PPG -Airway Physiology and Pathophysiology in a Porcine CF Model

PPG - 猪 CF 模型中的气道生理学和病理生理学

• 批准号: 8322346
• 负责人: MICHAEL J. WELSH
• 金额: $ 227.24万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8322346
• 关键词: Affect; Animal Model; Animals; Apical; Area; Biochemical; Clinical; Collaborations; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Defect; Disease; Distal; Educational process of instructing; Electrolytes; Epithelial; Family suidae; Foundations; Functional disorder; Gland; Human; In Vitro; Infection; Inflammation; Ion Transport; Ions; Learning; Liquid substance; Longevity; Lung; Lung diseases; Measurement; Methods; Modeling; Morbidity - disease rate; Mucociliary Clearance; Mus; Pathogenesis; Phenotype; Physiological; Physiology; Process; Regulator Genes; Research; Solid; Source; System; Testing; Time; airway epithelium; airway surface liquid; clinical phenotype; cystic fibrosis airway; cystic fibrosis mouse; in vivo; insight; mortality; novel; postnatal; programs; voltage

Despite advances in CF research, we still do not understand the pathogenesis of airways disease. A major mpediment to progress is lack of a CF animal model other than the mouse. Although CF mice have been produced, they do not develop the airway disease typically found in humans. Therefore, we developed a pig with a targeted disruption of the CFTR gene. We chose the pig because its lungs share many anatomical, histological, biochemical, and physiologic features with human lungs. In this project, we take the unique opportunity to learn how loss of CFTR alters airway epithelial function in this new CF model. Several hypotheses about the pathogenesis of CF airway disease center on defective airway epithelial electrolyte transport and abnormal airway surface liquid volume and composition. These topics are the main focus of our application. Through collaborations with the other projects in the Program, we will discern how altered epithelial function relates to inflammation and infection, clinical hallmarks of the disease. We concentrate on early postnatal and young pigs because there is a critical lack of information about the human CF lung during this time, and yet this is precisely when loss of CFTR initiates disease. Specific Aim 1. Does loss of CFTR alter the function of porcine airway epithelia? We will learn how lack of CFTR changes ion transport in vivo using measurements of transepithelial voltage, in vitro using cultures of differentiated pig airway epithelia, in freshly excised airway epithelia, and in the distal airways of the lung. We will also learn whether apical Na+ channel and alternative CI" channel activities are increased, and how their function relates to the clinical phenotype. Specific Aim 2. Does loss of porcine CFTR change the airway surface liquid (ASL)? Much controversy surrounds hypotheses about how lack of CFTR affects ASL. Does it reduce ASL volume? Does it change ASL ion concentrations? Is ASL pH altered? Do changes occur both in vitro and in vivo? We will answer these questions using several independent methods. By studying both young pigs and animals after they develop inflammation and/or infection, we will discover how these processes change this critical liquid. Specific Aim 3. Does loss of CFTR disrupt mucociliary transport? Lack of CFTR might cause defective mucociliary transport (MCT) thereby initiating airway disease. Alternatively, other factors might initiate the disease process, and then secondary defects in MCT might worsen airways disease. Current data do not allow us to discriminate between these or other hypotheses. Here, we test the hypothesis that loss of CFTR disrupts normal MCT in vitro and in vivo. These studies will provide new insights into both CF pathogenesis and pathophysiology and thereby accelerate the discovery of novel therapies for this lethal disease.

尽管CF研究取得了进展，但我们仍然不了解气道疾病的发病机制。一个专业 进展的障碍是缺乏除小鼠以外的 CF 动物模型。虽然 CF 小鼠已 产生后，它们不会患上人类常见的呼吸道疾病。因此，我们开发了一种猪 有针对性地破坏 CFTR 基因。我们选择猪是因为它的肺有许多相同的解剖结构， 人肺的组织学、生化和生理特征。在这个项目中，我们采用了独特的 在这个新的 CF 模型中，有机会了解 CFTR 的丧失如何改变气道上皮功能。一些 CF气道疾病发病机制的假说集中在气道上皮电解质缺陷 运输和异常气道表面液体的量和成分。这些主题是主要关注点 我们的应用程序。通过与该计划中其他项目的合作，我们将了解如何改变 上皮功能与炎症和感染以及该疾病的临床特征有关。我们专注于 产后早期和幼猪，因为在生猪生长过程中严重缺乏有关人类 CF 肺的信息 这一次，然而这正是 CFTR 缺失引发疾病的时候。 具体目标 1. CFTR 的缺失是否会改变猪气道上皮的功能？我们将学习如何 CFTR 的缺乏改变了体内离子传输，使用跨上皮电压测量，体外使用 分化的猪气道上皮、新鲜切除的气道上皮以及猪远端气道的培养物 肺。我们还将了解顶端Na+通道和替代CI”通道活动是否增加， 以及它们的功能如何与临床表型相关。 具体目标 2. 猪 CFTR 的丧失是否会改变气道表面液体 (ASL)？争议颇多 围绕缺乏 CFTR 如何影响 ASL 的假设。它会减少 ASL 的音量吗？有变化吗 ASL 离子浓度？ ASL pH 值会改变吗？体外和体内都会发生变化吗？我们会回答 这些问题使用几种独立的方法。通过研究幼猪和成年后的动物 发生炎症和/或感染，我们将发现这些过程如何改变这种关键液体。 具体目标 3.CFTR 的丧失是否会扰乱粘膜纤毛运输？缺乏 CFTR 可能会导致缺陷 粘液纤毛运输（MCT）从而引发气道疾病。或者，其他因素可能会引发 疾病过程，然后 MCT 的继发性缺陷可能会加重气道疾病。目前的数据不 让我们能够区分这些或其他假设。在这里，我们测试 CFTR 损失的假设 破坏体外和体内正常的 MCT。 这些研究将为 CF 发病机制和病理生理学提供新的见解，从而 加速发现这种致命疾病的新疗法。