Data_Sheet_1_Resolvin D1 Reduces Lung Infection and Inflammation Activating Resolution in Cystic Fibrosis.docx

Non-resolving lung inflammation and Pseudomonas aeruginosa infections are the underlying cause of morbidity and mortality in cystic fibrosis (CF). The endogenous lipid mediator resolvin (Rv) D1 is a potent regulator of resolution, and its roles, actions, and therapeutic potential in CF are of interest. Here, we investigated actions and efficacy of RvD1 in preclinical models of cystic fibrosis. Cftr knockout mice with chronic P. aeruginosa lung infection were treated with RvD1 to assess differences in lung bacterial load, inflammation, and tissue damage. Cells from volunteers with CF were treated with RvD1 during ex vivo infection with P. aeruginosa, and effects on phagocytosis and inflammatory signaling were determined. In CF mice, RvD1 reduced bacterial burden, neutrophil infiltration, and histological signs of lung pathology, improving clinical scores of diseases. Mechanistically, RvD1 increased macrophage-mediated bacterial and leukocyte clearance in vivo. The clinical significance of these findings is supported by actions in primary leukocytes and epithelial cells from volunteers with CF where RvD1 enhanced P. aeruginosa phagocytosis and reduced genes and proteins associated to NF-κB activation and leukocyte infiltration. Concentration of RvD1 in sputum from patients with CF was also inversely correlated to those of cytokines and chemokines involved in CF lung pathology. These findings demonstrate efficacy of RvD1 in enhancing resolution of lung inflammation and infections and provide proof of concept for its potential as a prototypic novel pro-resolutive therapeutic approach for CF.

囊性纤维化（cystic fibrosis, CF）患者发病与死亡的根本诱因，为迁延不愈的肺部炎症与铜绿假单胞菌（Pseudomonas aeruginosa）感染。内源性脂质介质消退素（resolvin, Rv）D1是强效的炎症消退调控因子，其在囊性纤维化中的功能、作用机制与治疗潜力均受到学界广泛关注。本研究针对囊性纤维化的临床前模型，探究了RvD1的作用效果与治疗功效。本研究对携带慢性铜绿假单胞菌肺部感染的囊性纤维化跨膜传导调节因子（CFTR）敲除小鼠予以RvD1干预，以评估其肺部细菌载量、炎症状态与组织损伤的变化情况。同时，在体外模拟铜绿假单胞菌感染的实验体系中，对囊性纤维化患者来源的细胞施以RvD1处理，进而检测其对吞噬活性与炎症信号通路的影响。在囊性纤维化小鼠模型中，RvD1可降低细菌负荷、减少中性粒细胞浸润，并缓解肺部病理的组织学特征，同时改善疾病临床评分。从机制层面而言，RvD1在体内可增强巨噬细胞介导的细菌与白细胞清除能力。本研究的临床转化价值得到了囊性纤维化患者原代白细胞与上皮细胞实验结果的佐证：RvD1可增强铜绿假单胞菌的吞噬活性，并下调与核因子κB（nuclear factor kappa-light-chain-enhancer of activated B cells, NF-κB）激活及白细胞浸润相关的基因与蛋白表达。此外，囊性纤维化患者痰液中的RvD1浓度，与参与肺部病理过程的细胞因子及趋化因子水平呈显著负相关。本研究结果证实了RvD1可促进肺部炎症与感染的消退，并为其作为囊性纤维化新型原型促消退治疗手段的潜力提供了概念验证。