Understanding the control mechanisms of 3D cell migration from new dimensions

从新维度理解3D细胞迁移的控制机制

• 批准号: 10579538
• 负责人: Bo Sun
• 金额: $ 16.7万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579538
• 关键词: 3-Dimensional; 8 year old; Aging; Area; Award; Cells; Development; Dimensions; Disease Progression; Environment; Epithelial; Equipment; Evolution; Exhibits; Extracellular Matrix; Gene Expression; Goals; Human Biology; Image; Immune System Diseases; Knowledge; Lead; Malignant Neoplasms; Measures; Mechanics; Mesenchymal; Modeling; Molecular; Neoplasm Metastasis; Parents; Pattern; Phenotype; Physiological Processes; Regulation; Research; System; Therapeutic; Time; Tissues; Transforming Growth Factor beta; Transitional Cell; base; cell motility; cell type; crosslink; data quality; fascinate; fluorescence microscope; improved; insight; instrument; mechanical properties; mechanical signal; migration; programs; protein protein interaction; response; tissue regeneration

Cell migration in 3D tissue space is of fundamental importance for human biology. However, predicting and programming 3D cell motility remain as major challenges despite of a firm picture of the molecular machineries involved. To fill the knowledge gap between the overwhelming subcellular details such as protein-protein interactions, and the fascinating dynamic patterns exhibited by different cell types in tissue spaces, I will focus on the mesoscale cellular dynamics, namely the migration phenotype transitions of cells in 3D extracellular matrix (ECM). To advance the goal of the parent award, two specific aims will be pursued within the scope of the parent award. These specific aims rely on access to a dedicated fluorescent microscope with advanced photomanipulation modules. In particular, aim 1 will elucidate how migration phenotype is modulated by the spatial-temporal gradient of ECM mechanical cues. I will measure the migration phenotype transition rates which encode the cellular responses to spatial-temporal gradient of ECM mechanics. The ECM mechanical properties will be controlled in real time by taking advantage of photoactivated ECM crosslinks. The result will lead to the construction of quasipotential energy landscape that quantitatively depict the cell migration phenotype plasticity. In aim 2 I will employ photo convertible fluorescent cell markers to determine the migration phenotype signatures of partial Epithelial-Mesenchymal Transition (EMT) states. I will compare the migration phenotype landscape for epithelial, mesenchymal, and partial EMT cells from the same epithelial spheroid treated by TGF-β. I will also corroborate the findings with underlying gene expression networks, therefore integrating upstream and downstream observations to build mechanistic models that explain the initiation and evolution of cell migration mode plasticity. Both aims are inline with the parent award, while improving data quality and further mechanistic insights. To pursue the aims I will purchase a fluorescent microscope equipped with state-of-the-art imaging and photomanipulation modules. This system will replace an aging 8 years old fluorescent microscope, one of the PI’s main instrument. The acquired equipment will be fully allocated to the project supported by the parent award and the research in this application.

3D 组织空间中的细胞迁移对于人类生物学至关重要。然而，预测和 尽管对分子机器有了清晰的认识，但对 3D 细胞运动进行编程仍然是主要挑战 涉及。填补蛋白质-蛋白质等压倒性亚细胞细节之间的知识空白 相互作用，以及组织空间中不同细胞类型表现出的迷人动态模式，我将重点关注 中尺度细胞动力学，即细胞在 3D 细胞外基质中的迁移表型转变 （ECM）。为了推进家长奖的目标，将在家长范围内追求两个具体目标 奖。这些具体目标依赖于使用具有先进功能的专用荧光显微镜 光操纵模块。特别是，目标 1 将阐明迁移表型是如何受 ECM 机械线索的时空梯度。我将测量迁移表型转变率 编码细胞对 ECM 力学时空梯度的反应。 ECM机械性能 将通过利用光激活 ECM 交联进行实时控制。结果将导致 构建定量描述细胞迁移表型可塑性的准势能景观。 在目标 2 中，我将采用光可转换荧光细胞标记来确定迁移表型特征 部分上皮-间质转化（EMT）状态。我将比较迁移表型景观 来自经 TGF-β 处理的同一上皮球体的上皮细胞、间充质细胞和部分 EMT 细胞。我也会 用潜在的基因表达网络证实这些发现，从而整合上游和 下游观察以建立解释细胞迁移的起始和演化的机制模型 模式可塑性。这两个目标都与家长奖一致，同时提高数据质量和进一步的机制 见解。为了实现这一目标，我将购买一台配备最先进成像技术的荧光显微镜 光操纵模块。该系统将取代已经使用 8 年的荧光显微镜，该显微镜是 PI的主要仪器。收购的设备将全部分配给母公司资助的项目 以及本应用的研究。