Departments of Biochemistry and Physiology, The Hospital for Sick Children, Programs in Cell Biology and Neuroscience and Mental Health, and University of Toronto, Toronto, ON, Canada M5G 1X8.
Proc Natl Acad Sci U S A. 2011 Feb 22;108(8):3216-21. doi: 10.1073/pnas.1010334108. Epub 2011 Feb 7.
Cystic fibrosis is caused by impaired ion transport due to mutated cystic fibrosis transmembrane conductance regulator, accompanied by elevated activity of the amiloride-sensitive epithelial Na(+) channel (ENaC). Here we show that knockout of the ubiquitin ligase Nedd4L (Nedd4-2) specifically in lung epithelia (surfactant protein C-expressing type II and Clara cells) causes cystic fibrosis-like lung disease, with airway mucus obstruction, goblet cell hyperplasia, massive inflammation, fibrosis, and death by three weeks of age. These effects of Nedd4L loss are likely caused by enhanced ENaC function, as reflected by increased ENaC protein levels, increased lung dryness at birth, amiloride-sensitive dehydration of lung explants, and elevated ENaC currents in primary alveolar type II cells analyzed by patch clamp recordings. Moreover, the lung defects were rescued with administration of amiloride into the lungs of young knockout pups via nasal instillation. Our results therefore suggest that the ubiquitin ligase Nedd4L can suppress the onset of cystic fibrosis symptoms by inhibiting ENaC in lung epithelia.
囊性纤维化是由囊性纤维化跨膜电导调节因子突变导致的离子转运受损引起的,伴随着阿米洛利敏感的上皮钠离子通道(ENaC)活性升高。在这里,我们表明,在肺上皮细胞(表面活性蛋白 C 表达的 II 型和克拉拉细胞)中特异性敲除泛素连接酶 Nedd4L(Nedd4-2)会导致囊性纤维化样肺部疾病,表现为气道黏液阻塞、杯状细胞增生、大量炎症、纤维化,并在 3 周龄时死亡。Nedd4L 缺失的这些影响可能是由于 ENaC 功能增强引起的,这反映在 ENaC 蛋白水平升高、出生时肺干燥度增加、肺外植体中阿米洛利敏感的脱水以及通过膜片钳记录分析的原代肺泡 II 型细胞中 ENaC 电流升高。此外,通过鼻腔滴注将阿米洛利递送至年轻的敲除幼崽肺部,可以挽救肺部缺陷。因此,我们的研究结果表明,泛素连接酶 Nedd4L 通过抑制肺上皮细胞中的 ENaC 可以抑制囊性纤维化症状的发生。