Mechanotransduction of shear stress: from ATP release to CFTR regulation

剪切应力的机械传导：从 ATP 释放到 CFTR 调节

• 批准号: 7447993
• 负责人: BRIAN M BUTTON
• 金额: $ 9.8万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7447993
• 关键词: Address; Agonist; Breathing; Cell membrane; Cell physiology; Cell surface; Cells; Characteristics; Chloride Channels; Cilia; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Cytoskeleton; Disease; Drowning; Elements; Epithelial; Epithelial Cells; Epithelium; Exocytosis; Fluids and Secretions; Foundations; Funding Opportunities; Goals; Hydration status; Ion Transport; Kinetics; Lead; Mechanical Stimulation; Mechanical Stress; Mechanics; Mediating; Membrane; Mentored Research Scientist Development Award; Molecular; Mucous body substance; Nature; Normal Range; Nucleotides; Pathway interactions; Physiological; Process; Property; Purinoceptor; Range; Rate; Regulation; Research; Research Design; Research Personnel; Role; Signal Transduction; Stress; Stretching; Surface; Swelling; Techniques; Testing; Transport Process; Work; airway epithelium; apical membrane; autocrine; base; career; cell type; design; experience; insight; novel therapeutics; paracrine; research study; response; shear stress; transmission process

DESCRIPTION (provided by applicant): In this application, Dr. Brian Button is proposing original research to investigate the mechanisms regulating shear stress-mediated ATP release. ATP release and autocrine/paracrine stimulation of purinergic receptors has been implicated in the regulation of a wide array of cell functions in numerous diverse cell types. A key role of purinergic signaling is the regulation of various ion transport processes, including the CFTR chloride channel. External mechanical stresses, including shear, compression, stretch, and cell swelling represent a ubiquitous mechanism to stimulate ATP release. However, the mechanisms responsible for mechanotransduction of external stresses to ATP release are unknown. Recently, the PI and collaborators discovered that the oscillatory nature of stress, such as experienced during normal breathing, is essential to stimulate ATP release. Furthermore, they found that the relationship between the magnitude of oscillatory stress and the rate of ATP release was steepest within the physiological range of normal breathing, whereas stronger forces generated weaker responses. These results lead the PI to hypothesize that cells can actively regulate the rate of ATP release during mechanical stimulation, thus protecting themselves from the potentially detrimental effect of unregulated ATP release and over-stimulation of purinoceptors. Preliminary results suggest that oscillatory stress-mediated ATP release occurs by a mechanism involving transmission of external and cilia beating-mediated forces through the cytoskeleton and exocytosis-dependent secretory pathways. The work outlined in this project is designed to systematically address several components of the mechanotransduction pathway involved in ATP release and establish its physiological role in the regulation of epithelial function. To achieve these objectives, the candidate will employ a variety of techniques grouped into three Specific Aims. Aim 1 will focus on the kinetic properties of stress-stimulated ATP release and identify the cytoskeletal elements involved in the vesicular-mediated process. Aim 2 will test the hypothesis that oscillatory shear stress of magnitude above physiological ranges reduces ATP release by altering the properties of the cell membrane. Finally, Aim 3 will test whether airway epithelia sense and respond to changes in the hydration status of the overlying mucus by internal stresses generated by cilia beating transmitted to the cytoskeleton. Together, these studies are designed to provide invaluable insights into the mechanism regulating ATP release in response to external and internal forces, which may potentially lead to the discovery of novel therapeutic approaches to modulate ATP release, important in such diseases as cystic fibrosis, where ATP release has been shown to stimulate mucus clearance. This K01 award will provide the foundation for Dr. Button to pursue his career goals of becoming an independent investigator and establishing scientific funding opportunities.

描述（由申请人提供）： 在本申请中，Brian Button 博士提出了原创性研究来调查调节剪切应力介导的 ATP 释放的机制。 ATP 释放和嘌呤能受体的自分泌/旁分泌刺激与多种不同细胞类型的多种细胞功能的调节有关。嘌呤能信号传导的关键作用是调节各种离子传输过程，包括 CFTR 氯离子通道。外部机械应力，包括剪切、压缩、拉伸和细胞肿胀，是刺激 ATP 释放的普遍机制。然而，外部压力机械转导至 ATP 释放的机制尚不清楚。最近，PI 和合作者发现压力的振荡性质（例如正常呼吸期间经历的压力）对于刺激 ATP 释放至关重要。此外，他们发现，在正常呼吸的生理范围内，振荡应力的大小与 ATP 释放速率之间的关系最为陡峭，而较强的力会产生较弱的反应。这些结果使 PI 推测细胞可以在机械刺激期间主动调节 ATP 释放速率，从而保护自身免受不受调节的 ATP 释放和嘌呤受体过度刺激的潜在有害影响。初步结果表明，振荡应激介导的 ATP 释放是通过一种机制发生的，该机制涉及通过细胞骨架和胞吐作用依赖性分泌途径传递外部和纤毛跳动介导的力。该项目概述的工作旨在系统地解决参与 ATP 释放的机械转导途径的几个组成部分，并确定其在上皮功能调节中的生理作用。为了实现这些目标，候选人将采用分为三个具体目标的各种技术。目标 1 将重点关注应激刺激 ATP 释放的动力学特性，并确定参与囊泡介导过程的细胞骨架元件。目标 2 将检验以下假设：高于生理范围的振荡剪切应力通过改变细胞膜的特性来减少 ATP 释放。最后，目标 3 将测试气道上皮是否通过纤毛跳动产生的内应力传递到细胞骨架来感知和响应上层粘液水合状态的变化。总之，这些研究旨在为响应外力和内力调节 ATP 释放的机制提供宝贵的见解，这可能会导致发现调节 ATP 释放的新治疗方法，这对于囊性纤维化等疾病很重要，其中 ATP释放已被证明可以刺激粘液清除。该 K01 奖项将为 Button 博士追求成为独立研究者和建立科学资助机会的职业目标奠定基础。