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

项目总结 慢性鼻-鼻窦炎(CRS)给美国造成严重的生活质量损失和超过100亿美元的费用 每年的医疗保健系统。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，一种是无鼻息肉形成(CRSsNP)，另一种是 鼻息肉形成(CRSwNP)。CRSwNP的一种特别严重的形式是阿司匹林加重呼吸 疾病(AERD)。AERD患者同时患有鼻息肉和哮喘，通常情况下病情严重。这个 这一提议要检验的基本假设是，免疫屏障中的缺陷 炎症细胞和结构细胞的激活推动了CRS和AERD的发病。二级中心 这项建议的目标包括促进我们对CRSwNP和AERD发病机制的了解， 一种新的基于微粒(MP)的AERD诊断工具的开发及其重要性的评估 激素和细胞因子在CRS发病机制中上皮激活中的作用。我们将结合使用大型、 对接受CRS手术患者的稳健研究以及基于实验室和生物信息学的前沿研究 CRS致病机制的研究方法。拟议的工作分为三个具体目标。在 首先，我们将检验上皮-间充质转化(EMT)是 CRSwNP反映潜在的内分泌紊乱，并与CRSwNP的严重程度和预后相关。 我们将把CRSwNP、AERD和对照的上皮刮片中EMT的程度与疾病终点联系起来 为了验证荷尔蒙变化的假说，肿瘤抑素M和表观调节素在CRS中发挥重要作用。 EMT与CRS发病机制相关。在第二个目标中，我们将检验炎症性假说 白细胞、结构细胞和肥大细胞在CRSwNP和AERD中被激活，并与症状和 疾病活动。我们将使用一种创新的MP检测结合流式细胞术和RNAseq(Drop-Seq)来 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激活情况 以及CRSwNP或AERD患者。我们将细胞激活与使用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。我们将把炎症性终点与EMT联系起来 和重要的临床终点。最后，我们将检验这样一种假设，即嗜酸性粒细胞的更大激活， 嗜碱性粒细胞、肥大细胞和血小板解释了AERD疾病的更严重程度。在目标三中，我们将使用 MP试验在大量CRSwNP和AERD患者中识别未诊断的AERD患者 患有哮喘。我们将确认或排除AERD的诊断与阿司匹林挑战，以验证MP测试。 这些研究将进一步加深对CRS和AERD发病机制的认识，并为我们带来 更接近新的诊断和治疗干预措施。

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.