Feedback and Crosstalk in Eukaryotic Chemotaxis- Administrative Supplement

真核趋化性中的反馈和串扰-行政补充

• 批准号: 8703909
• 负责人: Takanari Inoue
• 金额: $ 3.92万
• 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 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8703909
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute; Address; Administrative Supplement; 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; Event; Feedback; Functional disorder; Generations; Genetic; Genetic Engineering; 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; 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; 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.

描述（由申请人提供）：趋化性发生在许多生理事件期间，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的一个重要课题。我们研究的目的是探索该领域长期存在的难题：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尚未表征的信号模块：“反馈”和“串扰”。事实证明，这些模块的实验剖析本质上具有挑战性，因为它们在细胞中的信号传导操作是局部的（亚细胞的）并且是快速的（秒到分钟的），而且除了模块内和模块间的密切关系之外。因此，成分分子的实验扰动必须限制在精确的空间域内，并且比信号事件更快。然而，大多数用于探测信号事件的工具的效果通常很慢（每分钟）和全局（超细胞），限制了它们的实用性。我们之前开发了新一代分子工具，可以对活细胞中的各种蛋白质进行快速、诱导和特异性扰动 (RISP)。为了以高时空分辨率破译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.在微流体装置中操作RISP；2.利用合成光化学知识改进现有的RISP。这些实验将使我们能够确定是否以及如何集成基本信号模块来协调复杂的对称性破坏过程。对趋化性的更好理解有望在细胞迁移相关疾病的治疗方面取得进展。我们探测细胞动力学的独特方法还将提供一种通用而强大的方法，有可能显着扩展 RNA 干扰和药理学等传统方法。