Mechanisms and Functions of Unconventional Intercellular Calcium Waves in Electrically Non-excitable Cells

电不可兴奋细胞中非常规细胞间钙波的机制和功能

• 批准号: 10714066
• 负责人: Xin Tang
• 金额: $ 36.26万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714066
• 关键词: 3-Dimensional; Actomyosin; Algorithms; Biological Process; CRISPR imaging; Calcium; Calcium Oscillations; Calcium Signaling; Cell Line; Cell physiology; Cells; Chemicals; Dissection; Epithelial Cells; Foundations; Gene Expression; Genetic; Grant; Heart; Knowledge; Link; Mechanics; Mediating; Molecular; Organism; Pathology; Physiological; Process; Proteins; Research; Signal Transduction; System; Therapeutic; Work; cell behavior; cell motility; extracellular; innovation; insight; live cell imaging; mechanical stimulus; mechanotransduction; novel; programs; sensor; success; transcriptome

Project Summary Mechanotransduction is the process by which cells sense and transduce extracellular mechanical stimuli into intracellular signaling and gene expression. Mechanotransduction is ubiquitous across diverse organisms and has significant influences on cell function and behavior, in parallel with chemical and genetic signal transductions. The mechanics of microenvironments mediate mechanotransduction through cell-microenvironment interactions and its mis-regulation is at the heart of various pathologies. A major knowledge gap in the field is how mechanical stimuli from microenvironments are transduced into cellular signaling and what the relationships are between microenvironmental mechanics, cell signaling, gene expression, and cell functions. We recently discovered that multiple electrically non-excitable epithelial cell lines can initiate and propagate spontaneous long-distance intercellular calcium waves (ICWs), when cells are cultured in 2D/3D soft microenvironments, but not in stiff ones. Because calcium regulates a broad range of essential cell functions, our findings uncover an unprecedented mechanotransduction nexus between microenvironmental mechanics and diverse cellular signals. We hypothesize that the cellular actomyosin contractility that is regulated by soft microenvironments (10s kPa) promotes the initiation and propagation of the long-distance ICWs. In this R35 grant, we propose a cross-disciplinary research program that systematically elucidates this novel mechanotransduction process and establishes a framework to bridge the knowledge gap. This will be accomplished by leveraging innovative approaches such as genetically encoded fluorescent calcium sensors, CRISPR imaging, high-throughput cell selection, and algorithms to pursue two interrelated research themes. The first theme is the identification of regulatory mechanisms that initiate the calcium waves by active modulations and live-cell imaging of contractility- associated proteins. The expected results will provide important molecular insights of the link between mechanotransduction and the ICWs. The second theme is the dissection of mechanisms through which calcium waves enhance cell migration and achieve biological functions by manipulating ICWs and investigating the full transcriptome profiles. The results will advance our understanding of the relationships between microenvironmental mechanics, cell signaling, gene expression, and cell functions. We envision that the fundamental principles uncovered in this project will apply broadly to various cell systems and physiological functions. The long-term objective of our research program is to establish a mechanistic foundation of mechanobiology and promote the creation of therapeutic strategies which leverage these principles. The success of this proposal will enable my group to embark in a long-term research direction to tackle a variety of critical and challenging questions regarding the interplay between mechanotransduction and cell signaling.

项目摘要 机械转导是细胞感知细胞外机械刺激并将其转化为 细胞内信号和基因表达。机械转导在不同的生物体中普遍存在， 对细胞功能和行为有重大影响，与化学和遗传信号转导平行。 微环境力学通过细胞-微环境相互作用来调节机械转导 而它的不当监管是各种病理的核心。该领域的一个主要知识空白是如何机械 来自微环境的刺激被转化为细胞信号，以及它们之间的关系 微环境力学、细胞信号、基因表达和细胞功能。我们最近发现， 多个电不可兴奋的上皮细胞系可以发起和繁殖自发的远距离 细胞在2D/3D软微环境中培养，而不是在僵硬微环境中培养时，细胞间钙波(ICW) 一个。由于钙调节广泛的基本细胞功能，我们的发现揭示了一种 微环境力学与多种细胞之间史无前例的力学转导关系 信号。我们假设，受软微环境调节的细胞肌球蛋白收缩能力 (10s Kpa)促进了长距离ICW的萌生和传播。在这笔35卢比的赠款中，我们提议 跨学科研究计划，系统地阐明这一新的机械转导过程和 建立一个弥合知识鸿沟的框架。这将通过利用创新的 基因编码的荧光钙传感器、CRISPR成像、高通量细胞等方法 选择和算法来追求两个相互关联的研究主题。第一个主题是识别 通过主动调节和收缩能力的活细胞成像来启动钙波的调节机制- 相关蛋白质。预期的结果将提供关于两者之间联系的重要分子洞察力 机械转导和ICW。第二个主题是对钙通过 WAVE通过操纵ICW和研究完整的 转录组特征。这一结果将促进我们对两者之间关系的理解 微环境力学、细胞信号、基因表达和细胞功能。我们设想， 这个项目中揭示的基本原理将广泛应用于各种细胞系统和生理 功能。我们研究计划的长期目标是建立一个机械基础 机制生物学，并促进创造利用这些原则的治疗策略。成功之路 这项建议将使我的小组能够着手进行长期的研究方向，以解决各种关键和 关于机械转导和细胞信号之间相互作用的挑战性问题。

项目成果

Biophysics in tumor growth and progression: from single mechano-sensitive molecules to mechanomedicine.

• DOI: 10.1038/s41388-023-02844-x
• 发表时间: 2023-11
• 期刊: ONCOGENE
• 影响因子: 8
• 作者: Xin, Ying;Li, Keming;Huang, Miao;Liang, Chenyu;Siemann, Dietmar;Wu, Lizi;Tan, Youhua;Tang, Xin
• 通讯作者: Tang, Xin