Feedback and Crosstalk in Eukaryotic Chemotaxis

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，真核细胞的趋化性一直是细胞生物学和病理生理学中的一个重要课题。我们的研究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，我们将集中在两个信号模块，仍然没有特征：“反馈”和“串扰”。这些模块的实验解剖已被证明具有内在的挑战性，因为它们在细胞中的信号传导操作是局部的（亚细胞）和快速的（秒到分钟），在它们的亲密内部和模块间的关系之上。因此，组分分子的实验扰动必须限制在精确的空间域，并且比信号事件更快。然而，大多数用于探测信号传导事件的工具在其作用方面通常是缓慢的（分钟到天）和全局的（超细胞的），限制了它们的有用性。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。在微流控装置中操作RISP，以及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基本信号模块是否以及如何整合以协调复杂的对称性破缺过程。更好地了解趋化性有望为细胞迁移相关疾病的治疗进展。我们探测细胞动力学的独特方法也将提供一种通用和强大的方法，有可能显着扩展传统的方法，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重要参与，细胞迁移有助于癌症和关节炎的疾病进展，而其损伤导致异常的组织发育或再生。我们独特的微扰方法可以成为一种强大的策略，不仅可以剖析分子机制，还可以干扰这些细胞迁移相关疾病。

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.