Immune Airway-Epithelial Interactions in Steroid-Refractory Severe Asthma

重症哮喘(SA)是一种复杂的、多因素的疾病，对糖皮质激素(CS)治疗无效。在最近 多年来，随着生物制剂的出现，在其治疗方面取得了一些进展。然而，尽管如此， 尽管取得了进展，但由于不完全的原因，在这些患者的治疗方面仍然存在挑战 了解免疫和上皮细胞的功能障碍，这是这种疾病的基础。这是更远的 由于可用于靶向治疗的可用于区分疾病的糟糕的生物标志物而变得复杂，并被 它们的巨大成本，特别是在处方不准确的情况下。当前项目1取得的进展 用质量细胞仪/细胞周期图显示SA患者聚集成两组，表现出明显的免疫 按照先天免疫和获得性免疫的路线划分的特征。项目2的研究进展 SA中的支气管上皮细胞(BECs)提示存在两种根本不同的分子 表型。一组中的BEC主要对吸入的环境刺激做出反应，以驱动天生的 内在表型。在第二组中，BEC细胞死亡途径与CD8 T细胞免疫相交 驱动免疫互动表型的过程。综上所述，这些发现促使我们 假设两种不同的免疫机制，一种由先天免疫细胞调节，另一种由T细胞调节 细胞，是SA的关键决定因素。呼吸道上皮细胞之间的相互作用 遗传/表观遗传风险和免疫细胞以及细胞死亡的诱导导致两个SA 表型。这一假设将在以下两个高度互动的项目中得到解决：项目1将 使用人类样本和小鼠疾病模型：a)表征CITE-SEQ和TCR-SEQ的呼吸道 BAL收集的免疫细胞，并研究DUPILUMA对免疫表型的影响，b)研究 FceRI阳性细胞表达FceRI和IL-7ra信号在促进SA发生中的作用 使用一种新的基于蛋白酶的小鼠模型的表型，以及c)研究潜在的调控机制 使用T细胞驱动的SA小鼠模型研究TRM细胞表型及其对疾病的影响。项目2将使用Fresh 并培养健康和哮喘的BEC以：a)确定其机制和功能意义 先天固有的上皮表型，b)评估GSDMB在CD8 T细胞发育中的作用 免疫相互作用表型及其功能意义；c)免疫-炎症一体化 表型与上皮分子表型的体外和体外研究。项目之间的协同互动 将由核心A(行政核心)提供支助，以协调方案的活动 各级项目，核心B、人类生物采样和免疫细胞计数核心和核心C、 RNA-Seq和生物信息学核心。综上所述，这些项目将在免疫领域建立新的概念 哮喘发病机制中的上皮相互作用，并确定新的治疗靶点。

Severe asthma (SA) is complex, multifactorial and refractory to treatment by corticosteroids (CS). In recent years, some progress has been made in its treatment with the advent of biologics. However, despite these advancements, there still remain challenges in the treatment of these patients because of an incomplete understanding of the dysfunction of immune and epithelial cells, which underlies this disease. This is further complicated by the poor biomarkers available to differentiate disease for targeted therapy, and amplified by their enormous costs, particularly when prescribed imprecisely. Progress made in Project 1 in the current cycle using mass cytometry/CyTOF, show clustering of SA patients into 2 groups displaying distinct immune profiles divided along the lines of innate and adaptive immunity. Progress made in Project 2 in studies of bronchial epithelial cells (BECs) in SA suggest the presence of two fundamentally different molecular phenotypes. The BECs in one group primarily respond to inhaled environmental stimuli to drive an innate intrinsic phenotype. In the second group, BEC cell death pathways intersect with CD8 T-cell immune processes to drive an immune interactive phenotype. Taken together, these findings prompted us to hypothesize that two distinct immune mechanisms, one regulated by innate immune cells, and the second by T cells, are critical determinants of SA. Interactions between airway epithelial cells in the context of genetic/epigenetic risks and immune cells together with induction of cellular death contribute to two SA phenotypes. This hypothesis will be addressed in the following two highly interactive projects: Project 1 will use human samples and mouse models of disease to: a) characterize the CITE-seq and TCR-seq airway immune cells collected by BAL and investigate the impact of dupilumab on immune phenotype, b) study the importance of FceRI-expressing innate immune cells and IL-7Ra signaling in FceRI+ cells in promoting a SA phenotype using a novel protease-based mouse model, and c) study mechanisms underlying regulation of TRM cell phenotype and impact on disease using a T cell-driven mouse model of SA. Project 2 will use fresh and cultured healthy and asthmatic BECs to: a) determine the mechanisms for and functional implications of an innate intrinsic epithelial phenotype, b) evaluate the role of GSDMB in the development of a CD8 T-cell immune interactive phenotype and its functional implications, and c) Integrate immune-inflammatory phenotypes with epithelial molecular phenotypes ex vivo and in vitro. Synergistic interactions among projects will be afforded by support from: Core A, the Administrative Core, to coordinate the activities of the Program Project at all levels, Core B, the Human Biological Sampling and Immunocytometry Core and Core C, the RNA-Seq and Bioinformatics Core. Taken together, these projects will establish new concepts in immune- epithelial interactions underlying asthma pathogenesis and identify novel targets for therapy.