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

基本信息

批准号：
10226878
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
项目类别：
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/10226878
关键词：
ATAC-seq Acids Alternative Splicing Anions Apical Area Asthma Basophils Biology Bronchoscopy Brush Cell CRISPR/Cas technology Cell Culture Techniques Cells Chromatin Chronic Cultured Cells 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/antagonist 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

项目摘要

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.

项目概要/摘要 2 型炎症是通过释放 IL-33 等主细胞因子在气道上皮引发的驱动 2 型细胞因子产生、嗜酸性粒细胞增多和 2 型粘液病理学转变为炎症。当通常包含它的稳态机制失败导致持续性疾病时，就会持续存在。肺部成像（计算机断层扫描）经常显示哮喘气道中的粘液堵塞，并且这些堵塞物呈高度嗜酸性并持续多年。这些发现使我们提出气道损伤会导致。上皮细胞重新编程，导致局部区域出现 2 型炎症和粘液堵塞（“2 型气道我们有三个目标来描述哮喘 2 型生态位的生物学特征，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 将表征免疫细胞的亚型、其受体表达及其生态位特定基因我们将使用质谱流式细胞术 (CyTOF) 来计数 2 型细胞因子产生细胞及其受体。 AIM 2 将使用批量和单细胞测序来表征微环境中的上皮细胞。以及揭示这些细胞中生态位特异性表观遗传变化的方法，重点关注调节基因 2 型细胞因子（IL-33、TSLP、IL25、IL1β）和全基因组甲基化研究将包括在内。 AIM 3 将探索堵塞和非堵塞气道上皮细胞的表观遗传变化。上皮细胞和嗜酸性粒细胞之间的串扰如何导致 2 型气道中粘液栓的形成这一目标的重点将放在导致上皮细胞上调的 IL-13 调节途径上。运输氧化还原相关的卤化物（例如硫氰酸盐）并增加粘蛋白样分子的截面，例如为了实现其三个目标，项目 3 将与项目 1 和 2 密切互动，并将利用其优势。所有核心的资源，特别是人类受试者核心的资源和核心C的分析能力。该项目将通过多种方式推进对 2 型利基的了解，从而指出新的治疗策略来改变消除 2 型炎症并从根本上改善哮喘。