Determining the Role of the Upper and Lower Airway Microbiota as Drivers of Concomitant Inflammatory Responses in patients with Chronic Rhinosinusitis and Asthma

123 四五六 七百八十九 1101 1123 1145 1167 1189 2201 2223 2245 2267 2289 摘要 尽管人们越来越努力地了解微生物区系在呼吸道疾病中的作用，但与 微生物群落对慢性炎症的生态失调仍然难以捉摸。慢性鼻窦炎(CRS)是一种 美国的重大健康问题，每年造成高达650亿美元的损失，影响高达16%的 美国人口。CRS患者经常出现肺部合并症，这表明有 导致上呼吸道和下呼吸道炎症的共同潜在病理生理机制， 被称为统一航空公司的概念。在这项建议中，我们旨在了解上级和下级的作用 呼吸道微生物区系在驱动CRS和哮喘患者并发免疫反应中的作用。哮喘是一种 与CRS有很强的临床和流行病学联系的常见肺部疾病：20%- 60%的CRS鼻息肉患者有哮喘。据报道，在 然而，CRS患者的鼻窦和哮喘患者的肺中，微生物群或宿主免疫反应 这些地点还没有同时进行审查。这项提案的目标是确定存在一个 通过对细菌和真菌微生物组进行测序来确定上-下呼吸道轴，描绘宿主免疫状况 响应，并通过旨在鉴定混合物种培养中细菌代谢物的体外实验 会导致2型炎症。支持统一呼吸道的概念，我们的初步数据显示 慢性阻塞性肺综合征合并哮喘及合并2型患者上、下呼吸道微生物群落研究 炎症在整个呼吸道部位都有。为了扩展这些研究，我们将使用集成的多组学 研究微生物组结构和功能的基本基础的方法 CRS患者匹配的上、下呼吸道标本(有或无)的免疫应答 哮喘)和我们在过去四年中储存的健康个体。从这些分析中，我们将会 能够推断导致并发上下部慢性炎症的生态关系 呼吸道疾病。在第二个目标中，我们将确定致病微生物群落是否以及如何驱动 或在体外加剧呼吸道炎症反应。我们将描述CRS/哮喘的代谢组- 并在混合菌种生物膜培养中确认这些代谢物是微生物来源。要确定 与宿主的相互作用，我们将致敏与CRS-哮喘相关的外周血树突状细胞(DC) 代谢产物，然后与原始T细胞共培养树突状细胞，并量化T细胞分化。这项研究将有助于 有助于我们对统一性呼吸道疾病上下气道轴的认识，并最终导致 治疗学的目标是操纵上呼吸道微生物群来治疗并发的鼻窦和肺 疾病。

123 456 789 1101 1123 1145 1167 1189 2201 2223 2245 2267 2289 ABSTRACT Despite growing efforts to understand the role of the microbiota in airway disease, mechanisms that link microbial community dysbiosis to chronic inflammation remain elusive. Chronic rhinosinusitis (CRS) is a significant health problem in the United States, costing up to $65 billion each year and affecting up to 16% of the US population. CRS patients frequently present with pulmonary comorbidities suggesting that there are common underlying pathophysiologic mechanisms that contribute to upper and lower airway inflammation, known as the unified airways concept. In this proposal, we aim to understand the role of the upper and lower airway microbiota in driving concurrent immune responses in patients with CRS and asthma. Asthma is a common pulmonary disease that has strong clinical and epidemiological associations with CRS: Between 20%- 60% of CRS patients with nasal polyps have asthma. Dysbiotic microbial communities have been reported in sinuses of CRS patients and in the lungs of asthmatics, however, the microbiome or host immune response of these sites have not been examined in parallel. The goal of this proposal is to establish the existence of an upper-lower airway axis by sequencing the bacterial and fungal microbiome, profiling the host immune response, and through in vitro experiments that aim to identify bacterial metabolites in mixed-species culture that drive type-2 inflammation. Supporting the concept of a unified airway, our preliminary data show dysbiotic microbial communities in the upper and lower airways of CRS patients with asthma and concurrent type-2 inflammation shared across the airway sites. To expand these studies, we will use an integrated multi-omics approach to investigate the fundamental basis of microbiome structure and function in the context of the immune response of patient-matched upper and lower airway samples from CRS patients (with or without asthma) and healthy individuals that we have banked over the past four years. From these analyses, we will be able to infer the ecological relationships that contribute to chronic inflammation in concurrent upper and lower airway disease. In the second aim, we will determine whether and how pathogenic microbial communities drive or exacerbate airway inflammatory responses in vitro. We will characterize the metabolome in CRS/Asthma- patients and confirm these metabolites are of microbial origin in mixed species biofilm culture. To determine interaction with the host, we will sensitize peripheral blood dendritic cells (DCs) with CRS-Asthma associated metabolites, then co-culture DCs with naïve T cells and quantify T cell differentiation. This study will contribute to our understanding of the upper-lower airway axis in unified airway disease and will ultimately lead to therapeutics aimed a manipulating the upper airway microbiome for treating concurrent sinus and lung disease.