Symmetry breaking and polarization of cell in 3D environments

3D 环境中细胞的对称性破缺和极化

• 批准号: 9049973
• 负责人: Kevin Michael Dean
• 金额: $ 5.88万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9049973
• 关键词: Actins; Actomyosin; Acute; Address; Adhesions; Adopted; Affect; Animal Model; Anisotropy; Apoptosis; Architecture; Biochemical; Biological; Biological Process; Biosensor; Bundling; Cell Differentiation process; Cell Polarity; Cell Shape; Cell membrane; Cell physiology; Cell surface; Cells; Cellular Morphology; Cellular biology; Characteristics; Collagen; Collagen Fiber; Complex; Computer Vision Systems; Coupling; Cues; Cytoskeleton; Data; Disease; Embryonic Development; Environment; Event; Evolution; Extracellular Matrix; F-Actin; Family; Feedback; Fiber; Fluorescence; Fluorescence Resonance Energy Transfer; Focal Adhesions; Foundations; Future; Guanosine Triphosphate Phosphohydrolases; Health; Human; Image; Imaging technology; Integrins; Light; Maintenance; Maps; Mechanics; Mediating; Mesenchymal; Microfilaments; Microscope; Microscopy; Molecular; Neoplasm Metastasis; Outcome; Physiological; Postdoctoral Fellow; Primary Neoplasm; Process; Psychological reinforcement; Radial; Reporting; Research; Resistance; Resolution; Scientist; Signal Transduction; Site; Staging; Structure; Testing; Tissues; Training; Work; cell motility; clinically significant; depolymerization; extracellular; insight; live cell imaging; migration; novel; polarized cell; polymerization; public health relevance; reconstitution; response; rho; single molecule; spatiotemporal; three dimensional structure; three-dimensional modeling

 DESCRIPTION (provided by applicant): The mechanics, 3D structure, and biochemical composition of the extracellular matrix (ECM) significantly influences diverse biological outcomes, including cellular polarization, differentiation, apoptosis, migration, and proliferation For example, ECM alignment triggers polarized cell morphologies and facilitates migration away from the primary tumor. However, it remains poorly understood how cells spatiotemporally integrate 3D ECM cues in order to selectively orchestrate downstream signaling. The objective of the proposed research here aims to further our understanding this process. Specifically, I will investigate how mesenchymal cells spontaneously establish anteroposterior cell polarity in 3D ECM environments. Towards these means, I will evaluate the spatiotemporal evolution of cell-collagen interfaces, integrin activation, Rho family GTPase signaling, and cytoskeletal dynamics, in mesenchymal cells undergoing 3D polarization. I hypothesize that ECM fibers oriented normal to the cell surface will trigger integrin clustering and the formation of mature matrix adhesions (Aim 1). Furthermore, I hypothesize that Rho family GTPase activation will be bifurcated into focal adhesion-dependent and independent regimes (Aim 2), and that this will result in ECM context-dependent modulation of actin polymerization, bundling, dendritic branching, actomyosin contraction, and depolymerization (Aim 3). This will be accomplished by light- sheet imaging of mesenchymal cells embedded within reconstituted ECM-like environments, acute pharmacological perturbations, and computer vision analysis of ECM fiber orientation, cell shape, signal transduction, and cytoskeletal dynamics. This research will serve as the foundation from which further studies can evaluate 3D spatiotemporal cell signaling in physiological and pathophysiological ECM states.

 描述（由申请人提供）：细胞外基质（ECM）的力学、3D结构和生化成分显着影响多种生物学结果，包括细胞极化、分化、凋亡、迁移和增殖。例如，ECM排列触发极化的细胞形态并促进远离原发肿瘤的迁移。然而，人们对细胞如何时空整合 3D ECM 线索以选择性地协调下游信号传导仍知之甚少。这里提出的研究的目的是为了进一步我们理解这个过程。具体来说，我将研究间充质细胞如何在 3D ECM 环境中自发建立前后细胞极性。针对这些方法，我将评估经历 3D 极化的间充质细胞中细胞-胶原蛋白界面、整合素激活、Rho 家族 GTPase 信号传导和细胞骨架动力学的时空演化。我假设 ECM 纤维垂直于细胞表面定向将触发整合素聚集和成熟基质粘附的形成（目标 1）。此外，我假设 Rho 家族 GTPase 激活将分为粘着斑依赖性和独立状态（目标 2），这将导致肌动蛋白聚合、成束、树突分支、肌动球蛋白收缩和解聚的 ECM 环境依赖性调节（目标 3）。这将通过嵌入重建的 ECM 样环境中的间充质细胞的光片成像、急性药理学扰动以及 ECM 纤维取向、细胞形状、信号转导和细胞骨架动力学的计算机视觉分析来实现。这项研究将为进一步研究评估生理和病理生理 ECM 状态下的 3D 时空细胞信号传导奠定基础。