PDF(Pubmed)
Abstract：
BACKGROUND: Cystic Fibrosis causing mutations in the gene CFTR, reduce the activity of the CFTR channel protein, and leads to mucus aggregation, airway obstruction and poor lung function. A role for CFTR in the pathogenesis of other muco-obstructive airway diseases such as Chronic Obstructive Pulmonary Disease (COPD) has been well established. The CFTR modulatory compound, Ivacaftor (VX-770), potentiates channel activity of CFTR and certain CF-causing mutations and has been shown to ameliorate mucus obstruction and improve lung function in people harbouring these CF-causing mutations. A pilot trial of Ivacaftor supported its potential efficacy for the treatment of mucus obstruction in COPD. These findings prompted the search for CFTR potentiators that are more effective in ameliorating cigarette-smoke (CS) induced mucostasis.
METHODS: Small molecule potentiators, previously identified in CFTR binding studies, were tested for activity in augmenting CFTR channel activity using patch clamp electrophysiology in HEK-293 cells, a fluorescence-based assay of membrane potential in Calu-3 cells and in Ussing chamber studies of primary bronchial epithelial cultures. Addition of cigarette smoke extract (CSE) to the solutions bathing the apical surface of Calu-3 cells and primary bronchial airway cultures was used to model COPD. Confocal studies of the velocity of fluorescent microsphere movement on the apical surface of CSE exposed airway epithelial cultures, were used to assess the effect of potentiators on CFTR-mediated mucociliary movement.
RESULTS: We showed that SK-POT1, like VX-770, was effective in augmenting the cyclic AMP-dependent channel activity of CFTR. SK-POT-1 enhanced CFTR channel activity in airway epithelial cells previously exposed to CSE and ameliorated mucostasis on the surface of primary airway cultures.
CONCLUSIONS: Together, this evidence supports the further development of SK-POT1 as an intervention in the treatment of COPD.
METHODS: Small molecule potentiators, previously identified in CFTR binding studies, were tested for activity in augmenting CFTR channel activity using patch clamp electrophysiology in HEK-293 cells, a fluorescence-based assay of membrane potential in Calu-3 cells and in Ussing chamber studies of primary bronchial epithelial cultures. Addition of cigarette smoke extract (CSE) to the solutions bathing the apical surface of Calu-3 cells and primary bronchial airway cultures was used to model COPD. Confocal studies of the velocity of fluorescent microsphere movement on the apical surface of CSE exposed airway epithelial cultures, were used to assess the effect of potentiators on CFTR-mediated mucociliary movement.
RESULTS: We showed that SK-POT1, like VX-770, was effective in augmenting the cyclic AMP-dependent channel activity of CFTR. SK-POT-1 enhanced CFTR channel activity in airway epithelial cells previously exposed to CSE and ameliorated mucostasis on the surface of primary airway cultures.
CONCLUSIONS: Together, this evidence supports the further development of SK-POT1 as an intervention in the treatment of COPD.
摘要：
背景：囊性纤维化导致CFTR基因突变，降低CFTR通道蛋白的活性，并导致粘液聚集，气道阻塞和肺功能差。CFTR在其他粘液阻塞性气道疾病如慢性阻塞性肺疾病(COPD)的发病机理中的作用已得到充分证实。CFTR调节化合物，Ivacaftor(VX-770),增强CFTR和某些引起CF的突变的通道活性，并已被证明可以改善患有这些引起CF的突变的人的粘液阻塞并改善肺功能。Ivacaftor的初步试验支持其治疗COPD粘液阻塞的潜在疗效。这些发现促使人们寻找更有效改善香烟烟雾（CS）诱导的粘液停滞的CFTR增效剂。
方法：小分子增效剂，先前在CFTR结合研究中发现，在HEK-293细胞中使用膜片钳电生理学测试了增强CFTR通道活性的活性，在Calu-3细胞和原发性支气管上皮培养的Ussing室研究中，基于荧光的膜电位测定。向沐浴Calu-3细胞顶端表面的溶液中添加香烟烟雾提取物（CSE），并使用原发性支气管气道培养物模拟COPD。CSE暴露的气道上皮培养物的顶表面上荧光微球运动速度的共聚焦研究，用于评估增效剂对CFTR介导的粘膜纤毛运动的影响。
结果：我们表明SK-POT1与VX-770一样，可有效增强CFTR的环AMP依赖性通道活性。SK-POT-1增强了先前暴露于CSE的气道上皮细胞中的CFTR通道活性，并改善了原发性气道培养物表面的粘液淤滞。
结论：一起，这一证据支持SK-POT1作为COPD治疗的干预措施的进一步发展.
方法：小分子增效剂，先前在CFTR结合研究中发现，在HEK-293细胞中使用膜片钳电生理学测试了增强CFTR通道活性的活性，在Calu-3细胞和原发性支气管上皮培养的Ussing室研究中，基于荧光的膜电位测定。向沐浴Calu-3细胞顶端表面的溶液中添加香烟烟雾提取物（CSE），并使用原发性支气管气道培养物模拟COPD。CSE暴露的气道上皮培养物的顶表面上荧光微球运动速度的共聚焦研究，用于评估增效剂对CFTR介导的粘膜纤毛运动的影响。
结果：我们表明SK-POT1与VX-770一样，可有效增强CFTR的环AMP依赖性通道活性。SK-POT-1增强了先前暴露于CSE的气道上皮细胞中的CFTR通道活性，并改善了原发性气道培养物表面的粘液淤滞。
结论：一起，这一证据支持SK-POT1作为COPD治疗的干预措施的进一步发展.
