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

• 批准号: 10024667
• 负责人: MICHAEL J. WELSH
• 金额: $ 48.9万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 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/10024667
• 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; Epithelium; 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; Respiratory physiology; 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; repaired; respiratory; restoration; success; treatment strategy

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 项目摘要 囊性纤维化（CF）是由编码囊性纤维化基因的功能丧失突变引起的 跨膜电导调节剂（CFTR）顶膜阴离子通道。 CFTR-丢失 气道上皮的介导的HCO3和CL-分泌损害呼吸道防御，导致细菌 感染，感染，粘液积累和呼吸衰竭。恢复阴离子的一种策略 顶端膜的通道功能是增加突变CFTR通道的功能。那就是 对于具有特定CFTR突变的人来说是成功的。但是，约有10％的CF患者 对CFTR调节器不响应的突变或无法采用调节器。其他 策略是提供替代渠道。为此，我们研究了两性霉素B（AMB）；较早 工作表明AMB形成阴离子渠道。我们发现顶端AMB增加了HCO3-和 培养的CF气道上皮中的Cl分泌，包括具有CFTR无突变的cl分泌。 amb ASL pH，高度和抗菌活性增加。 AMB在临床上批准治疗真菌感染， 标签外肺部气化具有令人印象深刻的安全记录。那，AMB可能会提供 新的针对CF肺部病的突变 - 敏捷疗法。但是，许多问题仍未解决，并且 我们缺乏体内数据。因此，我们的总体目标是了解 AMB诱导的气道上皮分泌的阴离子分泌，并检验AMB可以恢复的假设 CF主机防御在体内。为了实现这一目标，我们将以三个目标回答问题。目标1。什么 分子和细胞机制引起了Amb介导的阴离子秘密？没有营地依赖 调节和一半的CFTR阴离子选择性，AMB是CFTR的不完善的替代品。我们将 测试关键假设关于AMB在气道上皮中的功能。 AIM 2。上皮细胞进行 AMB目标以及AMB如何改变它们？这些研究将揭示大小的AMB功能 气道上皮和各种细胞类型，通过炎症重塑气道上皮的测试AMB和测试 用于补偿性变化。 AIM 3。AMB会在体内反向CF主机防御缺陷吗？使用CF猪 我们将测试雾化的AMB是否增加ASL pH，增加ASL抗菌活性和高度， 增加粘毛的运输，并增强肺部根除金黄色葡萄球菌。结果 这些研究将对开发一种新的基因型不可知性方法具有直接影响 CF，并将告知项目1和3中其他疗法的开发。我们的提议的成功是 威尔士（爱荷华州）和伯克（伊利诺伊州）的实验室之间的持续合作加强了 利用杰出的核心，并通过该计划项目之间的合作环境 和调查人员。有了这个背景，我们的往绩和我们的承诺，我们相信这项研究 可以改变CF人的生活。