Raw data of experiments showed in Figures.

Cystic fibrosis (CF) is the most common genetic diseases in the Caucasian population. CFTR defects, the most common being F508del, lead to abnormal mucus accumulation. Respiratory failure caused by the resulting chronic infections is the leading cause of death in people with cystic fibrosis (pwCF). Pseudomonas aeruginosa is a major pathogen in CF and is responsible for a deterioration of respiratory function in pwCF. The increase of antibiotic-resistant P. aeruginosa strains encourages the search for alternative therapeutics for treating P. aeruginosa infection. In vitro studies have shown an interest in (R)-roscovitine (roscovitine) in the fight against bacterial infection in pwCF. Here we show a nuanced effect of roscovitine on ASL bactericidal activity and CF bronchial epithelium protection against P. aeruginosa. Using a 3D model of fully differentiated and functional F508del-CFTR human bronchial epithelium, we evidenced (i) an enhancement of the bactericidal activity of the airway surface liquid for 25 μM roscovitine but (ii) no limitation of the dynamic of the epithelium destruction upon roscovitine treatment whatever the concentrations. Our findings shed light on reasons for the lack of beneficial effects to prevent P. aeruginosa infection in pwCF treated with roscovitine in the ROSCO-CF clinical trial. We anticipate that our findings would have significant therapeutic implications in seeking to optimize roscovitine analogs.囊性纤维化（Cystic fibrosis, CF）是高加索人群中最为常见的遗传性疾病。囊性纤维化跨膜传导调节因子（CFTR）缺陷——其中最常见的突变类型为F508del——可引发黏液异常积聚。由此诱发的慢性感染所导致的呼吸衰竭，是囊性纤维化患者（pwCF）的首要致死诱因。铜绿假单胞菌（Pseudomonas aeruginosa）是CF患者体内的主要病原菌，可导致囊性纤维化患者的呼吸功能进行性恶化。抗生素耐药铜绿假单胞菌菌株的增多，推动了针对该菌感染的替代疗法的研发。 体外研究已证实，(R)-罗司他丁（roscovitine）在对抗囊性纤维化患者的细菌感染方面具有潜在应用价值。本研究揭示了罗司他丁对气道表面液体（airway surface liquid, ASL）杀菌活性以及支气管上皮抵御铜绿假单胞菌的保护作用所存在的差异化调控效应。我们采用完全分化且功能完备的F508del-CFTR突变型人支气管上皮三维模型，证实了两项结论：其一，25 μM浓度的罗司他丁可增强气道表面液体的杀菌活性；其二，无论使用何种浓度的罗司他丁，均无法抑制上皮损伤的动态进程。本研究结果阐明了ROSCO-CF临床试验中，接受罗司他丁治疗的囊性纤维化患者未能获得预防铜绿假单胞菌感染的有益效果的潜在原因。我们预期本研究结果可为罗司他丁类似物的优化研发提供重要的治疗学参考。