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.

该提案的主要目标是（i）确定由 铜质细胞化生（GCM）和粘液分泌的铜绿假单胞菌（PA） 纤维化（CF）气道，（ii）表征了诱导机制，（iii）确定其对CF的重要性 发病。 PA是急性和慢性肺部感染的常见原因，包括CF和非CF 支气管扩张，呼吸机相关肺炎（VAP）和慢性阻塞性肺疾病（COPD）。 PA感染与这些患者的发病率和死亡率显着增加有关。之一 这些患病的肺部共有的主要病理特征在于其失调，粘液分泌过度 清除中的故障，导致气道堵塞，可减少气体交换并恶化肺功能。 众所周知，PA可以分泌各种二级代谢物，包括VOC。近年来，微生物VOC 在患者的呼吸中发现已被审查为疾病诊断的潜在生物标志物，包括 CF，VAP和COPD。不幸的是，VOC对肺发病机理的生物学作用，包括GCM和 粘液过度分泌，知之甚少。在初步研究中，我们证明了几个主要的 以前在患有CF的人的呼吸中鉴定出的PA VOC种（例如2-氨基乙烯酮，2-AA） 在人气道上皮细胞和小鼠中诱导GCM和粘液过度分泌。在此应用程序中，我们将 测试假说是VOC是患病气道中GCM和粘液分泌的主要因素。 AIM 1将单独和能够诱导GCM的鸡尾酒中的主要PA VOC物种来表征 正常人原发支气管上皮细胞（NHBEC）与CF疾病初级的粘液分泌 在常氧和低氧下，在空气界面（ALI）中培养的支气管上皮细胞（CF-DHBEC） 条件分别模仿正常和CF气道。顶级VOC将进行单独检查或 在鸡尾酒中，它们能够诱导野生型与B-ENAC过表达的GCM和粘液过度分泌的能力 （SCNN1B-TG）CF小鼠。最后，均野生型PA菌株PA14及其等源性DPQSE突变体在2-AA中缺乏 将比较它们在野生型与B-ENAC小鼠中诱导GCM和粘液过度分泌的能力。目的 2将检查PA VOC的GCM和粘液生物合成诱导的机制。首先，我们将确定 VOC是否激活Pro-GCM途径，包括AHR和EGFR-AKT/ERK1/2，以抑制表达 FOXA2，气道粘液稳态的关键调节器。然后，我们将检查VOC是否也引起GCM和 通过调节巨噬细胞极化，中性粒细胞流入和Th1/Th2/Th17/ILC2S免疫来调节粘液生物合成 反应是患病气道中GCM和粘液过度分泌的已知驱动因素。完成 提案将揭示CF和其他患病肺中VOC与过度粘液之间的机械联系， 并且，潜在的针对细菌性肺炎的新型治疗方法。