Molecular and Cellular Dissection of miRNAs in Controlling Allergic Airway Inflammation in Mice

miRNA 在控制小鼠过敏性气道炎症中的分子和细胞解析

• 批准号: 10477187
• 负责人: DAVID B CORRY
• 金额: --
• 依托单位: MICHAEL E DEBAKEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10477187
• 关键词: Address; Affect; Airway Disease; Allergens; Allergic; Allergic Disease; Allergic inflammation; Antifungal Agents; Antigen-Presenting Cells; Antigens; Aspergillus niger; Asthma; C-Type Lectins; CD11c Antigens; CD4 Positive T Lymphocytes; Cells; Chronic; Complex; Dendritic Cells; Dendritic cell activation; Detection; Development; Disease; Dissection; Epithelial Cells; Exposure to; Extrinsic asthma; FOXO1A gene; Family; Fungal Spores; Gene Expression; Genetic; Goals; Human; ITGAX gene; IgE; Immune response; In Vitro; Inflammatory; Inhalation; Interleukin-10; Interleukin-13; Knockout Mice; Laboratories; Lung; Lymphocyte; Mediating; Methods; MicroRNAs; Molecular; Molecular Analysis; Molecular Genetics; Mus; Myeloid Cells; Pathogenesis; Pathologic; Pathway interactions; Patients; Pattern recognition receptor; Peptide Hydrolases; Production; Proteinase-Activated Receptors; Pulmonary Inflammation; Refractory; Regulation; Role; Severity of illness; Suggestion; Testing; Therapeutic; Transgenic Mice; Veterans; airway epithelium; airway hyperresponsiveness; airway inflammation; airway remodeling; allergic airway disease; allergic airway inflammation; asthmatic; asthmatic patient; base; cell type; chronic inflammatory lung disease; combat veteran; conditional knockout; cytokine; fungus; genetic element; genome-wide; global health; improved; in vivo; insight; member; mouse model; novel diagnostics; novel strategies; novel therapeutics; programs; response; specific biomarkers; standard care; success; tool; transcriptome

The long-term objective of our laboratory is to understand the underlying molecular and genetic causes for chronic allergic airway inflammation or asthma to improve therapeutic approaches. The specific objective of this application is to dissect microRNA (miRNA)-dependent inflammatory pathways in dendritic cells (DCs) and CD4+ T cells that modulate murine allergic airway disease to fungal allergens. Asthma is a major rising global health threat that disproportionately affects combat veterans. We showed that patients with asthma often have evidence of airway mycosis, defined as detection of fungi in airway secretions in association with specific biomarkers (e.g. IgE or TH2 cells) reactive to one or more fungi. Our laboratory and others have shown that fungal proteinases and fungal spores drive asthma-like airway disease in mice. Airway epithelial cells express pattern recognition receptors (PRRs) for foreign antigens (e.g. TLRs, C-type lectin, and protease-activated receptors) and important for conventional dendritic cell (cDC) activation and initiation of allergic CD4+ type 2 cell (TH2) and TH17 cell-dependent airway inflammation, IgE production, and airway hyperresponsiveness (AHR), collectively termed allergic airway disease. We and others have shown the requirement of TH2 cells and DC in the pathogenesis of allergic airway disease. We employed murine models of chronic allergic airway inflammation, and detected aberrant expression of let-7 miRNA family in lung, CD4+ T cells and CD11c+ DCs. Specifically, the expression of let-7b and let-7c (let-7bc-cluster) is reduced in sorted lung CD4+ T cells and enhanced in CD11c+ antigen presenting cells (APCs) of mice with chronic allergic airway disease. Moreover, mice that lack the let-7bc-cluster globally or specifically in CD4+ T cells or CD11c+ APCs show blunted TH2 responses and allergic disease. On the other hand enforced expression of let-7 specifically in CD4+ T cells enhances fungus-induced allergic airway disease. The let-7b and let-7c members only account for only 9% of total let-7 activity in lymphocytes and myeloid cells suggesting that let-7bc cluster critically determines gene expression of CD4+ T cells and DCs that drive allergic sensitization to Aspergillus niger. Our central hypothesis therefore states that the let-7 family modulates allergic airway disease severity and orchestrates activation of cDCs and TH2 responses. We will address this hypothesis through the following Specific Aims: 1) Elucidate the CD4+ T cell intrinsic requirement of the let-7bc-cluster in pathogenesis of allergic airway disease. Hypothesis: The let-7bc-cluster regulates target gene expression in CD4+ T cells and controls TH2 lung inflammation and allergic airway disease. 2) Determine the role of let-7bc-cluster in DC-mediated control of allergic airway inflammation. Hypothesis: Induction of let-7bc-cluster expression in asthmatic lung cDCs directs molecular programming of TH2 driven pulmonary allergic inflammation. Through these two aims, we will further define the mechanisms by which the let-7bc cluster controls the expression of allergic airway disease, providing novel diagnostic and therapeutic insight into asthma and related disorders.

我们实验室的长期目标是了解潜在的分子和遗传原因 改善慢性过敏性气道炎症或哮喘的治疗方法。具体目标 该应用旨在剖析树突状细胞 (DC) 中 microRNA (miRNA) 依赖性炎症通路 CD4+ T 细胞可调节小鼠对真菌过敏原的过敏性气道疾病。哮喘是全球主要的新兴疾病 健康威胁对退伍军人的影响尤为严重。我们发现哮喘患者经常有 气道真菌病的证据，定义为在气道分泌物中检测到与特定疾病相关的真菌 对一种或多种真菌有反应的生物标志物（例如 IgE 或 TH2 细胞）。我们的实验室和其他实验室已经表明 真菌蛋白酶和真菌孢子会导致小鼠出现哮喘样气道疾病。气道上皮细胞表达 外来抗原的模式识别受体 (PRR)（例如 TLR、C 型凝集素和蛋白酶激活 受体），对于常规树突状细胞 (cDC) 激活和过敏性 CD4+ 2 型的启动很重要 细胞 (TH2) 和 TH17 细胞依赖性气道炎症、IgE 产生和气道高反应性 (AHR)，统称为过敏性气道疾病。我们和其他人已经证明了 TH2 细胞的需求 和 DC 在过敏性气道疾病发病机制中的作用。我们采用了慢性过敏性气道的小鼠模型 炎症，并检测到肺、CD4+ T 细胞和 CD11c+ DC 中 let-7 miRNA 家族的异常表达。 具体而言，在分选的肺 CD4+ T 细胞中，let-7b 和 let-7c (let-7bc-cluster) 的表达减少，并且 患有慢性过敏性气道疾病的小鼠的 CD11c+ 抗原呈递细胞 (APC) 增强。而且， 全局或特定于 CD4+ T 细胞或 CD11c+ APC 中缺乏 let-7bc 簇的小鼠显示 TH2 减弱 反应和过敏性疾病。另一方面，let-7 在 CD4+ T 细胞中特异性表达 增强真菌引起的过敏性气道疾病。 let-7b和let-7c成员仅占9% 淋巴细胞和骨髓细胞中总的let-7活性表明let-7bc簇关键地决定基因 CD4+ T 细胞和 DC 的表达驱动对黑曲霉的过敏致敏。我们的中心假设 因此，表明let-7家族调节过敏性气道疾病的严重程度并协调激活 cDC 和 TH2 反应。我们将通过以下具体目标来解决这一假设：1）阐明 过敏性气道疾病发病机制中 let-7bc 簇的 CD4+ T 细胞内在需求。 假设：let-7bc-cluster 调节 CD4+ T 细胞中的靶基因表达并控制 TH2 肺 炎症和过敏性气道疾病。 2) 确定let-7bc-cluster在DC介导的控制中的作用 过敏性气道炎症。假设：哮喘肺 cDC 中 let-7bc 簇表达的诱导指导 TH2 驱动的肺部过敏性炎症的分子编程。通过这两个目标，我们将进一步 定义let-7bc簇控制过敏性气道疾病表达的机制， 为哮喘及相关疾病提供新的诊断和治疗见解。