Ivacaftor for Acquired CFTR Dysfunction in Chronic Rhinosinusitis

Ivacaftor 治疗慢性鼻窦炎获得性 CFTR 功能障碍

• 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
• 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' -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

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.

无效的粘膜纤毛清除（MCC）是导致慢性炎症的常见病理生理过程。 鼻窦炎（CRS）-一种发病率很高的高度流行疾病。我们实验室的证据表明 表明，短暂暴露于革兰氏阴性菌的脂多糖（LPS）会导致 通过减少人类和其他人CFTR顶端Cl-通道的阴离子转运功能失调的MCC 哺乳动物物种。在这种情况下，CFTR抑制是由TLR 4介导的活性氧产生引起的 物种（ROS），但独立于NF κ B衍生的炎症。此外，我们的小组已经确认， 在人类鼻窦中存在获得性CFTR功能障碍，并且已经证明CFTR增效剂可以 当在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。我们 中心假设是LPS诱导的获得性CFTR缺陷，1）在很大程度上有助于 CRS的发病机制，和2）可以用Ivacaftor治疗，Ivacaftor是一种为CF开发的CFTR增效剂 疗法具体目标1将研究LPS介导的鼻腔鼻窦CFTR功能障碍的机制基础， 通过1）通过AMP依赖性激酶（抑制剂）检查CFTR的氧化剂依赖性抑制 CFTR调节结构域的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（质谱法）的影响， 和3）测量较长时间暴露于LPS对CFTR表达、成熟加工和 回收利用。目的2将在临床前兔模型中鉴定依伐卡托改善CFTR功能的效率 获得性CFTR缺陷：1）开发LPS暴露兔上颌窦CFTR的标准数据 功能障碍，2）评估依伐卡托对CFTR相关终点的影响，和3）评估依伐卡托作为 治疗铜绿假单胞菌兔上颌窦炎。Aim 3将进行一项临床研究 依伐卡托在CRS患者中的应用：1）与新型“内窥镜引导的窦电位差”测定相关 CRS疾病严重程度的验证措施和2）使用依伐卡托进行CRS的初步临床试验 难治性革兰氏阴性细菌CRS患者。目前的建议将有助于澄清机制 负责鼻窦炎的发病机制，而且还将我们的实验室发现转化为人类受试者， 提供了一种使用依伐卡托治疗鼻窦炎的临床试验。我们相信我们的应用程序将回答 关于CRS的病理机制的基本问题，并为新的 一种治疗严重和使人衰弱的慢性疾病的方法。