Ivacaftor for Acquired CFTR Dysfunction in Chronic Rhinosinusitis

Ivacaftor 治疗慢性鼻窦炎获得性 CFTR 功能障碍

基本信息

批准号：
9324344
负责人：
Bradford Alan Woodworth
金额：
$ 36.65万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9324344
关键词：
5&apos -AMP-activated protein kinase Affect Anions Apical Biological Assay Biological Models Cell Culture Techniques Cell Wall Cell surface Cells Chloride Channels Chloride Ion Chlorides Chronic Chronic Disease Clinical Clinical Protocols Clinical Research Clinical Trials Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Defect Disease Epithelial Epithelial Cells Epithelium Exposure to Foundations Functional disorder Future Generations Gram-Negative Bacteria Human Hydrogen Peroxide Impairment In Vitro Inflammation Investigational Therapies Ion Transport Laboratories Laboratory Finding Lead Lipopolysaccharides Mass Spectrum Analysis Maxillary Sinus Maxillary Sinusitis Measurement Measures Mediating Modality Modeling Morbidity - disease rate Mucociliary Clearance Mucous body substance Mus Nasal Epithelium Nose Oryctolagus cuniculus Oxidants PKA inhibitor Pathogenesis Patients Pharmaceutical Preparations Phosphorylation Pilot Projects Positioning Attribute Pre-Clinical Model Process Proteins Protocols documentation Pseudomonas aeruginosa Reactive Oxygen Species Recycling Refractory Role Scientist Secondary to Severity of illness Sinus Sinusitis Structure Superoxides Surface Surgeon TLR4 gene Testing Therapeutic Translating Viscosity chronic rhinosinusitis clinically relevant human subject improved in vivo in vivo Model indexing inflammatory lung disease kinase inhibitor novel novel therapeutic intervention novel therapeutics patch clamp patient oriented research pre-clinical respiratory standard care therapy development translational study

项目摘要

Ineffective mucociliary clearance (MCC) is a common pathophysiologic process contributing to chronic rhinosinusitis (CRS) - a highly prevalent disease with substantial morbidity. Evidence from our laboratory has demonstrated that brief exposures to lipopolysaccharide (LPS) from gram-negative bacteria lead to dysfunctional MCC by decreasing anion transport through CFTR apical Cl- channels in humans and other mammalian species. CFTR inhibition in this setting is caused by TLR4-mediated generation of reactive oxygen species (ROS), but independent of NFkB-derived inflammation. Furthermore, our group has confirmed the presence of acquired CFTR dysfunction in human sinuses, and has demonstrated that CFTR potentiators can stimulate Cl- secretion when partial CFTR dysfunction is present in multiple in vitro and preclinical models. Our central hypotheses are that LPS-induced acquired CFTR deficiency, 1) contributes substantially to the pathogenesis of CRS, and 2) can be treated with Ivacaftor, a CFTR potentiator developed for CF therapy. Specific Aim 1 will investigate the mechanistic basis of LPS-mediated CFTR dysfunction in sinonasal epithelium by 1) examining the oxidant-dependent inhibition of CFTR via AMP-dependent kinase (an inhibitor of PKA-dependent phosphorylation of the CFTR regulatory domain), 2) assessing the impact of ROS (superoxide, hydrogen peroxide) on CFTR function (patch clamp analysis) and structure (mass spectrometry), and 3) measuring the effects of longer exposures to LPS on CFTR expression, maturational processing, and recycling. Aim 2 will identify the efficiency of Ivacaftor in improving CFTR function in a pre-clinical rabbit model of acquired CFTR deficiency by 1) developing normative data for LPS-exposed rabbit maxillary sinus CFTR dysfunction, 2) assessing the effects of Ivacaftor on CFTR-related endpoints, and 3) evaluating Ivacaftor as therapy for Pseudomonas aeruginosa rabbit maxillary sinusitis. Aim 3 will conduct a clinical study using Ivacaftor in CRS patients by 1) correlating the novel “endoscopically-directed sinus potential difference” assay to validated measures of CRS disease severity and 2) performing a pilot clinical trial using Ivacaftor for CRS patients with refractory gram-negative bacterial CRS. The current proposal will help clarify mechanisms responsible for sinusitis pathogenesis, but also translates our laboratory findings to human subjects by providing a clinical trial using Ivacaftor for therapy of sinusitis. We believe our application will answer fundamental questions regarding pathomechanisms underlying CRS and establish the foundation for a new therapeutic approach to a serious and debilitating chronic disease.

无效的粘液纤毛清除（MCC）是导致慢性疾病的常见病理生理过程鼻窦炎 (CRS) - 我们实验室的证据显示，这是一种发病率很高的疾病。证明短暂暴露于革兰氏阴性细菌的脂多糖（LPS）会导致通过减少人类和其他动物通过CFTR顶端Cl-通道的阴离子转运来治疗功能失调的MCC 在这种情况下，哺乳动物物种的 CFTR 抑制是由 TLR4 介导的活性氧的产生引起的。种（ROS），但与 NFkB 衍生的炎症无关。此外，我们的小组已经证实了这一点。人类鼻窦中存在获得性 CFTR 功能障碍，并已证明 CFTR 增强剂可以当多种体外和临床前模型中存在部分 CFTR 功能障碍时，刺激 Cl- 分泌。中心假设是 LPS 诱导的获得性 CFTR 缺陷，1) 很大程度上导致了 CRS 的发病机制，以及 2) 可以使用 Ivacaftor 进行治疗，Ivacaftor 是一种针对 CF 开发的 CFTR 增强剂具体目标 1 将研究 LPS 介导的鼻窦 CFTR 功能障碍的机制基础。 1) 通过 AMP 依赖性激酶（一种抑制剂）检查 CFTR 的氧化剂依赖性抑制 CFTR 调节域的 PKA 依赖性磷酸化），2）评估 ROS 的影响（超氧化物、过氧化氢）对 CFTR 功能（膜片钳分析）和结构（质谱）的影响， 3) 测量较长时间暴露于 LPS 对 CFTR 表达、成熟加工和目标 2 将确定 Ivacaftor 在临床前兔子模型中改善 CFTR 功能的效率。通过 1) 开发暴露于 LPS 的兔上颌窦 CFTR 的规范数据来减少获得性 CFTR 缺陷功能障碍，2) 评估 Ivacaftor 对 CFTR 相关终点的影响，以及 3) 评估 Ivacaftor 作为 Aim 3将使用铜绿假单胞菌治疗兔上颌窦炎进行临床研究。 Ivacaftor 在 CRS 患者中的作用：1) 关联新型“内窥镜引导窦电位差”测定验证 CRS 疾病严重程度的衡量标准，以及 2) 使用 Ivacaftor 进行 CRS 试点临床试验目前的提议将有助于阐明难治性革兰氏阴性细菌 CRS 的机制。负责鼻窦炎的发病机制，但也将我们的实验室发现转化为人类受试者我们相信我们的应用程序会提供使用 Ivacaftor 治疗鼻窦炎的临床试验。有关 CRS 病理机制的基本问题，并为新的研究奠定基础治疗严重且使人衰弱的慢性疾病的方法。