Feedback and Crosstalk in Eukaryotic Chemotaxis

真核趋化中的反馈和串扰

• 批准号: 8109302
• 负责人: Takanari Inoue
• 金额: $ 30.85万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8109302
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute; Address; Arthritis; Automobile Driving; Biological Process; CCI-779; Calcium Signaling; Cell Cycle; Cell physiology; Cells; Cellular biology; Characteristics; Chemistry; Chemotactic Factors; Chemotaxis; Complication; Development; Disease; Disease Progression; Dissection; Embryonic Development; Engineering; Event; Feedback; Functional disorder; Generations; Genetic; Goals; Image; Imaging Techniques; Immune; Impairment; Kinetics; Knowledge; Life; Malignant Neoplasms; Metabolic; Methodology; Methods; Microfluidic Microchips; Modeling; Molecular; Monitor; Morphology; Natural regeneration; Neurons; PTEN gene; Pathway interactions; Pattern; Pharmacology; Physiological; Process; Proteins; Public Health; RNA Interference; Reaction; Research; Resolution; Signal Transduction; Signaling Molecule; Specificity; System; Techniques; Testing; Therapeutic; Time; Tissues; Wound Healing; angiogenesis; biological systems; cell motility; fluorescence imaging; improved; interest; migration; molecular dynamics; neutrophil; novel; operation; public health relevance; receptor; rho GTP-Binding Proteins; tool

DESCRIPTION (provided by applicant): Chemotaxis occurs during a number of physiological events including angiogenesis, embryonic development, wound healing, immune defense, and the establishment of neuronal circuits. Accordingly, eukaryotic chemotaxis has been a topic of key interest in cell biology and pathophysiology. The objective of our research is to explore a long-standing conundrum in the field: How do cells "break symmetry" to initiate cell migration? To address this question, we will focus on two signaling modules that remain uncharacterized: "Feedback" and "Crosstalk". Experimental dissection of these modules has proven inherently challenging, because their signaling operation in cells is local (sub-cellular) and rapid (second-to-minute) on top of their intimate intra- and inter-module relationships. The experimental perturbation of component molecules thus must be restricted to precise spatial domains and be faster than the signaling events. However, most tools used to probe signaling events are generally slow (minute- to-day) and global (supra-cellular) in their effects, limiting their usefulness. We previously developed a new generation of molecular tools that allows for Rapid, Inducible and Specific Perturbation (RISP) of various proteins in living cells. In order to decipher the kinetics and dynamics of molecular networks within the modules with high spatio-temporal resolution, we employ two different approaches: 1. operating the RISP in microfluidic device and 2. improving the present RISP by using synthetic photo- chemistry knowledge. These experimentations will allow us to determine whether and how the elementary signaling modules are integrated to orchestrate an intricate symmetry breaking process. A better understanding of the chemotaxis promises therapeutic advancements for cell migration-related diseases. Our unique approach for probing cellular dynamics will also provide a general-and-powerful methodology that has the potential to significantly extend conventional methods such as RNA interference and pharmacology. PUBLIC HEALTH RELEVANCE: Due to its significant involvement in biological processes, cell migration contributes to disease progression in cancer and arthritis, while its impairment leads to anomalous tissue development or regeneration. Our unique perturbation approach can be a powerful strategy for not only dissecting the molecular mechanisms, but also interfering with these cell migration-related diseases.

描述（由申请人提供）：趋化性发生在许多生理事件中，包括血管生成，胚胎发育，伤口愈合，免疫防御和神经元回路的建立。因此，真核趋化性一直是细胞生物学和病理生理学关键兴趣的话题。我们研究的目的是探索该领域的长期难题：细胞如何“打破对称性”以启动细胞迁移？为了解决这个问题，我们将重点关注两个保持未表征的信号模块：“反馈”和“串扰”。这些模块的实验解剖已经证明了固有的具有挑战性，因为它们在细胞中的信号传导操作是局部（次细胞），并且在其亲密的内部和模块间关系的顶部是局部（二次到二分钟）。因此，组分分子的实验扰动必须限制为精确的空间结构域，并且比信号事件更快。但是，大多数用于探测信号事件的工具通常是缓慢的（今天的分钟）和全局（超细胞）的效果，从而限制了它们的实用性。我们以前开发了一种新一代的分子工具，该工具允许活细胞中各种蛋白质的快速，诱导和特定的扰动（RISP）。为了解读具有高时空分辨率的模块内分子网络的动力学和动力学，我们采用了两种不同的方法：1。在微流体设备中操作RISP和2。通过使用合成化学照片知识来改善当前的RISP。这些实验将使我们能够确定是否以及如何整合基本信号模块以协调复杂的对称破坏过程。对趋化性的更好理解有望针对细胞迁移相关疾病的治疗进步。我们探测细胞动力学的独特方法还将提供一种通用和功能的方法，该方法有可能显着扩展常规方法，例如RNA干扰和药理学。 公共卫生相关性：由于其在生物过程中的大量参与，细胞迁移有助于癌症和关节炎的疾病进展，而其损害会导致异常的组织发育或再生。我们独特的扰动方法不仅可以剖析分子机制，还可以干扰这些与细胞迁移相关的疾病。