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

细胞在三维组织空间中的迁移对人类生物学具有重要意义。然而，预测和 编程3D细胞运动仍然是主要的挑战，尽管对分子机械的了解很清楚 牵涉其中。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，以及不同类型的细胞在组织空间中展示的迷人的动态模式，我将重点放在 关于中尺度细胞动力学，即细胞在三维细胞外基质中的迁移表型转变 (ECM)。为了推进家长奖的目标，将在家长奖的范围内追求两个具体目标 获奖。这些特定的目标依赖于使用先进的专用荧光显微镜 光电操控模块。特别是，Aim 1将阐明迁移表型是如何受 ECM机械线索的时空梯度。我将测量迁徙表型转换率 编码细胞对ECM力学时空梯度的响应。电解加工的力学性能 将通过利用光活化的ECM交联剂进行实时控制。这一结果将导致 构建了定量刻画细胞迁移表型可塑性的准势能景观。 在目标2中，我将使用可光转换的荧光细胞标记来确定迁移表型特征 部分上皮-间充质转化(EMT)状态。我会将迁徙的表型景观与 用转化生长因子-β处理同一上皮球的上皮细胞、间充质细胞和部分内胚层细胞。我也会 用潜在的基因表达网络证实了这一发现，因此整合了上游和 下游观察以建立解释细胞迁移的启动和演化的机制模型 模式可塑性。这两个目标都与父奖项一致，同时提高了数据质量和进一步的机械化 洞察力。为了实现这些目标，我将购买一台配备最先进成像和 光电操控模块。该系统将取代老化了8年的荧光显微镜，其中之一 圆周率的主要乐器。获得的设备将全额分配给母公司奖励支持的项目 并对这一应用进行了研究。

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.