UTSW-UNC Center for Cell Signaling Analysis

UTSW-UNC 细胞信号分析中心

• 批准号: 10705616
• 负责人: Gaudenz Danuser
• 金额: $ 107.41万
• 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/10705616
• 关键词: 3-Dimensional; Address; Adopted; Automobile Driving; Behavior; Biochemical; Biological; Biological Models; Biological Process; Biomedical Technology; Biosensor; Cell Separation; Cell Signaling Process; Cell physiology; Cells; Collaborations; Communities; Complex; Computer Models; Computer Vision Systems; Computer software; Core Facility; Data; Democracy; Discipline; Drug resistance; Ecosystem; Educational process of instructing; Environment; Etiology; Event; Feedback; Fiji; Genetic; Goals; Home; Image; Image Analysis; Imaging technology; Immune; Infrastructure; Institution; Invaded; Kinetics; Lateral; Libraries; Light; Malignant Neoplasms; Metabolism; Methods; Microscope; Microscopy; Modeling; Molecular; Molecular Analysis; Molecular Probes; Optics; Output; Pattern; Persons; Plug-in; Positioning Attribute; Postdoctoral Fellow; 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; 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; model organism; 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 细胞行为受到多种信号机制的调节，这些信号机制通常取决于分子动力学 在空间和时间上精确组织。分子事件的瞬态定位和动力学大量丢失 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 活细胞和组织中分子事件的定量分析，现在可以通过结合分子 探针设计、高分辨率活细胞显微镜和计算图像分析。然而，结合这些 将多种技术整合为一个工作整体，用于活细胞信号网络的定量分析仍然存在 具有挑战性的。拟议的细胞信号分析中心旨在使先进方法民主化 将这些工具交给那些没有将自己的职业生涯奉献给成像的科学家手中。我们建议开发 并传播一个用户友好的集成管道，该管道结合了 1) 生物传感器、光遗传学和 化学遗传学 2) 模块化、高速、高分辨率光片显微镜，以及 3) 图像分析和 计算建模以得出信号网络架构，包括因果关系和动力学 连接。该中心将集中精力整合 3 个技术开发项目： i) 基于具有互补性的替代方法的光遗传学、化学遗传学和生物传感器 能力并减少信号干扰。 ii) 多个模块化、经济高效、高分辨率的 3D 光片显微镜 由非专家快速组装，并提供约 220 和约 450 nm 的横向和轴向分辨率。这些 将被设计为最大限度地利用计算机视觉工作流程、生物传感器和 光遗传学/化学遗传学。 iii) 用于分析 2D 亚细胞信号转导的基于斐济的开源插件库 和 3D 活细胞延时数据，使用统计时间序列分析中的先进方法。 通过我们的协作驱动生物项目（DBP），我们将迭代地完善和改进我们的探针， 成像平台和图像分析方法。我们已经确定了 8 个解决免疫细胞主题的 DBP 迁移、免疫突触功能、机械感觉、肌节形成、癌症传播和 耐药性、突触可塑性和腹侧沟侵入。我们建议大力宣传 最大限度地利用现有基础设施的组件，包括拥有超过 3,000 个用户的成像设施 位于美国各地，广泛采用的 Fiji 软件生态系统、Addgene、GitHub 和 Applied 科学仪器。我们将提供广泛的培训（面对面、远程和主题驱动的课程）和 在一个全面且不断更新的网站上集中组织中心的产出。

项目成果

Filopodial protrusion driven by density-dependent Ena-TOCA-1 interactions.

由密度依赖性 Ena-TOCA-1 相互作用驱动的丝状突起。

• 发表时间: 2024-03-15
• 影响因子: 4
• 作者: Blake, Thomas C A;Fox, Helen M;Urbančič, Vasja;Ravishankar, Roshan;Wolowczyk, Adam;Allgeyer, Edward S;Mason, Julia;Danuser, Gaudenz;Gallop, Jennifer L
• 通讯作者: Gallop, Jennifer L