Amphotericin B Restoration of Anion Secretion in Cystic Fibrosis Airways

两性霉素 B 恢复囊性纤维化气道阴离子分泌

基本信息

批准号：
10248529
负责人：
MICHAEL J. WELSH
金额：
$ 47.53万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10248529
关键词：
Aerosols Amphotericin B Animals Anions Anti-Bacterial Agents Apical Bacterial Infections Bicarbonates Binding Cells Clinical Clinical Research Collaborations Cyclic AMP Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Defect Development Disease Distal Environment Epithelial Epithelial Cells Exhibits Family suidae Future Genes Genotype Gland Goals Height Heterogeneity Host Defense Human Illinois Impairment Inflammation Iowa Knowledge Label Learning Lung Mediating Molecular Mucociliary Clearance Mucous body substance Mutation Mycoses Nebulizer Pathogenesis Play Property Pulmonary Cystic Fibrosis Regulation Research Research Personnel Respiratory Failure Role Safety Staphylococcus aureus Sterols Surface Testing Viscosity Work airway epithelium airway remodeling airway surface liquid antimicrobial apical membrane cell type cystic fibrosis airway cystic fibrosis airway epithelia improved in vivo insight loss of function mutation mutant null mutation programs pulmonary function repaired respiratory restoration success treatment strategy

项目摘要

PROJECT 2 PROJECT SUMMARY Cystic fibrosis (CF) is caused by loss-of-function mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) apical membrane anion channel. Loss of CFTR- mediated HCO3- and Cl- secretion by airway epithelia impairs respiratory host defenses, causing bacterial infection, inflammation, mucus accumulation, and respiratory failure. One strategy for restoring anion channel function to the apical membrane is to increase the function of mutant CFTR channels. That has proven successful for people with specific CFTR mutations. However, ~10% of people with CF have mutations that do not respond to CFTR modulators or they are not able to take modulators. Another strategy is to provide an alternative channel. Toward this end, we studied amphotericin B (AmB); earlier work showed that AmB forms anion channels. We discovered that apical AmB increased HCO3- and Cl- secretion in cultured CF airway epithelia, including those with CFTR-null mutations. AmB increased ASL pH, height, and antibacterial activity. AmB is clinically approved to treat fungal infections, and off-label lung aerosolization has an impressive safety record. Thus, AmB could potentially provide a new mutation-agnostic therapy for CF lung disease. However, many questions remain unanswered and we lack important in vivo data. Therefore, our overarching goal is to understand the mechanisms of AmB-induced anion secretion in airway epithelia and to test the hypothesis that AmB can restore CF host defenses in vivo. To achieve this goal, we will answer questions in three aims. Aim 1. What molecular and cellular mechanisms elicit AmB-mediated anion secretion? Without cAMP-dependent regulation and with half the anion selectivity of CFTR, AmB is an imperfect substitute for CFTR. We will test key hypotheses about how AmB functions in airway epithelia. Aim 2. Which epithelial cells does AmB target and how does AmB alter them? These studies will reveal AmB function in large and small airway epithelia and diverse cell-types, test AmB in airway epithelia remodeled by inflammation, and test for compensatory changes. Aim 3. Does AmB reverse CF host defense defects in vivo? Using CF pigs, we will test if nebulized AmB increases ASL pH, increases ASL antimicrobial activity and height, increases mucociliary transport, and enhances S. aureus eradication from the lungs. The results of these studies will have direct implications for developing a new genotype agnostic approach to treating CF, and will inform development of other therapies in Projects 1 and 3. Our proposal’s success is enhanced by an ongoing collaboration between the labs of Welsh (Iowa) and Burke (Illinois), by utilization of outstanding cores, and by an environment of cooperation between the Program’s projects and investigators. With this background, our track record, and our commitment, we believe this research can change the lives of people with CF.

项目2 项目概要囊性纤维化 (CF) 是由编码囊性纤维化的基因发生功能丧失突变引起的跨膜电导调节器 (CFTR) 顶膜阴离子通道丧失 CFTR-。气道上皮介导的 HCO3- 和 Cl- 分泌损害呼吸道宿主防御，导致细菌感染感染、炎症、粘液积聚和呼吸衰竭是恢复负离子的一种策略。顶膜通道的功能是增加突变CFTR通道的功能。已证明对具有特定 CFTR 突变的人有效。然而，约 10% 的 CF 患者患有这种疾病。对 CFTR 调节剂没有反应或不能接受另一种调节剂的突变。为此，我们早期研究了两性霉素B（AmB）；工作表明 AmB 形成阴离子通道我们发现顶端 AmB 增加了 HCO3- 和培养的 CF 气道上皮细胞（包括具有 CFTR 缺失 AmB 突变的细胞）中的 Cl- 分泌。 AmB 增加 ASL pH、高度和抗菌活性，经临床批准可用于治疗真菌感染、因此，AmB 可能会提供一个令人印象深刻的安全记录。然而，许多问题仍未得到解答和解决。我们缺乏重要的体内数据，因此，我们的首要目标是了解其机制。 AmB 诱导气道上皮细胞分泌阴离子，并检验 AmB 可以恢复的假设 CF 主机体内防御为了实现这一目标，我们将回答三个目标 1. 目标是什么。分子和细胞机制引发 AmB 介导的阴离子分泌，而不依赖于 cAMP？ AmB 具有 CFTR 一半的阴离子选择性，是 CFTR 的完美替代品。测试有关 AmB 如何在空气上皮细胞中发挥作用的关键假设目标 2。哪些上皮细胞发挥作用。 AmB 的目标以及 AmB 如何改变它们？这些研究将揭示 AmB 的功能。气道上皮和多种细胞类型，测试炎症重塑的气道上皮中的 AmB，并测试目标 3. AmB 是否可以利用 CF 猪逆转 CF 宿主防御缺陷？我们将测试雾化 AmB 是否会增加 ASL pH 值、增加 ASL 抗菌活性和高度，增加粘液纤毛运输，并增强金黄色葡萄球菌从肺部的根除效果。这些研究将对开发一种新的基因型不可知论治疗方法产生直接影响 CF，并将为项目 1 和 3 中其他疗法的开发提供信息。我们提案的成功是威尔士（爱荷华州）和伯克（伊利诺伊州）实验室之间的持续合作得到了加强，利用优秀的核心，并通过该计划项目之间的合作环境有了这样的背景、我们的记录和我们的承诺，我们相信这项研究。可以改变 CF 患者的生活。