Anti-inflammatory therapy to augment CFTR rescue in CF patients

抗炎治疗可增强 CF 患者的 CFTR 救援

• 批准号: 9753341
• 负责人: Matthias A Salathe
• 金额: $ 65.5万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-18 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753341
• 关键词: Address; Adhesions; Air; Angiotensin II Receptor; Angiotensin Receptor; Animal Model; Anti-inflammatory; Antiinflammatory Effect; Apical; Birth; Calcium; Cells; Chloride Channels; Clinical; Cultured Cells; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Defect; Development; Disease; Drug Kinetics; Effectiveness; Epithelial Cells; Event; Excision; FDA approved; Failure; Family suidae; Functional disorder; Genetic Diseases; Heart Diseases; Human; Hypertension; Impairment; In Vitro; Inflammation; Inhalation; Intervention; Ion Transport; Laboratories; Leukocyte Elastase; Life; Link; Liquid substance; Literature; Longevity; Losartan; Lung; Lung diseases; Measurement; Measures; Mediating; Modeling; Mucociliary Clearance; Mucous body substance; Mutation; Nose; Oral; Outcome; PPAR gamma; Patient Care; Patients; Pharmaceutical Preparations; Potassium Channel; Recovery; Regulation; Respiratory physiology; Sheep; Signal Transduction; Testing; Transforming Growth Factors; absorption; airway inflammation; airway surface liquid; cystic fibrosis airway; cystic fibrosis patients; cytokine; driving force; epithelial Na+ channel; experimental study; improved; improved functioning; in vivo; knock-down; large-conductance calcium-activated potassium channels; lung preservation; mutant; novel; novel strategies; pre-clinical; prevent; small molecule; success; voltage

Project Summary The life of patients with Cystic fibrosis (CF) continues to be shortened by progressive lung disease. Recently, the development of specific small molecules that rescue mutation specific CFTR function have made remarkable impacts on patient care, but they are not available for many patients. In addition, ongoing airway inflammation may reduce their effectiveness, at least in some patients. In this context, TGF-b1, a cytokine associated with worse pulmonary outcome in CF, decreases airway surface liquid (ASL) volume in CF cells in vitro by impairing the function of calcium activated chloride channels (CaCCs) and the apical potassium channel BK, both crucial for maintaining ASL volume in CF. TGF-b1 also reduced mucociliary clearance in sheep. We therefore propose that airway inflammation in general and TGF-b1 in particular worsens mucociliary function in CF patients. If this hypothesis is correct, novel anti-inflammatory medications could improve CF outcomes in patients currently not eligible for CFTR recovery therapy on which we focus here, but might also improve the function of small molecules. Clinically used Angiotensin II Receptor Blockers (ARBs) inhibit TGF-b signaling, independent of their angiotensin receptor blocking ability. Losartan, one widely used ARB, exerts these latter effects through one of its major metabolites, EXP3179. Thus, ARBs could present a “fast track approach” for improving mucociliary dysfunction in CF. The preclinical, in vitro proof of concept aim 1 will test the hypothesis that losartan reverses the negative effects of TGF-b1 on CaCC and BK function (BK mainly through preventing decreases in LRRC26, the g regulatory subunit of BK required for its function in non- excitatory cells). Mechanisms of losartan's actions on CaCC and BK function will be explored using Ussing chambers as well as ASL volume and mucus transport measurements in fully differentiated airway epithelial cells from CF patients in the presence or absence of TGF-b1, losartan, and its metabolites EXP3174 and 3179. The preclinical, in vivo animal model aim 2 will test the hypothesis that inhaled or oral losartan improves CF- related mucociliary dysfunction in sheep induced by inhaled CFTRinh172 and human neutrophil elastase (HNE) or TGF-b1, which is downstream of HNE. We will measure tracheal mucus velocity, determine key pharmacokinetic parameters of losartan, and explore mechanisms that induce mucociliary dysfunction. Finally, the clinical in vivo aim 3 will test the hypothesis that oral losartan improves mucociliary dysfunction and decreases airway and systemic inflammation in CF patients who are not on small molecule treatment. The proposed experiments will test the ability of available anti-inflammatory medications, namely losartan and its metabolites, to improve parameters of mucociliary function in CF in vitro and in vivo and delineate their mode of action, thereby potentially identifying a “low-hanging-fruit” intervention for CF airway inflammation.

项目概要 囊性纤维化 (CF) 患者的生命继续因进行性肺部疾病而缩短。最近， 拯救突变特定CFTR功能的特定小分子的开发使得 对患者护理产生显着影响，但许多患者无法获得这些服务。此外，持续气道 炎症可能会降低其有效性，至少在某些患者中是如此。在这种情况下，TGF-b1（一种细胞因子） 与 CF 中较差的肺部结果相关，CF 细胞中气道表面液体 (ASL) 体积的减少 通过损害钙激活氯离子通道 (CaCC) 和顶端钾的功能来体外 通道 BK，两者对于维持 CF 中的 ASL 音量都至关重要。 TGF-b1 还降低了粘液纤毛清除率 羊。因此，我们认为气道炎症，特别是 TGF-b1 会恶化粘液纤毛 CF 患者的功能。如果这个假设正确，新型抗炎药物可以改善 CF 目前不符合 CFTR 恢复治疗条件的患者的结果，我们在此重点关注，但也可能 改善小分子的功能。临床使用的血管紧张素 II 受体阻滞剂 (ARB) 抑制 TGF-b 信号传导，独立于其血管紧张素受体阻断能力。氯沙坦是一种广泛使用的 ARB，可发挥 后者通过其主要代谢物之一 EXP3179 产生作用。因此，ARB 可以提供“快速通道” 方法“改善 CF 中的粘膜纤毛功能障碍。临床前、体外概念验证目标 1 将进行测试 氯沙坦逆转 TGF-b1 对 CaCC 和 BK 功能的负面影响的假设（BK 主要 通过防止 LRRC26 的减少，LRRC26 是 BK 的 g 调节亚基，在非 兴奋细胞）。将使用 Ussing 探索氯沙坦对 CaCC 和 BK 功能的作用机制 完全分化的气道上皮中的气室以及 ASL 体积和粘液转运测量 来自 CF 患者的细胞，存在或不存在 TGF-b1、氯沙坦及其代谢物 EXP3174 和 3179。 临床前体内动物模型目标 2 将检验吸入或口服氯沙坦改善 CF-的假设 吸入CFTRinh172和人中性粒细胞弹性蛋白酶（HNE）引起的绵羊相关粘液纤毛功能障碍 或 TGF-b1，它是 HNE 的下游。我们将测量气管粘液速度，确定关键 氯沙坦的药代动力学参数，并探讨诱导粘膜纤毛功能障碍的机制。最后， 临床体内目标 3 将检验口服氯沙坦改善粘膜纤毛功能障碍的假设 减少未接受小分子治疗的 CF 患者的气道和全身炎症。这 拟议的实验将测试现有抗炎药物的能力，即氯沙坦及其 代谢物，以改善 CF 体内和体外粘液纤毛功能的参数并描绘其模式 的行动，从而有可能确定 CF 气道炎症的“容易实现的成果”干预措施。