Mechanisms of mesenchymal chemotaxis

间充质趋化机制

• 批准号: 8890845
• 负责人: JAMES E BEAR
• 金额: $ 35.3万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8890845
• 关键词: Actins; Address; Amplifiers; Architecture; Behavior; Binding; Biochemical; Biological Assay; Blood Platelets; Cardiovascular Diseases; Cardiovascular system; Cells; Chemicals; Chemotaxis; Coagulation Process; Complex; Cues; Cytoskeletal Modeling; Cytoskeleton; Data; Destinations; Disease; Environment; Event; Fibroblasts; Fibrosis; Filopodia; Fingers; Generations; Genetic; Growth Factor; Growth Factor Receptors; Health; Hour; Image; Image Analysis; Lead; Leukocytes; Life; Link; Maintenance; Malignant Neoplasms; Measures; Mechanics; Mediating; Mesenchymal; Mesenchymal Cell Neoplasm; Methods; Microfluidics; Microscopy; Molecular; Motor; Movement; Mutation; Myosin Regulatory Light Chains; Myosin Type II; Nature; Neoplasm Metastasis; Oncogenic; Pathway interactions; Physiological; Physiological Processes; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor Receptor; Process; Publishing; Regulation; Resolution; Role; Shapes; Signal Pathway; Signal Transduction; System; Testing; Therapeutic; Time; Tissues; Total Internal Reflection Fluorescent; Wound Healing; base; cell motility; cell type; cellular imaging; innovation; macrophage; mutant; polymerization; response; spatiotemporal; tumor

DESCRIPTION (provided by applicant): Cells in a variety of contexts migrate towards soluble chemical cues in a process known as chemotaxis. Despite nearly a century of study, the mechanistic underpinnings of chemotaxis remain incompletely understood. Spatial gradients of growth factors direct the movements of mesenchymal cells in tissues to coordinate and accelerate physiologically important processes such as wound healing, and mesenchymal chemotaxis has been implicated in pathological conditions such as cardiovascular and fibrotic diseases. Yet, the vast majority of chemotaxis studies have focused on leukocytes and other fast-moving, amoeboid cells. Mesenchymal chemotaxis has been prohibitively difficult to study, because it requires maintenance of stable gradients for many hours. Traditional methods such as transwell assays provide little or no dynamic information and poorly discriminate effects on the efficiency of motility from actual directional sensing. To overcome these technical limitations we recently established a microfluidic chemotaxis assay that allows direct observation of mesenchymal cells in stable, linear gradients over many hours, allowing both single-cell tracking and high-resolution live-cell imaging approaches such as TIRF microscopy. Our preliminary data indicate that the growth factor receptor, PDGF-R controls mesenchymal chemotaxis by a PLC > PKC > Myosin II pathway and requires the coordination of signaling events and cytoskeletal organization. We propose to elucidate the mechanisms of mesenchymal chemotaxis by 1) Dissecting the spatio- temporal nature of chemotactic signaling in mesenchymal cells 2) Understanding the dynamic organization of the cytoskeleton during chemotaxis in this cell type and 3) Delineating the coordination of signaling and cytoskeletal events that lead to complex chemotactic behaviors such as re-orientation to new cues and chemotaxis in 3D environments. These studies will directly contribute to our understanding the physiological basis of disease states such tumor metastasis, fibrosis and cardiovascular disease, as well as our understanding of physiological processes such as wound healing.

描述（由申请人提供）：在称为趋化性的过程中，各种情况下的细胞向可溶性化学提示迁移。尽管经过近一个世纪的研究，但趋化性的机械基础仍未完全理解。生长因子的空间梯度指导组织中间充质细胞的运动协调和加速生理上重要的过程，例如伤口愈合，间充质趋化性与心血管和纤维化疾病等病理状况有关。然而，绝大多数趋化性研究都集中在白细胞和其他快速移动的变形虫细胞上。间充质趋化性很难研究，因为它需要维持稳定的梯度很多小时。传统方法（例如Transwell Asses）几乎没有或根本没有动态信息，并且与实际方向传感对运动效率的影响很差。为了克服这些技术局限性，我们最近建立了一种微流体趋化性测定法，该测定法可以在许多小时内直接观察稳定的线性梯度中的间充质细胞，从而允许单细胞跟踪和高分辨率的活细胞成像方法，例如TIRF显微镜。我们的初步数据表明，生长因子受体PDGF-R通过PLC> PKC> PKC>肌球蛋白II途径控制间充质趋化性，并且需要信号事件和细胞骨架组织的协调。我们建议通过1）解剖间充质细胞中趋化信号传导的时空性质。 3D环境中的趋化性。这些研究将直接有助于我们理解疾病状态的生理基础，例如肿瘤转移，纤维化和心血管疾病，以及我们对伤口愈合等生理过程的理解。