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: 10197977
• 负责人: Bo Sun
• 金额: $ 35.25万
• 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/10197977
• 关键词: 3-Dimensional; Anisotropy; Area; Cells; Chemicals; Collagen; Computer Models; Crosslinker; Cues; DNA; DNA Sequence; Development; Dimensions; Disease Progression; Engineering; Environment; Epithelial; Exhibits; Extracellular Matrix; Foundations; Goals; Human Biology; Image; Immune System Diseases; Knowledge; Label; Lead; Life; Light; Malignant Neoplasms; Markov Chains; Measures; Mechanics; Mesenchymal; Molecular; Neoplasm Metastasis; Organoids; Pattern; Periodicity; Physics; Physiological Processes; Regulation; Research; Role; Stimulus; Stress; Techniques; Testing; Therapeutic; Time; Tissue Engineering; Tissues; base; cancer therapy; cell motility; cell type; deep learning; design; digital; fascinate; genetically modified cells; in vivo; migration; nanoparticle; programs; protein protein interaction; response; synergism; tissue regeneration; transcription factor

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 mode transitions of cells in 3D extracellular matrix (ECM). My lab has developed deep-learning based image postprocessing to track the migration modes of cells. We also developed techniques to manipulate and measure the micromechanics of ECM at cellular scale. Based on these preliminary results, I will systematically study the intrinsic and extrinsic control mechanisms of 3D cell migration mode transitions in collagen ECM. The results will pave the way for my long-term goals to understand the organizing principle that lead molecules to life, and to program cell motility for applications in tissue engineering and cancer treatment. To this end, I will dedicate my lab to the following research thrusts. Thrust 1 aims to determine how cell migration mode transitions are regulated by external cues, as well as intrinsic states of cells during the Epithelial-Mesenchymal Transition (EMT). I will test three hypotheses that elucidate the roles of ECM micromechanical stiffness, anisotropy, plasticity, synergy of mechanical and chemical guidance, as well as EMT stage in modulating the cell migration mode transitions. I will employ sophisticated ECM engineering and characterization techniques developed in my lab. I will also use genetically engineered cells whose EMT transcription factors are fluorescent labeled and can be specifically activated. Completion of thrust 1 will establish 3D cell migration as a hidden Markov process where the mesoscale dynamics, namely the migration mode transitions, provides a unifying framework to explain diverse dynamic patterns of 3D cell migration observed in vivo. Thrust 2 aims to devise strategies to program cell migration via nonstationary mechanical cues. In subproject 1, I will employ techniques developed in my lab to control 3D contact guidance cues in space and in real time. By measuring the migration mode transitions under step-increasing contact guidance, I will obtain the energy barriers that separate different modes. Then under periodic mechanical stimuli I will measure and computationally model the nonequilibrium mode transition flux, a statistical physics quantity that inform the efficiency and energy dissipation of cell motility responses. These mesoscale quantities shed light to the underlying molecular organizing principles. In subproject 2 I will develop collagen ECM which exhibits digital response to stresses using DNA-grafted nanoparticles as crosslinkers. I will design the DNA sequence to control the yield strength of crosslinkers, thereby programing cell migration mode both for single cell and for collective organoid migration. Completion of thrust 2 will expands the design space of engineered ECM, laying a foundation for the mechanical programing of 3D cell motility.

3D组织空间中的细胞迁移对于人类生物学至关重要。但是，预测和 尽管有牢固的分子机器，但编程3D细胞运动仍然是主要挑战 涉及。填补压倒性亚细胞细节（例如蛋白质 - 蛋白质）之间的知识差距 相互作用以及组织空间中不同细胞类型表现出的引人入胜的动态模式，我将集中精力 在中尺度细胞动力学上，即3D细胞外基质中细胞的迁移模式转变 （ECM）。我的实验室已经开发了基于深度学习的图像后处理，以跟踪细胞的迁移模式。 我们还开发了在细胞尺度下操纵和测量ECM的微力学的技术。基于 在这些初步结果上，我将系统地研究3D细胞的内在和外在控制机制 胶原蛋白ECM中的迁移模式过渡。结果将为我的长期目标铺平道路 使分子栩栩如生的组织原理，并为在组织中应用编程细胞运动 工程和癌症治疗。为此，我将把实验室奉献给以下研究推力。推力1 旨在确定细胞迁移模式过渡如何受外部线索以及内在状态调节 上皮间质转变（EMT）期间的细胞。我将检验三个阐明角色的假设 ECM微机械刚度，各向异性，可塑性，机械和化学指导的协同作用 作为调节细胞迁移模式过渡的EMT阶段。我将采用精致的ECM工程 和在我的实验室中开发的表征技术。我还将使用EMT的基因工程细胞 转录因子是荧光标记的，可以特异性激活。推力1的完成将建立 3D细胞迁移是一个隐藏的马尔可夫过程，其中中尺度动力学，即迁移模式 过渡提供了一个统一的框架，以解释在3D细胞迁移中观察到的各种动态模式 体内。推力2旨在制定通过非组织机械提示编程细胞迁移的策略。在 第1个子标记1，我将在实验室中采用开发的技术来控制太空中的3D接触指导线索 即时的。通过测量逐步增加的接触指导下的迁移模式过渡，我将获得 分开不同模式的能屏障。然后在周期性的机械刺激下，我将测量和 计算对非平衡模式过渡通量建模，这是一种统计物理数量，告知 细胞运动反应的效率和能量耗散。这些中尺度的数量使光明 基础分子组织原理。在第2台中，我将开发胶原蛋白ECM，该胶原ECM展示数字 对应力的反应，使用DNA接枝的纳米颗粒作为交联。我将设计DNA序列以控制 交联器的屈服强度，从而为单个单元和集体编程细胞迁移模式 器官迁移。推力2的完成将扩大工程ECM的设计空间，奠定基础 用于3D细胞运动的机械编程。