Coordinated Actin Regulation in Directed Neural Crest Cell Migration

定向神经嵴细胞迁移中的协调肌动蛋白调节

• 批准号: 10613445
• 负责人: Shuyi Nie
• 金额: $ 31.08万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10613445
• 关键词: Actins; Affect; Biological; Cell Culture Techniques; Cell Migration Inhibition function; Cell Polarity; Cell membrane; Cell model; Cell physiology; Cells; Collaborations; Complex; Computer Models; Computer Simulation; Congenital Abnormality; Craniofacial Abnormalities; Cytoplasm; Cytoskeletal Modeling; Cytoskeleton; Data; Defect; Development; Disease; Embryonic Development; Equilibrium; Etiology; Event; Experimental Designs; Family; Feedback; Filament; Filopodia; Goals; Guanosine Triphosphate Phosphohydrolases; Heart Abnormalities; Homeostasis; Human; Imaging technology; Knowledge; Lead; Malignant Neoplasms; Mediating; Membrane; Microfilaments; Modeling; Molecular; Morphogenesis; Neural Crest; Neural Crest Cell; Polymers; Process; Proteins; Public Health; Regulation; Research; Role; Signal Transduction; Stress Fibers; System; Testing; Tissues; Work; Xenopus; cdc42 GTP-Binding Protein; cell behavior; cell motility; experimental study; human disease; improved; in silico; in vivo; migration; models and simulation; neurodevelopment; novel; polarized cell; polymerization; recruit; rho; scaffold

PROJECT SUMMARY Cell migration is critical to development and disease, and yet yet our understanding for this complex dynamic event is rather limited. Cell culture studies have uncovered crucial regulators of actin cytoskeleton, including the Rho family of GTPases (RhoA, Rac1, and Cdc42), that influence particular processes of cell migration. However, in vivo activities of these regulators and how they coordinate to promote efficient cell migration are not understood in detail. The long-term goal is to understand the molecular mechanisms that coordinate directed cell migration. To achieve this goal, the overall objective of this proposal is to determine how Cdc42ep1, an effector protein for Cdc42, interacts with other actin regulators to coordinate neural crest cell migration. Recent study from the lab revealed that Cdc42ep1 is essential in directed migration of neural crest cells during Xenopus embryogenesis. Using this in vivo cell migration system, two subcellular pools of Cdc42ep1 were revealed, one at the protrusive front and the other at the cell body and rear. These two pools of Cdc42ep1 interacts with Cdc42 and septin filaments, respectively, and these interactions can influence the balance of Cdc42ep1 between the two cytoplasmic pools. Therefore, the central hypothesis for this proposal is that Cdc42ep1 coordinates Cdc42- mediated membrane protrusion at the leading edge and the septin-actin cytoskeleton organization at the trailing edge to promote directed neural crest cell migration. The central hypothesis will be tested by pursuing the following three specific aims: 1) Determine the feedback regulation between Cdc42 and Cdc42ep1 and its impact on polarized actin dynamics and directed neural crest cell migration; 2) Determine the mechanisms of cooperation between Cdc42ep1 and septin filaments in controlling cell polarity and directional migration of neural crest cells; and 3) Determine the function and mechanism of septin filaments in regulating the formation and contractility of actin stress fibers. This work is a close collaboration with Tsygankov lab, where a morphodynamic cell migration model will be developed to test various molecular mechanisms and guide further experimental designs. By tightly integrating the in vivo and in silico experiments in a quantitative manner, the proposed research will uncover the mechanisms of how Cdc42ep1 integrates activities of Cdc42 and septin to organize actin dynamics at the protrusive front and the retractive rear to promote neural crest cell migration. This study will fill the knowledge gap of how local cytoskeletal arrangements are coordinated in directed cell migration. This knowledge is not limited to neural crest migration, but can be applied to other migration processes to provide a mechanistic understanding of in vivo cell migration in general. Therefore, the study will not only be critical for understanding the development of neural crest related birth defects, but also help improve our understanding of numerous human diseases that involve dysregulated cell migration in other contexts.

项目概要 细胞迁移对于发育和疾病至关重要，但我们对这种复杂动态的理解 活动相当有限。细胞培养研究发现了肌动蛋白细胞骨架的关键调节因子，包括 GTPase 的 Rho 家族（RhoA、Rac1 和 Cdc42）影响细胞迁移的特定过程。然而， 这些调节因子的体内活性以及它们如何协调以促进有效的细胞迁移尚不清楚 详细了解了。长期目标是了解协调定向的分子机制 细胞迁移。为了实现这一目标，本提案的总体目标是确定 Cdc42ep1（一个 Cdc42 的效应蛋白与其他肌动蛋白调节因子相互作用以协调神经嵴细胞迁移。最近的 实验室研究表明，Cdc42ep1 对于非洲爪蟾期间神经嵴细胞的定向迁移至关重要 胚胎发生。使用这种体内细胞迁移系统，揭示了 Cdc42ep1 的两个亚细胞库，一个是 一个位于突出的前部，另一个位于电池体和后部。这两个 Cdc42ep1 池与 Cdc42 相互作用 和 septin 丝，这些相互作用可以影响 Cdc42ep1 之间的平衡 两个细胞质池。因此，该提案的中心假设是 Cdc42ep1 协调 Cdc42- 介导的前缘膜突出和后缘介导的隔膜肌动蛋白细胞骨架组织 边缘促进神经嵴细胞定向迁移。中心假设将通过追求 以下三个具体目标： 1) 确定 Cdc42 和 Cdc42ep1 之间的反馈调节及其影响 极化肌动蛋白动力学和定向神经嵴细胞迁移； 2）确定机制 Cdc42ep1和septin丝在控制细胞极性和神经定向迁移中的合作 嵴细胞； 3) 确定脓蛋白丝在调节形成和形成中的功能和机制。 肌动蛋白应力纤维的收缩性。这项工作是与 Tsygankov 实验室的密切合作，该实验室的形态动力学 将开发细胞迁移模型来测试各种分子机制并指导进一步的实验 设计。通过以定量方式紧密结合体内和计算机实验，提出了 研究将揭示Cdc42ep1如何整合Cdc42和septin的活性来组织的机制 前突和后缩的肌动蛋白动力学促进神经嵴细胞迁移。这项研究 将填补局部细胞骨架排列在定向细胞迁移中如何协调的知识空白。这 知识不仅限于神经嵴迁移，还可以应用于其他迁移过程，以提供 对体内细胞迁移的一般机制的理解。因此，这项研究不仅对 了解与神经嵴发育相关的先天缺陷，也有助于提高我们对 许多人类疾病都与其他情况下的细胞迁移失调有关。