Immune Airway-Epithelial Interactions in Steroid-Refractory Severe Asthma

重度哮喘（SA）是一种复杂的、多因素的、皮质类固醇（CS）治疗无效的疾病。近几 近年来，随着生物制剂的出现，其治疗取得了一定进展。然而，尽管这些 尽管取得了一些进展，但由于不完全的免疫缺陷，这些患者的治疗仍然存在挑战。 了解免疫和上皮细胞的功能障碍，这是这种疾病的基础。这进一步 复杂的不良生物标志物可用于区分疾病的靶向治疗，并放大， 他们的巨大成本，特别是在规定不准确的情况下。2001年项目1取得的进展 使用质谱细胞术/CyTOF的循环显示SA患者聚类为2组，显示不同的免疫反应。 分布图沿着先天免疫和适应性免疫的线划分。项目2在以下方面取得的进展 SA中的支气管上皮细胞（BEC）表明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，以驱动先天性的 内在表型在第二组中，BEC细胞死亡途径与CD 8 T细胞免疫交叉， 驱动免疫相互作用表型的过程。综上所述，这些发现促使我们 假设有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，是SA的关键决定因素。气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诱导细胞死亡导致两种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 使用人类样品和小鼠疾病模型：a）表征CITE-seq和TCR-seq气道 通过BAL收集免疫细胞，并研究dupilumab对免疫表型的影响，B）研究 表达FceRI的先天性免疫细胞和FceRI+细胞中的IL-7 Ra信号传导在促进SA中的重要性 使用新的基于蛋白酶的小鼠模型研究表型，和c）研究调节的潜在机制， TRM细胞表型和使用SA的T细胞驱动的小鼠模型对疾病的影响。项目2将使用新鲜的 和培养的健康和哮喘BEC，以：a）确定 先天性内在上皮表型，B）评价GSDMB在CD 8 T细胞发育中的作用 免疫相互作用表型及其功能意义，和c）整合免疫-炎症 表型与离体和体外上皮分子表型。项目之间的协同互动 将由核心A（行政核心）提供支持，以协调该计划的活动 各级项目，核心B，人体生物取样和免疫细胞计数核心和核心C， RNA-Seq和生物信息学核心。总之，这些项目将在免疫领域建立新的概念- 上皮细胞相互作用是哮喘发病机制的基础，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.