Proteomic networks of MUC5B infectious/inflammatory induction in Otitis Media

中耳炎中 MUC5B 感染/炎症诱导的蛋白质组网络

• 批准号: 8276560
• 负责人: Diego Preciado
• 金额: $ 45.71万
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-15 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8276560
• 关键词: Abate; Acute; Amino Acids; Auditory; Bacterial Infections; Bioinformatics; Cell Culture Techniques; Cells; Child; Childhood; Chronic; Complement; Core Facility; Cytolysis; Data; Development; Disease; Dose; EMSA; Ear; Epithelial; Epithelium; Event; Exposure to; Fluids and Secretions; Functional disorder; Funding; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Generations; Haemophilus influenzae; Health Care Costs; Human; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Injury; Lead; Life; MUC5B gene; Mass Spectrum Analysis; Mediator of activation protein; Medical; Metaplasia; Modeling; Molecular; Molecular Target; Mucins; Mucous Membrane; Mucous body substance; Mus; National Institute on Deafness and Other Communication Disorders; Operative Surgical Procedures; Otitis Media; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Physicians; Play; Process; Proteins; Proteomics; Public Health; Publishing; RNA Interference; Research; Research Infrastructure; Series; Signal Transduction; Squamous Epithelium; Stable Isotope Labeling; Stimulus; Testing; Thick; Time; Treatment Failure; Tube; Tympanostomy; United States National Institutes of Health; Up-Regulation; Visit; airway epithelium; base; cytokine; ear infection; in vivo; interdisciplinary approach; knock-down; mRNA Expression; middle ear; middle ear disorder; mouse model; mucoid; novel; pathogen; programs; promoter; response; therapeutic target; transcription factor; treatment strategy

DESCRIPTION (provided by applicant: This application is a response to The National Institute on Deafness and Other Communication Disorders (NIDCD) issued PA-09-228 encouraging R01 applications that focus on Proteomics in Auditory Developmental and Disease Processes. Otitis media (OM), the most prevalent chronic childhood disorder, is associated with staggering public health care costs. It is a disease of the middle ear space characterized by acute infectious injury and inflammation, acute OM (AOM), progressing to chronic epithelial mucoid fluid secretion, i.e. chronic OM (COM). The OM continuum from acute to chronic is triggered by infectious or noxious stimuli that result in thickening of middle ear epithelia (MEE) and leads to self-sustaining chronic inflammation and mucous hypersecretion. This in turn often leads to medical treatment failure, and the placement of surgical tympanostomy tubes - the most common surgical procedure of children. In published and preliminary data, we have begun to characterize the molecular progression of OM events. We have recently shown that the hyperviscous mucus in COM is characterized by an overabundance of MUC5B mucin, a major airway secreted mucin. Through a gene expression profiling approach in a mouse model of acute OM based on middle ear inoculation of Non-typeable Hemophilus Influenza (NTHi) (currently the most common human AOM pathogen), we identified the pro-inflammatory cytokine Cxcl2 as markedly and acutely upregulated. Additionally, we observed that the middle ear epithelium chronically trans-differentiated into a thicker, pseudostratified state after repeatd NTHi lysate exposure. Here, we propose an interdisciplinary approach to understanding, and then intervening in, the molecular pathobiology of COM. We hypothesize that AOM, triggered by an inflammatory event (NTHi), activates expression of Cxcl2 and downstream expression of Muc5b/MUC5B, resulting in progression to chronic otitis media. We first aim to use a proteomic and cytokine secretome profiling approach to determine the in vitro effects of NTHi on a middle ear epithelial pro- inflammatory response, gaining important information that would complement our preliminary in vivo data. Secondly, we will analyze the effects of key inflammatory mediators identified in Aim 1 (predictably Cxcl2 and/or NF?B) on the expression of Muc5b. Finally, we will then aim to intervene at key molecular steps in the process (predictably Cxcl2, and/or NF?B) to halt progression to COM and middle ear Muc5b over-expression. The combination of our findings should result in a better understanding of pathways in OM pathophysiology and are likely to reveal novel molecular therapeutic targets that when modulated may help abate progression of AOM to COM. PUBLIC HEALTH RELEVANCE: Otitis Media (OM) is one of the most frequent disorders in children requiring physician visits. It is a disease of the middle ear space characterized by acute infectious injury and inflammation, acute OM (AOM), progressing to chronic epithelial mucoid fluid secretion, i.e. chronic OM (COM). In this application, using a proteomic profiling approach, we aim to interrogate how pathologically relevant acute infectious stimuli result in a cascade of inflammatory mediator up- regulation which in turn ultimately lead to middle ear epithelial metaplasia and inappropriate over-expression of mucins. Currently no medications exist to treat COM effectively. An understanding of the molecular mechanisms behind the progression of acute middle ear infection to chronic OM may radically change the way the disease is treated, especially if novel molecular targets are identified. Continuation Page

描述（由申请人提供：本申请是对国家耳聋和其他沟通障碍研究所 (NIDCD) 发布的 PA-09-228 鼓励 R01 应用的回应，该申请重点关注听觉发育和疾病过程中的蛋白质组学。中耳炎 (OM)，最常见的慢性儿童疾病，与惊人的公共医疗保健费用相关。它是一种以急性感染性损伤为特征的中耳疾病。 炎症，急性 OM (AOM)，进展为慢性上皮粘液分泌，即慢性 OM (COM)。从急性到慢性的 OM 连续体是由感染性或有害刺激触发的，导致中耳上皮 (MEE) 增厚，并导致自我维持的慢性炎症和粘液分泌过多。这往往会导致医疗失败，并导致放置外科鼓膜造口管——这是儿童最常见的外科手术。在已发表的初步数据中，我们已经开始描述 OM 事件的分子进展特征。我们最近发现 COM 中的高粘稠粘液的特点是 MUC5B 粘蛋白（一种主要气道分泌的粘蛋白）过多。通过在基于中耳接种不可分型嗜血杆菌流感 (NTHi)（目前最常见的人类 AOM 病原体）的急性 OM 小鼠模型中的基因表达谱分析方法，我们发现促炎细胞因子 Cxcl2 显着且急剧上调。此外，我们观察到，在反复接触 NTHi 裂解物后，中耳上皮长期转分化为较厚的假复层状态。在这里，我们提出了一种跨学科的方法来理解并干预 COM 的分子病理学。我们假设由炎症事件 (NTHi) 触发的 AOM 激活 Cxcl2 的表达和 Muc5b/MUC5B 的下游表达，导致进展为慢性中耳炎。我们首先旨在使用蛋白质组和细胞因子分泌组分析方法来确定 NTHi 对中耳上皮促炎症反应的体外影响，获得重要信息来补充我们的初步体内数据。其次，我们将分析目标 1 中确定的关键炎症介质（预计是 Cxcl2 和/或 NF?B）对 Muc5b 表达的影响。最后，我们的目标是干预该过程中的关键分子步骤（预计是 Cxcl2 和/或 NF?B），以阻止 COM 和中耳 Muc5b 过度表达的进展。我们的研究结果相结合应该可以更好地理解 OM 病理生理学途径，并可能揭示新的分子治疗靶点，这些靶点在调节时可能有助于减轻 AOM 向 COM 的进展。 公共卫生相关性：中耳炎 (OM) 是儿童最常见的疾病之一，需要就诊。这是一种中耳间隙疾病，其特征为急性 感染性损伤和炎症，急性 OM (AOM)，进展为慢性上皮粘液分泌，即慢性 OM (COM)。在本申请中，使用蛋白质组学分析方法，我们的目的是探讨病理相关的急性感染刺激如何导致炎症介质上调级联，进而最终导致中耳上皮化生和粘蛋白的不适当过度表达。目前尚无药物可有效治疗 COM。了解急性中耳感染发展为慢性 OM 背后的分子机制可能会从根本上改变该疾病的治疗方式，特别是如果确定了新的分子靶标的话。续页