INVESTIGATION OF SALT CONCENTRATION IN THE AIRWAY SURFACE LIQUID USING X-RAY FLU

使用 X 射线流感病毒研究气道表面液体中的盐浓度

• 批准号: 7598301
• 负责人: GERARD C WONG
• 金额: $ 0.02万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598301
• 关键词: Affect; Cell Line; Computer Retrieval of Information on Scientific Projects Database; Custom; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Defect; Disease; Epithelial; Epithelial Cells; Event; Excision; Fluids and Secretions; Fluorescence; Funding; Grant; Human; Hydration status; Institution; Investigation; Ion Channel; Ion Transport; Ions; Lead; Liquid substance; Live Birth; Lung; Measurement; Measures; Molecular; Monitor; Mucous body substance; Mutation; Numbers; Pathogenesis; Relative (related person); Research; Research Personnel; Resources; Respiratory Failure; Sampling; Sodium Chloride; Source; Synchrotrons; Thick; Time; United States National Institutes of Health; Viscosity; airway surface liquid; apical membrane; base; chelation; culture plates; flu

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There is no cure for Cystic Fibrosis (CF), a lethal disease that affects 1 of every 3,000 live births in the U.S. In CF, mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), a Cl ion channel found in the apical membranes of secretory epithelial cells, disrupt epithelial ion transport and result in the accumulation of viscous infected mucus in pulmonary airways which lead to respiratory failure. The degree of mucus hydration, which determines the mucus viscosity, is abnormal in CF, and a number of competing explanations have been proposed. The only way to discriminate between these different mechanisms is a measurement of the salt level in the airway surface liquid (ASL), a 10 micron thick layer of fluid covering airway epithelial cells, to see if it has been increased or decreased relative to normal levels. Although this is the most important open question in CF, it is extremely difficult to answer reliably. A number of approaches have been attempted, such as conductivity measurements, direct ASL removal, and fluorescent chelation compounds as ion concentration indicators, but the results vary over an order of magnitude. The principal difficulty lies in the fact that any contact measurement will induce a dramatic increase in fluid secretion and change the measured salt concentration. We propose a synchrotron-based non-contact measurement of the salt level in the ASL. We are the only group that has successfully cultured human airway epithelial cell lines both with and without the CF defect. These samples can be mounted on custom-made culture plates. Using GXF, grazing x-ray fluorescence, it will be possible to simultaneously monitor the concentrations of all the relevant ion species involved in the CF pathogenesis, and establish the sequence of molecular events which is responsible for the disease for the first time, as well as detailed pharmacological studies.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 囊性纤维化(CF)是一种致命的疾病，在美国每3,000名活产儿中就有1例受到影响。囊性纤维化(CF)是一种位于分泌上皮细胞顶膜上的氯离子通道，囊性纤维化跨膜传导调节因子(CFTR)的突变扰乱了上皮离子的运输，导致粘性感染粘液在肺内积聚，从而导致呼吸衰竭。决定粘液粘度的粘液水合程度在CF中是异常的，并提出了许多相互竞争的解释。区分这些不同机制的唯一方法是测量呼吸道表面液体(ASL)中的盐分水平，ASL是覆盖在呼吸道上皮细胞上的一层10微米厚的液体，看看它相对于正常水平是增加了还是减少了。虽然这是CF中最重要的开放问题，但要可靠地回答它是极其困难的。已经尝试了许多方法，例如电导率测量、直接去除ASL和作为离子浓度指示剂的荧光络合化合物，但结果在一个数量级上有所不同。主要的困难在于，任何接触式测量都会导致液体分泌量急剧增加，并改变测量的盐浓度。我们提出了一种基于同步加速器的非接触测量ASL中盐度的方法。我们是唯一一个成功培养出有和没有CF缺陷的人呼吸道上皮细胞系的小组。这些样品可以安装在定制的培养皿上。利用GXF，掠射X射线荧光，将有可能同时监测参与CF发病机制的所有相关离子物种的浓度，并首次建立导致这种疾病的分子事件序列，以及详细的药理学研究。