Mechanistic triggers of excessive mucus during lung infections by bacterial volatile organic compounds

细菌挥发性有机化合物肺部感染期间粘液过多的机制触发因素

• 批准号: 10666271
• 负责人: Gee W Lau
• 金额: $ 22.23万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-25 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10666271
• 关键词: Acute; Acute Disease; Air; Airway Disease; Anabolism; Antibiotics; Aryl Hydrocarbon Receptor; Bacterial Infections; Bacterial Pneumonia; Biological; Blocking Antibodies; Bronchiectasis; CRISPR/Cas technology; Cell Differentiation process; Cell Proliferation; Cells; Chloride Channels; Chronic; Chronic Disease; Chronic Obstructive Pulmonary Disease; Chronic lung disease; Clinical Pathology; Complement; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Cystic Fibrosis sputum; Data; Deterioration; Disease; ESKAPE pathogens; Epidermal Growth Factor Receptor; Epithelial Cells; Escalator; Exposure to; Failure; Fluorescence Microscopy; Gases; Genetic Transcription; Goals; Goblet Cells; Homeostasis; Human; Hydrogen Cyanide; Hypoxia; Immune; Immune response; Individual; Infection; Inflammatory Response; Knock-out; Link; Liposomes; Liquid substance; Lung; Lung diseases; Lung infections; Lymphoid Cell; MAPK3 gene; MUC5AC gene; MUC5B gene; Macrophage; Mediating; Metaplasia; Molecular; Morbidity - disease rate; Mucins; Mucous body substance; Mus; Osmosis; Pathogen detection; Pathogenesis; Pathogenicity; Pathologic; Pathway interactions; Patients; Persons; Physiological; Population; Primary Ciliary Dyskinesias; Process; Production; Proto-Oncogene Proteins c-akt; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Pulmonary Cystic Fibrosis; Regulatory Pathway; Role; Signal Pathway; T-Lymphocyte; Testing; Thiocyanates; Western Blotting; airway epithelium; airway surface liquid; bronchial epithelium; combat; cystic fibrosis airway; cystic fibrosis mouse; cystic fibrosis patients; diagnostic biomarker; disease diagnosis; improved; microbial; microbial colonization; mortality; mucus clearance; mucus hypersecretion; mutant; neutrophil; novel strategies; novel therapeutic intervention; overexpression; pathogen; pathogenic bacteria; patient prognosis; potential biomarker; prevent; pulmonary function; respiratory colonization; secondary metabolite; ventilator-associated pneumonia; volatile organic compound

The main objectives of this proposal are to (i) identify volatile organic compounds (VOCs) produced by Pseudomonas aeruginosa (PA) that cause goblet cell metaplasia (GCM) and mucus hypersecretion in Cystic Fibrosis (CF) airways, (ii) characterize the mechanism of induction, and (iii) determine its importance to CF pathogenesis. PA is a frequent cause of acute and chronical lung infections, including CF and non-CF bronchiectasis, ventilator-associated pneumonia (VAP), and chronic obstructive pulmonary disease (COPD). Infections by PA are associated with a significant increase in morbidity and mortality in these patients. One of the major pathological features shared by these diseased lungs is in their dysregulated, hypersecretion of mucus and failure in clearance, resulting in clogged airways that reduces gas exchanges and deteriorates lung function. PA is known to secrete a variety of secondary metabolites, including the VOCs. In recent years, microbial VOCs detected in the breath of patients have been scrutinized as potential biomarkers for disease diagnosis, including CF, VAP and COPD. Unfortunately, the biological effects of VOCs on lung pathogenesis, including GCM and mucus hypersecretion, are poorly understood. In preliminary studies, we demonstrate that several dominant species of PA VOCs previously identified in the breath of people with CF (e.g., 2-aminoacetophenone, 2-AA) induce GCM and mucus hypersecretion in human airway epithelial cells, and in mice. In this application, we will test the hypothesis that VOCs are major contributors of GCM and mucus hypersecretion in the diseased airways. Aim 1 will characterize major PA VOC species individually and in a cocktail that are capable of inducing GCM and mucus secretion in normal human primary bronchial epithelial cells (NHBECs) vs. CF diseased primary bronchial epithelial cells (CF-DHBECs) cultured in the air-liquid interface (ALI), under normoxic and hypoxic conditions, mimicking normal and CF airways, respectively. Top VOCs will be further examined individually or in cocktail for their ability to induce GCM and mucus hypersecretion in wild-type vs. b-ENAC overexpressing (Scnn1b-Tg) CF mice. Finally, both wild-type PA strain PA14 and its isogenic DpqsE mutant deficient in 2-AA will be compared for their ability to induce GCM and mucus hypersecretion in wild-type vs. b-ENAC mice. Aim 2 will examine mechanisms of GCM and mucus biosynthesis induction by PA VOCs. First, we will determine whether VOCs activate pro-GCM pathways, including AhR and EGFR-AKT/ERK1/2, to inhibit the expression of FOXA2, a key regulator of airway mucus homeostasis. Then, we will examine if VOCs also induce GCM and mucus biosynthesis by modulating macrophage polarization, neutrophil influx and Th1/Th2/Th17/ILC2s immune responses, which are known drivers of GCM and mucus hypersecretion in diseased airways. Completion of the proposal will reveal the mechanistic link between VOCs and excessive mucus in CF and other diseased lungs, and, potentially novel therapeutic approaches against bacterial pneumonias.

这项建议的主要目标是(I)识别由以下物质产生的挥发性有机化合物(VOC) 引起囊性杯状细胞化生和粘液高分泌的铜绿假单胞菌 纤维化(CF)的呼吸道，(Ii)表征诱导的机制，以及(Iii)确定其对CF的重要性 发病机制。肺炎支原体是急性和慢性肺部感染的常见原因，包括慢性支气管炎和非支气管炎。 支气管扩张、呼吸机相关肺炎(VAP)和慢性阻塞性肺疾病(COPD)。 在这些患者中，PA感染与发病率和死亡率的显著增加有关。其中之一 这些病变肺的主要病理特征是粘液分泌失调。 以及清除失败，导致呼吸道堵塞，减少气体交换，恶化肺功能。 已知PA可分泌多种次生代谢物，包括VOCs。近年来，微生物VOCs 在患者呼吸中检测到的已被仔细检查为潜在的疾病诊断生物标志物，包括 Cf、VAP和COPD。不幸的是，VOCs在肺部发病机制中的生物学作用，包括GCM和 粘液高分泌，人们对此知之甚少。在初步研究中，我们证明了几个主要的 先前在CF患者呼气中发现的PA VOCs种类(例如，2-氨基苯乙酮、2-AA) 诱导人呼吸道上皮细胞和小鼠的GCM和粘液高分泌。在此应用程序中，我们将 测试VOCs是GCM和病变呼吸道粘液高分泌的主要贡献者的假设。 目标1将单独和在鸡尾酒中描述能够诱发GCM的主要PA VOC物种的特征 正常人原代支气管上皮细胞(NHBECs)与病变原代CFECs的粘液分泌 常氧和低氧条件下气液界面(ALI)培养的支气管上皮细胞 条件，分别模拟正常和CF航空。顶级挥发性有机化合物将进一步单独或 在鸡尾酒中诱导GCM和粘液高分泌野生型与b-enac过表达 (Scnn1b-Tg)CF小鼠。PA14野生型菌株及其同基因DpqsE突变体均缺乏2-AA 将比较它们在野生型和b-ENAC小鼠中诱导GCM和粘液高分泌的能力。目标 2将研究PA VOCs诱导GCM和粘液生物合成的机制。首先，我们将确定 VOCs是否激活促GCM通路，包括AhR和EGFR-AKT/ERK1/2，以抑制 FOXA2，呼吸道粘液动态平衡的关键调节因子。然后，我们将研究VOCs是否也会诱发GCM和 调节巨噬细胞极化、中性粒细胞内流和Th1/Th2/Th17/ILC2s免疫的粘液生物合成 反应，这是已知的GCM和疾病呼吸道粘液高分泌的驱动因素。已完成的 该提案将揭示挥发性有机化合物与CF和其他病变肺中过量粘液之间的机制联系， 还有，可能是治疗细菌性肺炎的新方法。