Epithelial cell reprogramming and mucus gel pathology in self-sustaining type 2 airway niches in asthma

哮喘自我维持型 2 型气道微环境中的上皮细胞重编程和粘液凝胶病理学

• 批准号: 10472542
• 负责人: John V Fahy
• 金额: $ 56.26万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10472542
• 关键词: ATAC-seq; Acids; Alternative Splicing; Anions; Apical; Area; Asthma; Basophils; Biology; Bronchoscopy; Brush Cell; CRISPR/Cas technology; Cells; Chromatin; Chronic; Cytokine Gene; Cytokine Receptors; Cytometry; Development; Disease; Disulfides; Elasticity; Eosinophilia; Epigenetic Process; Epithelial; Epithelial Cells; Functional disorder; Gel; Gene Expression; Gene Silencing; Genes; Genetic Polymorphism; Genetic Transcription; Goals; Heterogeneity; Human; Human Resources; Hydrogen Peroxide; IL18 gene; IL1R1 gene; Immune; Immunophenotyping; Individual; Inflammation; Interleukin-1 beta; Interleukin-13; Knowledge; Lead; Legal patent; Ligands; Lung; Lung CAT Scan; Mediator of activation protein; Methods; Methylation; Mucins; Mucous body substance; Nature; Outcome Measure; Oxidation-Reduction; Oxides; Pathologic; Pathology; Pathway interactions; Patients; Plug-in; Production; Property; Proteins; Role; Sampling; Signal Transduction; Sputum; TSLP gene; Thiocyanates; Tissues; Transcript; Up-Regulation; X-Ray Computed Tomography; airway epithelium; asthmatic; asthmatic airway; biophysical properties; cell type; chromatin remodeling; curative treatments; cytokine; eosinophil; eosinophil peroxidase; experimental study; human subject; inhibitor; injured airway; knock-down; lung imaging; mast cell; novel; oxidation; prevent; receptor; receptor expression; single cell sequencing; single-cell RNA sequencing; small molecule; treatment strategy; virtual; whole genome

Project Summary / Abstract Type 2 inflammation is initiated at the airway epithelium through the release of master cytokines such as IL-33 that drive type 2 cytokine production, eosinophilia, and mucus pathology. Tyope 2 inflammation becomes persistent when homeostatic mechanisms that normally contain it fail causing persistent disease. We find that lung imaging (computed tomography) frequently reveals mucus plugging in asthmatic airways and that the plugs are highly eosinophilic and persist for many years. These findings lead us to propose that airway injury leads to reprogramming of the epithelium to cause focal areas of type 2 inflammation and mucus plugging (“type 2 airway niches”). We have three Aims to characterize the biology of type 2 niches in asthma with an emphasis on reprogramming of immune cells and epithelial cells and on IL-13 driven mechanisms of mucus plug formation. AIM 1 will characterize the subtypes of immune cell, their receptor expression, and their niche specific gene expression. We will use mass cytometry (CyTOF) to enumerate type 2 cytokine producing cells and their receptor expression repertoire. AIM 2 will character epithelial cells in the niche using bulk and single cells sequencing and also methods to uncover niche-specific epigenetic changes in these cells with a focus on genes that regulate type 2 cytokines (IL-33, TSLP, IL25, IL1β). ATAC-seq and whole genome methylation studies will be included to characterize epigenetic changes in epithelial cells from plugged and non-plugged airways. AIM 3 will explore how cross-talk between epithelial cells and eosinophils results in mucus plug formation in the type 2 airway niche. Emphasis in this aim will be placed on IL-13 regulated pathways that caused epithelial cells to upregulate transport of redox-relevant halides such as thiocyanate and to increase section of mucin-like molecules such as FcγBP. To achieve its three aims, Project 3 will interact closely with projects 1 and 2, and it will take advantage of all cores, especially the resources of the human subjects core and the analytic capabilities of Core C. Our project will advance knowledge of the type 2 niche in ways that could point to novel treatment strategies to switch off type 2 inflammation and fundamentally modify asthma.

项目总结/摘要 2型炎症通过释放主要细胞因子如IL-33在气道上皮起始 其驱动2型细胞因子产生、嗜酸性粒细胞增多和粘液病理学。类型2炎症变为 当通常含有它的内稳态机制失败时持续存在，导致持续性疾病。我们发现 肺成像（计算机断层扫描）经常显示哮喘气道中的粘液堵塞， 是高度嗜酸性的并持续多年。这些发现使我们提出气道损伤导致 上皮细胞的重编程引起2型炎症和粘液堵塞的焦点区域（“2型气道 壁龛”）。我们有三个目标来描述哮喘2型小生境的生物学特征，重点是 免疫细胞和上皮细胞的重编程以及IL-13驱动的粘液栓形成机制。 AIM 1将描述免疫细胞的亚型，它们的受体表达，以及它们的小生境特异性基因 表情我们将使用质量细胞仪（CyTOF）来计数2型细胞因子产生细胞及其受体 表达谱AIM 2将使用批量和单细胞测序来表征小生境中的上皮细胞 以及揭示这些细胞中小生境特异性表观遗传变化的方法，重点是调节 2型细胞因子（IL-33、TSLP、IL 25、IL 1 β）。将纳入ATAC-seq和全基因组甲基化研究 以表征来自堵塞和非堵塞气道的上皮细胞中的表观遗传变化。AIM 3将探索 上皮细胞和嗜酸性粒细胞之间的相互作用如何导致2型气道中粘液栓的形成 利基在这个目标的重点将放在IL-13调节途径，导致上皮细胞上调 转运氧化还原相关的卤化物，如硫氰酸盐，并增加粘蛋白样分子， FcγBP。为了实现这三个目标，项目3将与项目1和项目2密切互动，并利用 尤其是人类主体核心的资源和C核心的分析能力。我们 该项目将以可能指向新的治疗策略的方式推进对2型利基的了解， 从根本上改变哮喘。