Cell Signaling and Cell Decisions

细胞信号传导和细胞决策

• 批准号: 10292339
• 负责人: TOBIAS MEYER
• 金额: $ 75.88万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10292339
• 关键词: Actins; Architecture; Biochemical Genetics; Biological Assay; Cell Cycle; Cell Polarity; Cell Proliferation; Cells; Chemotaxis; Complex; DNA; Degenerative Disorder; Development; Disease; Ensure; Event; Family; Feedback; Fluorescence Microscopy; Goals; Human; Immune; Lead; Logic; Maintenance; Malignant Neoplasms; Mammalian Cell; Methods; Mitogens; Modeling; Molecular; Monitor; Monomeric GTP-Binding Proteins; Output; Process; Proliferating; Reporter; Second Messenger Systems; Signal Transduction; Stress; Structure; Therapeutic; Tissues; Work; cell motility; genetic approach; migration; novel therapeutic intervention; polarized cell; prevent; rho; single cell analysis; system architecture; treatment strategy

PROJECT SUMMARY/ABSTRACT Human cells integrate external and internal signaling inputs to make regulatory decisions that change functional outputs. Two such decisions of cells – the decision to polarize and migrate and the decision to enter the proliferative state - are central to multicellular development and tissue maintenance. The regulatory feedbacks and the core switch mechanisms of how cells start to migrate or proliferate are not yet understood. Due to significant cell-to-cell variability and lack of synchronization, an understanding of the underlying regulatory motifs cannot be achieved by biochemical and genetic approaches alone. However, recently developed activity reporters and rapid perturbation strategies have made it possible to investigate complex spatial and temporal signaling feedback architectures and decision processes in living single cells, an approach that can reveal feedback mechanisms and circumvent the technical bulk assay issues. Our work seeks to understand the principles of cellular decision processes in human cells by employing these live-cell methods to monitor, perturb and automatically analyze the relevant signaling processes and ultimately derive quantitative models of how specific decisions are made. Our proposed work has Two Themes: In our first theme, we determine how cells initiate and establish cell polarity and how already polarized cells steer their front during directed migration and chemotaxis. We have developed automated fluorescence microscopy methods to monitor and perturb the critical Rho family small GTPases and relevant second messengers, and developed methods to quantify changes in different actin structures. These approaches will allow us to understand the core regulatory mechanisms for cell polarization and cell steering during migration. In our second theme, we seek to understand how cells decide to transition from a quiescent to a proliferative state by investigating molecular mechanisms of competition between stress and mitogens, by determining the molecular mechanism of the point-of-no return for cell cycle entry, and by exploring how sequential signaling events prevent the re-replication of the same DNA to ensure that DNA is only replicated once. We have developed a number of live cell cycle activity reporters, perturbation strategies and automated single-cell analysis methods that will help us understand and model the regulatory mechanisms controlling human cell cycle entry. Both our themes will lead to new concepts of the logics of human decision processes and will provide detailed molecular and mechanistic models explaining how cells integrate signaling inputs to start to migrate or enter the cell cycle. Finally, the universal dysregulation of cell proliferation and migration in cancer, and the frequent dysregulation of these processes in degenerative, immune and other diseases, argues that a molecular understanding of the complex regulatory architecture of cell migration and proliferation may lead to new therapeutic strategies for the treatment of a broad range of diseases.

项目概要/摘要 人体细胞整合外部和内部信号输入以做出改变的监管决策 细胞的两个这样的决定——极化和迁移的决定以及进入的决定。 增殖状态 - 是多细胞发育和组织维持的核心。 反馈以及细胞如何开始迁移或增殖的核心转换机制尚不清楚。 由于显着的细胞间差异和缺乏同步性，对底层的理解 然而，最近仅通过生化和遗传方法无法实现调节基序。 发达的活动产生和快速扰动策略使得研究复杂的 活单细胞中的空间和时间信号反馈架构和决策过程， 可以揭示反馈机制并规避技术批量分析问题的方法。 试图通过利用这些活细胞来了解人类细胞的细胞决策过程的原理 监控、扰动和自动分析相关信号过程并最终导出的方法 如何做出具体决策的定量模型。 我们提出的工作有两个主题：在第一个主题中，我们确定细胞如何启动和建立 细胞极性以及已经极化的细胞在定向迁移和趋化过程中如何引导其前沿。 开发了自动荧光显微镜方法来监测和干扰关键的 Rho 家族 小 GTP 酶和相关的第二信使，并开发了量化不同变化的方法 这些方法将使我们能够了解细胞的核心调节机制。 在我们的第二个主题中，我们试图了解细胞如何决定。 通过研究竞争的分子机制从静止状态转变为增殖状态 通过确定细胞周期不回归点的分子机制，在应激和有丝分裂原之间进行研究 进入，并通过探索顺序信号事件如何阻止相同 DNA 的重新复制，以确保 DNA只复制一次，我们开发了一些活细胞周期活动产生器，扰动。 策略和自动化单细胞分析方法将帮助我们理解和建模监管 控制人类细胞周期进入的机制。 我们的两个主题都将带来人类决策过程逻辑的新概念，并将提供 详细的分子和机制模型解释细胞如何整合信号输入以开始迁移或 最后，癌症中细胞增殖和迁移的普遍失调，以及 退行性疾病、免疫性疾病和其他疾病中这些过程的频繁失调，认为 对细胞迁移和增殖的复杂调控结构的分子理解可能会导致 治疗多种疾病的新治疗策略。