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: 10012111
• 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/10012111
• 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; Lung Inflammation; 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; 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; combat; conditional knockout; cytokine; fungus; genetic element; genome-wide; global health; improved; in vivo; inflammatory lung disease; 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其潜在的分子和遗传原因。 慢性过敏性呼吸道炎症或哮喘，以改进治疗方法。的具体目标 本应用旨在剖析树突状细胞(DC)中依赖于microRNA(MiRNA)的炎症途径。 调节小鼠对真菌变应原过敏的呼吸道疾病的CD4+T细胞。哮喘是全球发病率上升的主要原因 对退伍军人影响不成比例的健康威胁。我们发现哮喘患者通常有 呼吸道真菌病的证据，定义为检测与特定疾病相关的呼吸道分泌物中的真菌 对一种或多种真菌有反应的生物标志物(如IgE或TH2细胞)。我们的实验室和其他实验室已经证明 真菌蛋白水解酶和真菌孢子会导致小鼠的哮喘样呼吸道疾病。呼吸道上皮细胞表达 外来抗原的模式识别受体(如TLRs、C型凝集素和蛋白酶激活的 受体)，对传统的树突状细胞(CDC)激活和启动变态反应性CD4+2型很重要 细胞(TH2)和TH17细胞依赖的气道炎症、IgE产生和气道高反应性 (AHR)，统称为过敏性呼吸道疾病。我们和其他人已经证明了TH2细胞的需求 DC在过敏性呼吸道疾病发病机制中的作用。我们使用了慢性过敏性呼吸道的小鼠模型 肺组织、CD4+T细胞和CD11c+DC中检测到let-7miRNA家族的异常表达。 具体地说，在分离的肺CD4+T细胞中，let-7b和let-7c(let-7BC-group)的表达减少 慢性过敏性呼吸道疾病小鼠CD11c+抗原提呈细胞(APC)增强。此外， 在全球范围内或在CD4+T细胞或CD11c+APC中缺乏let-7BC-簇的小鼠表现为钝化的TH2 反应和过敏性疾病。另一方面，在CD4+T细胞中特异性表达let-7 增强真菌引起的过敏性呼吸道疾病。LET-7b和LET-7c成员仅占9% 淋巴细胞和髓系细胞中let-7的总活性表明let-7BC簇是决定基因的关键 对黑曲霉致敏的CD_4~+T细胞和DC的表达我们的中心假设 因此指出，let-7家族调节过敏性呼吸道疾病的严重程度，并协调激活 CDC和TH2应答。我们将通过以下具体目标来解决这一假设：1)阐明 过敏性呼吸道疾病发病机制中let-7BC-簇对CD4+T细胞的内在要求 假设：let-7BC簇调节CD4+T细胞的靶基因表达并控制TH2肺 炎症和过敏性呼吸道疾病。2)确定let-7BC-CLUSTER在DC介导的调控中的作用 过敏性呼吸道炎症。假设：诱导哮喘患者肺CDC表达let-7BC簇 TH2驱动的肺变态反应性炎症的分子程序设计。通过这两个目标，我们将进一步 确定let-7BC簇控制过敏性呼吸道疾病表达的机制， 为哮喘和相关疾病提供新的诊断和治疗见解。