Initiators, biomarkers and mechanisms of epithelial dysfunction and immune pathogenesis in chronic rhinosinusitis and aspirin exacerbated respiratory disease (AERD)

项目摘要 慢性鼻窦炎（CRS）导致生活质量的严重损失和超过100亿美元的费用给美国 医疗保健系统每年。治疗选择主要限于皮质类固醇、抗生素和手术。 CRS通常分为两种主要形式，一种没有鼻息肉形成（CRSsNP），另一种有鼻息肉形成（CRSsNP）。 鼻息肉形成（CRSwNP）。CRSwNP的一种特别严重的形式是阿司匹林急性呼吸道感染。 疾病（AERD）。AERD患者患有鼻息肉病和哮喘，通常是严重的形式。的 这一提议要检验的基本假设是，免疫屏障中的缺陷组合沿着 炎症和结构细胞的活化驱动CRS和AERD的发病机制。次中心 该建议的目的包括提高我们对CRSwNP和AERD发病机理的认识， 开发一种新的基于微粒（MP）的工具，用于诊断AERD并评估其重要性 激素和细胞因子在CRS发病机制中的上皮活化中的作用。我们将使用一个大的， 对因CRS接受手术的患者进行的稳健研究以及基于尖端实验室和生物信息学的研究 研究CRS致病机制的方法。拟议的工作分为三个具体目标。在 第一个目标，我们将测试假设上皮间质转化（EMT）是一个标志性特征， CRSwNP反映了潜在的内分泌紊乱，并与CRSwNP的严重程度和结局相关。 我们将CRSwNP、AERD和对照组的上皮刮片中EMT的程度与疾病终点联系起来 为了验证我们的假设，即激素变化、制瘤素M和上皮调节素在CRS中起重要作用- 相关的EMT和CRS发病机制。在第二个目标中，我们将测试炎症性 白细胞、结构细胞和肥大细胞在CRSwNP和AERD中被激活，并导致症状， 疾病活动。我们将使用一种创新的MP检测方法，结合流式细胞术和RNASeq（Drop-Seq）， 评估对照组中嗜碱性粒细胞、嗜酸性粒细胞、肥大细胞、血小板、内皮细胞和上皮细胞的活化 和CRSwNP或AERD患者。我们将把细胞激活与使用CT评估的疾病严重程度联系起来， 内窥镜检查、治疗史、问卷调查和组织病理学。我们将炎症终点与EMT联系起来 重要的临床终点。最后，我们将检验嗜酸性粒细胞活化程度越高， 嗜碱性粒细胞、肥大细胞和血小板解释了AERD中疾病的更严重性。在目标三中，我们将使用 MP检测用于在CRSwNP患者的大队列中识别未诊断的AERD患者， 共病哮喘我们将通过阿司匹林激发试验确认或排除AERD的诊断，以验证MP试验。 这些研究将进一步了解CRS和AERD的发病机制，并为我们提供更多的信息。 更接近于新的诊断和治疗干预。

PROJECT SUMMARY Chronic rhinosinusitis (CRS) causes profound loss of quality of life and over $10B of expense to the US healthcare system annually. Treatment options are primarily limited to corticosteroids, antibiotics and surgery. CRS is conventionally divided into two major forms, one without nasal polyp formation (CRSsNP), and one with nasal polyp formation (CRSwNP). A particularly severe form of CRSwNP is Aspirin Exacerbated Respiratory Disease (AERD). Patients with AERD suffer from both nasal polyposis and asthma, usually in severe forms. The fundamental hypothesis to be tested by this proposal is that a combination of defects in the immune barrier along with activation of inflammatory and structural cells drives pathogenesis in CRS and AERD. Secondary central goals of this proposal include advancement of our knowledge of pathogenesis of CRSwNP and AERD, development of a new microparticle (MP) based tool for diagnosis of AERD and assessment of the importance of hormones and cytokines in epithelial activation in CRS pathogenesis. We will use a combination of large, robust studies of patients undergoing surgery for CRS and cutting edge laboratory-based and bioinformatics approaches to study CRS pathogenic mechanisms. The proposed work is divided into three specific aims. In the first aim, we will test the hypothesis that epithelial-mesenchymal transition (EMT) is a hallmark feature of CRSwNP reflecting underlying endocrine disturbances and associates with severity and outcomes in CRSwNP. We will relate the extent of EMT in epithelial scrapings from CRSwNP, AERD and controls to disease endpoints to test our hypotheses that hormonal changes, Oncostatin M and Epiregulin play important roles in CRS- associated EMT and CRS pathogenesis. In the second aim, we will test the hypothesis that inflammatory leukocytes, structural cells and mast cells are activated in CRSwNP and AERD and contribute to symptoms and disease activity. We will use an innovative MP assay combined with flow cytometry and RNASeq (Drop-Seq) to assess activation of basophils, eosinophils, mast cells, platelets, endothelial cells and epithelial cells in controls and patients with CRSwNP or AERD. We will relate cell activation to disease severity assessed using CT, endoscopy, treatment history, questionnaires and tissue pathology. We will link inflammatory endpoints to EMT and important clinical endpoints. Finally, we will test the hypothesis that greater activation of eosinophils, basophils, mast cells and platelets explains the greater severity of disease in AERD. In aim three, we will use the MP assay to identify patients with undiagnosed AERD among a large cohort of patients with CRSwNP and comorbid asthma. We will confirm or eliminate diagnosis of AERD with aspirin challenge to validate the MP test. These studies will advance knowledge of the mechanisms of pathogenesis of CRS and AERD and bring us closer to new diagnostics and therapeutic interventions.