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

项目摘要/摘要 2型炎症是通过释放主要细胞因子如IL-33在呼吸道上皮细胞启动的 驱动2型细胞因子的产生、嗜酸性粒细胞增多和粘液病理。2型炎症变成 当正常情况下包含它的动态平衡机制失败时，持续存在，导致持续疾病。我们发现 肺部成像(计算机断层扫描)经常显示粘液堵塞在哮喘的呼吸道中，并且堵塞 是高度嗜酸性的，并持续多年。这些发现导致我们提出，呼吸道损伤会导致 对上皮重新编程以引起2型炎症和粘液堵塞的局部区域(“2型呼吸道”) 利基“)。我们有三个目标来描述哮喘中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新编程以及IL-13驱动的粘液塞子形成机制。 目标1将描述免疫细胞的亚型、它们的受体表达和它们的生态位特异性基因 表情。我们将使用质量细胞术(CyTOF)来计数产生2型细胞因子的细胞及其受体 表达曲目。AIM 2将使用批量和单细胞测序来表征壁龛中的上皮细胞 也有方法发现这些细胞中特定于小生境的表观遗传学变化，重点放在调节 2型细胞因子(IL-33、TSLP、IL-25、IL-1β)。将包括atac-seq和全基因组甲基化研究。 研究堵塞和未堵塞的呼吸道上皮细胞的表观遗传学变化。《目标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型呼吸道粘液堵塞的形成 利基市场。这一目标的重点将放在IL-13调节的导致上皮细胞上调的途径上 运输与氧化还原相关的卤化物，如硫氰酸盐，并增加粘蛋白类分子的截面，如 FCγBP.为了实现其三个目标，项目3将与项目1和项目2密切互动，并将利用 在所有核心中，特别是人类学科核心的资源和核心C的分析能力。 该项目将以可能指向新的治疗策略的方式促进对2型利基的了解 消除II型炎症，从根本上改善哮喘。

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.