Functional causality in regulating cell morphogenesis

调节细胞形态发生的功能因果关系

• 批准号: 10401805
• 负责人: Gaudenz Danuser
• 金额: $ 85.9万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10401805
• 关键词: 3-Dimensional; Actins; Address; Adopted; Algorithms; Architecture; Cell Shape; Cell physiology; Cells; Cellular biology; Chemicals; Complex; Computer Vision Systems; Coupling; Cytoskeleton; Development; Disease; Environment; Etiology; Experimental Designs; Feedback; Laboratories; Lead; Mechanics; Mediating; Molecular; Morphogenesis; Pathology; Pathway interactions; Phenotype; Physiology; Polymers; Process; Regulation; Research; Research Methodology; Series; Shapes; Signal Transduction; Statistical Models; System; Time; Tissues; Work; base; imaging modality; innovation; insight; live cell imaging; live cell microscopy; mathematical model; mechanical force; programs; response; tool

Morphogenesis is a fundamental process in a wide range of cell functions and therefore one of the best studied. Consequently, most of the contributing component processes and the majority of their molecular parts are known. Much less understood is how the many underlying component processes are integrated into a working whole. This integration is regulated by a system of chemical and mechanical pathways with i) a high level of non-linearity; ii) a high level of redundancy; and iii) a separation in space and time of cause and effect between component processes. A key consequence of such a pathway configuration is that perturbation of any component can lead to wide-ranging and fast adaptation. Hence, phenotypic changes primarily reflect a reconfiguration of the system and not necessarily the function of the perturbed target. This challenge has plagued the interrogation of molecular functions in cell morphogenesis, and has led to numerous controversies that likely relate to slight differences in experimental designs triggering divergent adaptation processes. To circumvent some of these limitations, my lab has developed over the past 10 years quantitative live cell imaging methods that reveal the functional interplay between spatially and temporally distributed molecular processes, such as cytoskeleton polymer dynamics, forces, and chemical signals, based on the coupling of their spontaneous activation fluctuations in unperturbed systems. Building on key discoveries of cell morphogenic control mechanisms, which required the use of a perturbation-free approach rather than a more conventional experimental paradigm, we propose here an extension of this research program that will introduce a rigorous statistical framework to infer the coupling of molecular processes in cause and effect cascades. This includes algorithms to identify directly, from fluctuation time series, feedback interactions between processes and the dynamic rewiring of the cause and effect cascades under variable cellular conditions. These computational developments will be paralleled by innovation in experimental systems for multispectral live cell imaging of up to 8 concurrent processes and for the analysis of cell morphogenesis in native tissue environments in 3D. The studies will be focused on the coordination of the system of highly redundant actin modulating factors in promoting actin mediated cell shape changes and the interactions of this system with the regulatory system of RhoGTPase signals. The impact of this work will reach far beyond the new insights we will gain of the regulation of these pathway systems. The work will address the notorious adaptation responses of complex molecular systems, which are generic and a fundamental impediment to the systematic inquiry of cell functions. The tools we develop to overcome some of these problems will be made available as widely-adoptable approaches to the analysis of cell regulatory processes.

形态发生是一个基本的过程，在广泛的细胞功能，因此最好的一个 研究了因此，大多数起作用的组分过程和它们的大部分分子部分 是已知的。更少有人了解的是，许多底层组件流程如何集成到一个 整体工作。这种整合是由化学和机械途径系统调节的， 非线性水平; ii）高冗余水平;以及iii）因果关系在空间和时间上的分离 组件进程之间。这样的路径配置的关键结果是， 组件可以导致广泛和快速的适应。因此，表型变化主要反映了 系统的重新配置，而不一定是扰动目标的功能。这一挑战 困扰着细胞形态发生中的分子功能的询问，并导致了许多争议 这可能与引发不同适应过程的实验设计的细微差异有关。到 克服了这些限制，我的实验室在过去10年里开发了定量活细胞 显示空间和时间分布的分子之间的功能相互作用的成像方法 过程，如细胞骨架聚合物动力学，力，和化学信号，基于耦合， 它们在未扰动系统中的自发激活波动。基于细胞的关键发现 形态发生的控制机制，这需要使用一个无扰动的方法，而不是一个更 传统的实验范式，我们在这里提出了这个研究计划的扩展，将 引入严格的统计框架来推断因果关系中分子过程的耦合 瀑布这包括从波动时间序列中直接识别反馈相互作用的算法 在不同的细胞状态下， 条件这些计算的发展将被实验系统的创新所取代， 多光谱活细胞成像多达8个并发过程，并用于分析细胞形态发生， 3D中的自然组织环境。这些研究将集中在系统的协调高度 多余的肌动蛋白调节因子在促进肌动蛋白介导的细胞形状变化中的作用以及这种相互作用 系统与RhoGT 3信号的调节系统。这项工作的影响将远远超出 新的见解，我们将获得这些途径系统的调节。这项工作将解决臭名昭著的 复杂分子系统的适应反应，这是通用的，是一个根本性的障碍， 细胞功能的系统研究。我们开发的克服这些问题的工具将被用于 可作为广泛采用的方法来分析细胞调节过程。