Effects of shear stress on regulation of epithelial permeability

剪切应力对上皮通透性调节的影响

基本信息

批准号：
8100234
负责人：
Venkataramana K Sidhaye
金额：
$ 13.55万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-03 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8100234
关键词：
Actins Air Apical Area Asthma Breathing Calcium Cations Cells Coughing Critical Care Cytoskeleton DNA Sequence Rearrangement Data Development Disease Endothelial Cells Environment Environmental air flow Epidermal Growth Factor Epithelial Epithelial Cells Event Exercise Foundations Frequencies Goals Height Hepatocyte Human Infection Inflammation Ion Channel Kidney L-Type Calcium Channels Lung Mechanics Medicine Membrane Metabolic acidosis MicroRNAs Modeling Mus Nifedipine Nitric Oxide Organ Pathologic Pathway interactions Permeability Physiological Physiology Play Production Property Proteins Regulation Relative (related person)Reporting Research Personnel Respiration Respiratory physiology Rest Role Signal Transduction Source Stimulus Structure of respiratory epithelium Surface TRPV channel Techniques Tidal Volume Tight Junctions Trachea Tubular formation Vanilloid Vascular Endothelial Cell Water abstracting airway epithelium airway surface liquid aquaporin 5 career claudin-1 protein cytokine experience genetic manipulation human NOS3 protein in vitro Model inhibitor/antagonist interest novel particle programs receptor release of sequestered calcium ion into cytoplasm response shear stress skills tool voltage gated channel water channel

项目摘要

DESCRIPTION (provided by applicant): The goal of this K08 application is to facilitate development of essential skills that will allow the PI to become a successful academician and achieve independent scientific investigator status. The respiratory epithelium serves as a barrier, a regulator of the content of airway surface liquid, and a source of cytokines and other products that regulate airway physiology. In addition to cyclic extensions and contraction, airway epithelial cells are exposed to lumenal shear stress, defined as the frictional force per unit surface area, generated by airflow. The shear stress sensed by airway epithelia is continuously changing under both physiologic and pathologic conditions. We have exciting novel data suggesting that shear stress in airway epithelial cells leads to changes in airway epithelial permeability. Therefore, dynamic modulation of the airway epithelial barrier could be an important mechanism by which epithelial cells transduce lumenal information into subepithelial responses. Our preliminary studies indicate that shear stress induces TRPV4 activation with subsequent L-type voltage gated channel activation with subsequent increases in intracellular calcium. The increase in intracellular calcium in airway epithelial cells leads to concerted changes in epithelial paracellular permeability by modulating actin rearrangement and NO production, as well as in transmembrane permeability by modulating AQP5 abundance. We propose to study the role of shear stress in altering both paracellular and transmembrane permeability as well as dissect the underlying mechanisms regulating these effects. In SA#1we will investigate the effects of shear stress on airway epithelial permeability using primary cultured human airway epithelial cells (NHBE) and.ex vivo isolated mouse trachea. In SA#2 we will define the role for and the mechanisms (TRPV4, L-type channel) of shear-induced calcium flux in modulating epithelial permeability using pharmacologic inhibition and genetic manipulation. In SA#3 we will study the role od shear-induced increases in calcium on NO production and its effect on permeability. Through these complimentary techniques and coursework, the PI will develop new skills and generate novel data regarding the effects of mechanical shear stress on airway epithelial cell regulation and function, a little-examined area likely to be of immense importance to normal lung function, as well as conditions of altered shear including exercise and asthma. (End of Abstract)

描述（由申请人提供）： K08 申请的目标是促进基本技能的发展，使 PI 成为一名成功的院士并获得独立科学研究员的地位。呼吸道上皮充当屏障、气道表面液体含量的调节器以及细胞因子和其他调节气道生理学的产物的来源。除了周期性的伸展和收缩之外，气道上皮细胞还承受腔内剪切应力，定义为气流产生的每单位表面积的摩擦力。气道上皮感受的剪切应力在生理和病理条件下不断变化。我们有令人兴奋的新数据表明气道上皮细胞中的剪切应力导致气道上皮通透性的变化。因此，气道上皮屏障的动态调节可能是上皮细胞将管腔信息转换为上皮下反应的重要机制。我们的初步研究表明，剪切应力诱导 TRPV4 激活，随后 L 型电压门控通道激活，随后细胞内钙含量增加。气道上皮细胞中细胞内钙的增加通过调节肌动蛋白重排和NO产生导致上皮细胞旁通透性的协调变化，以及通过调节AQP5丰度导致跨膜通透性的协调变化。我们建议研究剪切应力在改变细胞旁通透性和跨膜通透性方面的作用，并剖析调节这些效应的潜在机制。在 SA#1 中，我们将使用原代培养的人气道上皮细胞 (NHBE) 和离体分离的小鼠气管来研究剪切应力对气道上皮通透性的影响。在 SA#2 中，我们将定义剪切诱导的钙流在使用药理学抑制和遗传操作调节上皮通透性中的作用和机制（TRPV4，L 型通道）。在 SA#3 中，我们将研究剪切诱导的钙增加对 NO 产生的作用及其对渗透性的影响。通过这些免费的技术和课程，PI 将开发新技能并生成关于机械剪切应力对气道上皮细胞调节和功能的影响的新数据，这是一个很少被研究的领域，可能对正常肺功能也非常重要作为改变剪切力的条件，包括运动和哮喘。（摘要完）