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: 7634525
• 负责人: BRIAN M BUTTON
• 金额: $ 10.01万
• 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/7634525
• 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; 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 therapeutic intervention; paracrine; research study; response; shear stress; transmission process; 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; 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 therapeutic intervention; 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）： 在此应用程序中，Brian Button博士正在提出原始研究，以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广泛细胞功能有关。嘌呤能信号传导的关键作用是调节包括CFTR氯化物通道在内的各种离子传输过程。外部机械应力（包括剪切，压缩，拉伸和细胞肿胀）代表了刺激ATP释放的无处不在的机制。但是，尚不清楚导致外部应力机械转移到ATP释放的机制。最近，PI和合作者发现，压力的振荡性质（例如正常呼吸过程中经历）对于刺激ATP释放至关重要。此外，他们发现振荡应力的大小与ATP释放速率之间的关系最陡峭，在正常呼吸的生理范围内，而强力较强的力会产生较弱的反应。这些结果导致PI假设细胞可以在机械刺激过程中主动调节ATP释放速率，从而保护自己免受不受监管的ATP释放和Purinoceptors过度刺激的潜在有害作用。初步结果表明，振荡应力介导的ATP释放是通过涉及通过细胞骨架和胞吐依赖性分泌途径传播外部和纤毛介导的力的机制而发生的。该项目中概述的工作旨在系统地解决与ATP释放有关的机械转导途径的几个组成部分，并在调节上皮功能中确立其生理作用。为了实现这些目标，候选人将采用各种技术分为三个特定目标。 AIM 1将重点放在应力刺激的ATP释放的动力学特性上，并确定囊泡介导过程中涉及的细胞骨架元素。 AIM 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降低ATP的释放。最后，AIM 3将测试气道上皮是否有意义，并通过纤毛击败传播给细胞骨架而产生的内部应力对上覆盖粘液的水合状态的变化做出反应。总之，这些研究旨在提供对调节ATP释放的机制的宝贵见解，该机制响应外部和内部力，这可能导致发现了调节ATP释放的新型治疗方法，这在囊性纤维化等疾病中很重要，在此疾病中，ATP释放已显示出刺激粘液清除的ATP释放。该K01奖将为Button博士提供基础，以追求他成为独立调查员并建立科学资金机会的职业目标。