendly and integrated pipeline that combines 1) biosensors, optogenetics and chemogenetics 2) modular, high-speed, and high-resolution light-sheet microscopes, and 3) image analysis and computational modeling to derive signaling network architecture, including the causality and kinetics of connections. The Center will concentrate its integration effort between 3 Technology Development Projects: i) Optogenetics, chemogenetics, and biosensors based on alternate approaches with complementary capabilities and reduced perturbation of signaling. ii) Multiple modular, cost-effective, and high-resolution 3D light-sheet microscopes that can be assembled rapidly by non-experts and deliver ~220 and ~450 nm lateral and axial resolution. These will be designed to maximally leverage computer vision workflows, biosensors, and optogenetics/chemogenetics. iii) A library of open-source Fiji-based plugins for the analysis of subcellular signal transduction in 2D and 3D live cell time-lapse data using advanced methods in statistical time series analysis. Through our collaborative Driving Biological Projects (DBPs), we will iteratively refine and improve our probes, imaging platforms and image analysis methods. We have identified 8 DBPs that address topics in immune cell migration, immunological synapse function, mechanosensation, sarcomere formation, cancer dissemination and drug resistance, synaptic plasticity, and ventral furrow ingression. We propose a strong dissemination component that maximally leverages existing infrastructure, including imaging facilities with >3,000 users located throughout the United States, the widely adopted Fiji software ecosystem, Addgene, GitHub, and Applied Scientific Instrumentation. We will provide extensive training (in person, remote, and topic-driven courses) and centrally organize the Center outputs on a comprehensive and continuously updated website.

抽象的 细胞行为受到各种信号传导机制的调节，这些信号传导机制通常取决于分子动力学 精确组织在时空。分子事件的瞬态定位和动力学在大量中丢失 生化分析和单细胞蛋白质组学。了解这些因素需要可视化和 对活细胞和组织中分子事件的定量分析，现在通过结合分子而成为可能 探针设计，高分辨率活细胞显微镜和计算图像分析。但是，将这些结合在一起 多样化的技术成为一个工作整体，用于定量分析活细胞信号网络 具有挑战性的。拟议的细胞信号分析中心旨在使高级方法民主化 将这些工具掌握在没有专门职业成像的科学家手中。我们建议发展 并传播结合用户友好和集成的管道1）生物传感器，光遗传学和 化学遗传学2）模块化，高速和高分辨率灯页显微镜，以及3）图像分析和 计算建模以得出信号网络体系结构，包括因果关系和动力学 连接。该中心将集中于3个技术开发项目之间的集成工作： i）基于互补的替代方法的光遗传学，化学遗传学和生物传感器 功能和信号传导的扰动降低。 ii）多个模块化，成本效益和高分辨率3D灯页显微镜可以是 通过非专家迅速组装，并提供〜220和〜450 nm的横向和轴向分辨率。这些 旨在最大程度地利用计算机视觉工作流，生物传感器和 光遗传学/化学遗传学。 iii）一个开源斐济插件库，用于分析2D中的亚细胞信号转导 在统计时间序列分析中使用高级方法和3D实时延时数据。 通过我们的协作驾驶生物项目（DBP），我们将迭代完善并改善我们的探针， 成像平台和图像分析方法。我们已经确定了8个DBP，可以解决免疫细胞中的主题 迁移，免疫突触功能，机械敏化，肌节形成，癌症传播和 耐药性，突触可塑性和腹侧犁沟入口。我们提出了强烈的传播 最大程度地利用现有基础架构的组件，包括> 3,000个用户的成像设施 广泛采用的斐济软件生态系统，Addgene，github和Appliped，位于整个美国。 科学仪器。我们将提供广泛的培训（亲自，远程和主题驱动的课程）和 在全面且不断更新的网站上中心组织中心输出。