School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland, Dublin 2, Ireland.
National Children's Research Centre, Children's Health Ireland (CHI) at Crumlin, Dublin 12, Ireland.
Thorax. 2020 Jun;75(6):449-458. doi: 10.1136/thoraxjnl-2019-214027. Epub 2020 Apr 7.
Mutations in the cystic fibrosis transmembrane regulator () gene form the basis of cystic fibrosis (CF). There remains an important knowledge gap in CF as to how diminished CFTR activity leads to the dominant inflammatory response within CF airways.
To investigate if extracellular vesicles (EVs) contribute to inflammatory signalling in CF.
EVs released from CFBE41o-, CuFi-5, 16HBE14o- and NuLi-1 cells were characterised by nanoparticle tracking analysis (NTA). EVs isolated from bronchoalveolar lavage fluid (BALF) from 30 people with CF (PWCF) were analysed by NTA and mass spectrometry and compared with controls. Neutrophils were isolated from the blood of 8 PWCF to examine neutrophil migration in the presence of CFBE41o- EVs.
A significantly higher level of EVs were released from CFBE41o- (p<0.0001) and CuFi-5 (p=0.0209) relative to control cell lines. A significantly higher level of EVs were detected in BALF of PWCF, in three different age groups relative to controls (p=0.01, 0.001, 0.002). A significantly lower level of EVs were released from CFBE41o- (p<0.001) and CuFi-5 (p=0.0002) cell lines treated with CFTR modulators. Significant changes in the protein expression of 126 unique proteins was determined in EVs obtained from the BALF of PWCF of different age groups (p<0.001-0.05). A significant increase in chemotaxis of neutrophils derived from PWCF was observed in the presence of CFBE41o EVs (p=0.0024) compared with controls.
This study demonstrates that EVs are produced in CF airway cells, have differential protein expression at different ages and drive neutrophil recruitment in CF.
囊性纤维化跨膜转导调节因子（CFTR）基因突变是囊性纤维化（CF）的基础。CF 气道中 CFTR 活性降低如何导致占主导地位的炎症反应，这在 CF 中仍然存在重要的知识空白。
研究细胞外囊泡（EVs）是否有助于 CF 中的炎症信号传导。
通过纳米颗粒跟踪分析（NTA）对 CFBE41o-、CuFi-5、16HBE14o-和 NuLi-1 细胞释放的 EVs 进行了表征。通过 NTA 和质谱分析对来自 30 名 CF 患者（PWCF）支气管肺泡灌洗液（BALF）中分离的 EVs 进行了分析，并与对照进行了比较。从 8 名 PWCF 的血液中分离中性粒细胞，以研究 CFBE41o-EVs 存在时中性粒细胞的迁移。
CFBE41o-（p<0.0001）和 CuFi-5（p=0.0209）相对对照细胞系释放的 EVs 水平显著升高。与对照组相比，在不同年龄组的 PWCF 的 BALF 中检测到的 EVs 水平显著升高（p=0.01、0.001、0.002）。用 CFTR 调节剂处理 CFBE41o-（p<0.001）和 CuFi-5（p=0.0002）细胞系后，EVs 的释放水平显著降低。在来自不同年龄组 PWCF 的 BALF 中获得的 EVs 中，确定了 126 种独特蛋白质的表达水平发生了显著变化（p<0.001-0.05）。与对照组相比，来自 PWCF 的中性粒细胞的趋化性在 CFBE41o-EVs 的存在下显著增加（p=0.0024）。
本研究表明，EVs 在 CF 气道细胞中产生，在不同年龄时具有不同的蛋白表达，并在 CF 中驱动中性粒细胞募集。
