Multi-cue Guidance of Mesenchymal Cell Migration

间充质细胞迁移的多线索引导

• 批准号: 10552599
• 负责人: Jason M. Haugh
• 金额: $ 28.88万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552599
• 关键词: Acceleration; Actins; Affect; Biological Assay; Cardiovascular Diseases; Cardiovascular system; Cells; Cellular biology; Chemicals; Chemotactic Factors; Chemotaxis; Complex; Cues; Cytoskeleton; Disease; Disseminated Malignant Neoplasm; Elements; Engineering; Extracellular Matrix; F-Actin; Feedback; Fibroblasts; Fibrosis; Generations; Image Analysis; Immobilization; Integrins; Invaded; Leukocytes; Ligands; Location; Mediating; Mesenchymal; Microfluidics; Microscopy; Molecular; Movement; Myosin Regulatory Light Chains; Neoplasm Metastasis; Nonmuscle Myosin Type IIA; Pathologic; Pathway interactions; Phospholipase C; Phosphorylation; Physiological; Physiological Processes; Platelet-Derived Growth Factor; Play; Polymers; Process; Productivity; Protein Kinase C; Regulation; Regulatory Pathway; Role; Shapes; Signal Pathway; Signal Transduction; Signaling Protein; Surface; Testing; Time; Tissues; Variant; Work; cell behavior; cell motility; combinatorial; density; in vivo; interdisciplinary approach; link protein; migration; non-muscle myosin; polymerization; synergism; tumor; wound; wound healing

SUMMARY 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 platelet-derived growth factor (PDGF) and other chemoattractants 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. Despite the central role that fibroblasts and other mesenchymal cells play in wound healing and other disease processes such as metastatic cancer and fibrosis, a rigorous understanding of mechanisms governing the directed migration of mesenchymal cells is only recently emerging. To advance further, a quantitative, integrative approach is required. Specifically, it is necessary to elucidate how the central regulatory pathways network with others and how they are coordinated with respect to subcellular location and time to affect cell behavior. In the context of directed mesenchymal cell migration, another layer of complexity is the variation of gradient conditions (midpoint concentration/surface density and steepness). Enabled by new engineering advances, we are poised to tackle these new questions related to chemotaxis and haptotaxis and to their combinatorial influence in multi-cue settings. Our Specific Aims are as follows: Aim 1: Decoding the dynamics of multiple signaling axes that shape mesenchymal chemotaxis. We will test the hypothesis that protrusion dynamics are governed by the metastable push/pull of Arp2/3 complex and NMII activities, which are insufficiently biased by a chemotactic gradient. With stable polarization of active PKC in the most-up-gradient protrusion, the inactivation of NMII there provides a ‘port in the storm’ for pro-Arp2/3 signaling to mediate more productive protrusion. Aim 2: Probing the dynamics of haptotactic sensing and signal amplification. We hypothesize that differential integrin engagement on ECM gradients drives significant cell migration bias through feedback amplification of the pro-Arp2/3 signaling axis. If so, it would imply that haptotactic gradients are able to bias pro-Arp2/3 signaling in mesenchymal cells to an extent that chemotactic gradients cannot. Aim 3: Defining gradient synergy and prioritization in multi-cue scenarios. Despite the relevance for guidance of mesenchymal cells in vivo, it is completely unknown how cells respond to co-presentation of the two gradient types in a controlled setting. Considering how chemotaxis and haptotaxis affect dynamic regulation of the actin cytoskeleton in fibroblasts, we hypothesize that the two gradients synergize when presented in a parallel orientation. By presenting the gradients in an antiparallel or orthogonal orientation, we will determine how cells prioritize the two types of cues.

总结 细胞在各种环境中迁移到可溶性化学线索的过程称为趋化性。 尽管近世纪的研究，趋化性的机械基础仍然不完全 明白血小板衍生生长因子（PDGF）和其他化学引诱物的空间梯度直接 间充质细胞在组织中的运动，以协调和加速生理上重要的 过程，如伤口愈合，和间充质趋化性已牵连到病理 例如心血管和纤维化疾病。尽管成纤维细胞和其他细胞在这一过程中发挥着重要作用， 间充质细胞在伤口愈合和其他疾病过程中起作用，例如转移性癌症， 纤维化，对间充质细胞定向迁移机制的严格理解 才刚刚出现为了进一步取得进展，需要采取定量的综合办法。 具体而言，有必要阐明中枢调控通路如何与其他通路联网，以及如何与其他通路联网。 它们相对于亚细胞位置和时间进行协调以影响细胞行为。背景下 定向间充质细胞迁移，另一层复杂性是梯度条件的变化 （中点浓度/表面密度和陡度）。通过新的工程技术进步，我们 准备解决这些与趋化性和趋触性以及它们的组合有关的新问题， 在多线索设置的影响。我们的具体目标如下： 目的1：解码形成间充质趋化性的多个信号轴的动力学。 我们将检验这一假设，即前突动力学是由Arp 2/3的亚稳态推/拉控制的 复合物和NMII活性，其不足以通过趋化梯度偏向。稳定 在最高梯度突起的活性PKC的极化中，那里的NMII的失活提供了 “风暴中的港口”，用于pro-Arp 2/3信号传导以介导更有生产力的突出。 目的二：探讨触觉感受和信号放大的动力学。我们假设 ECM梯度上的差异整合素接合通过反馈驱动显著的细胞迁移偏倚 pro-Arp 2/3信号轴的扩增。如果是这样的话，这将意味着触觉梯度能够 使间充质细胞中的pro-Arp 2/3信号传导偏向到趋化梯度不能达到的程度。 目标3：确定多线索情景中的梯度协同作用和优先次序。尽管相关性 对于体内间充质细胞的引导，细胞如何响应共呈递是完全未知的。 在受控设置中的两种梯度类型。考虑到趋化性和趋触性如何影响 动态调节的肌动蛋白细胞骨架在成纤维细胞，我们假设，这两个梯度 当以平行方向呈现时协同作用。通过以反平行或 正交方向，我们将确定细胞如何优先考虑这两种类型的线索。