Investigating FGF Signaling Dynamics in migrating cells

研究迁移细胞中的 FGF 信号动力学

• 批准号: 10679898
• 负责人: Theresa Gibney
• 金额: $ 4.47万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679898
• 关键词: Animals; Architecture; Behavior; Biological; Biosensor; Caenorhabditis elegans; Cells; Cellular biology; Chemotactic Factors; Communication; Cues; Cytoskeletal Modeling; Cytoskeletal Proteins; Cytoskeleton; Development; Disease; Extracellular Protein; FGF1 gene; Fibroblast Growth Factor; Fibroblast Growth Factor 8; Fibroblast Growth Factor Receptors; Fluorescence Recovery After Photobleaching; Genes; Genetic Transcription; Genome engineering; Goals; Growth Factor; Homologous Gene; Image; Ligands; MAP Kinase Gene; Malignant Neoplasms; Methods; Microscopy; Modeling; Molecular; Muscle; Muscle satellite cell; Myoblasts; Neoplasm Metastasis; Organism; PIK3CG gene; Pathway interactions; Postembryonic; Process; Proliferating; Proteins; Receptor Activation; Research; Resolution; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Site; Source; Spectrum Analysis; System; Tail; Testing; Time; Translating; Visualization; Work; cell behavior; cell motility; experimental study; extracellular; human disease; imaging approach; in vivo; in vivo Model; insight; migration; novel; optogenetics; permissiveness; progenitor; receptor; recruit; sex; tool

Project Summary Signaling dynamics are tightly regulated in space and time to control cell migration, proliferation, and differentiation. A comprehensive understanding of how extracellular inputs influence downstream signaling and cell behaviors will first require the ability to visualize key endogenous signaling proteins, cell architectures, and cytoskeletal regulators in vivo. Second, it will be essential to utilize precise tools to manipulate protein localization and signaling activity to reveal mechanisms. To explore how secreted signaling proteins disperse and modulate cell behavior, we are using a type of C. elegans muscle progenitors (SMs) as a tractable model for in vivo cell biology. The SMs migrate from near the tail of the worm to the center during larval development and are a classical system to investigate cell migration mechanisms. SM migration requires Fibroblast Growth Factor (FGF) signaling, which has been hypothesized to act as a long-range chemoattractant. However, the key molecular and cell biological mechanisms that translate FGF signaling into directed migration are not known. FGFs are often thought of as diffusible signaling proteins that can form gradients through free, extracellular dispersal, but this phenomenon has never been demonstrated for an endogenous FGF. To investigate how FGF proteins move between cells to regulate SM migration, I have tagged the endogenous FGF ligand and receptor involved in SM migration with fluorescent proteins and imaged their localization in vivo. Unexpectedly, I did not observe a gradient of FGF protein during SM migration, but instead observed low levels of FGF expression in a line of ventral midline cells that are located near migrating SMs. The proposed work builds on these findings and other new tools that I have made to visualize and manipulate key FGF pathway proteins in order to understand how FGF signaling regulates cell migration. Aim 1 will characterize endogenous FGF dynamics in a living animal and test how FGF moves between cells in vivo using genome engineering and live imaging approaches. Aim 2 will investigate how migrating cells respond to extracellular FGF by testing the extent to which FGF is a permissive or instructive signal, characterizing spatial organization of intracellular signaling downstream of FGF, and testing functions for localized pathway activation using optogenetics. These experiments will provide novel insights into how secreted signaling proteins move between cells and how migrating cells interpret and respond to dynamic extracellular cues.

项目摘要 信号传导动态在控制细胞迁移，增殖和 分化。对细胞外输入如何影响下游信号和 细胞行为将首先需要能够可视化关键的内源信号蛋白，细胞体系结构和 体内细胞骨架调节剂。其次，使用精确的工具来操纵蛋白质至关重要 定位和信号活性以揭示机制。探索分泌的信号蛋白如何分散 并调节细胞行为，我们使用一种类型的秀丽隐杆线虫肌肉祖细胞（SMS）作为可拖动模型 用于体内细胞生物学。在幼虫发育期间，SMS从蠕虫的尾巴附近迁移到中心 并且是研究细胞迁移机制的经典系统。 SM迁移需要成纤维细胞生长 因子（FGF）信号传导，该信号已被认为是远距离趋化剂。但是， 将FGF信号转化为定向迁移的关键分子和细胞生物学机制不是 已知。 FGF通常被认为是可扩散的信号蛋白，可以通过自由形成梯度， 细胞外散布，但是这种现象从未证明是内源性FGF的。到 研究FGF蛋白如何在细胞之间移动以调节SM迁移，我已经标记了内源性 FGF配体和与荧光蛋白迁移的SM迁移的受体，并成像其定位 体内。出乎意料的是，我在SM迁移过程中没有观察到FGF蛋白的梯度，而是观察到低 位于迁移SMS附近的腹部中线细胞系中FGF表达的水平。提议 工作以这些发现和其他我为可视化和操纵关键FGF的新工具为基础 途径蛋白是为了了解FGF信号如何调节细胞迁移。 AIM 1将表征 活动物中的内源性FGF动力学，并测试FGF如何使用基因组在体内移动细胞之间 工程和现场成像方法。 AIM 2将研究迁移细胞如何对细胞外反应 FGF通过测试FGF是允许或具有启发性的信号的程度，表征空间组织 FGF下游的细胞内信号传导以及使用局部途径激活的测试功能 光遗传学。这些实验将提供有关分泌信号蛋白如何移动的新见解 在细胞和迁移细胞之间如何解释和响应动态细胞外提示。