Genetic and Pharmacologic Evaluation of the Role of HIF-1a in asthma

HIF-1a 在哮喘中作用的遗传学和药理学评价

• 批准号: 8402123
• 负责人: Laura Elise Crotty Alexander
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8402123
• 关键词: Accident and Emergency department; Acute; Adult; Affect; Airway Resistance; Allergens; Allergic; Allergic inflammation; American; Animal Experiments; Antigen Presentation; Antigens; Asthma; Bone Marrow; Cells; Chronic; Complex; Cytoplasmic Granules; Data; Dendritic Cells; Development; Disease; Effector Cell; Evaluation; Extrinsic asthma; Family; Genes; Genetic; Goals; Goblet Cells; HIF1A gene; Half-Life; Healthcare; Histamine; Hospitalization; Hyperplasia; Hypersensitivity; Hypertrophy; Hypoxia Inducible Factor; IgE; Immunosuppressive Agents; In Vitro; Incidence; Infection; Inflammation; Inflammatory; Inflammatory Response; Interleukin-13; Interleukin-4; Laboratories; Lead; Leukocytes; Literature; Lung; Lupus; Lymphocyte; Mammalian Cell; Mediating; Medical; Mentors; Migration Assay; Military Personnel; Modeling; Molecular; Mucous body substance; Mus; Myelogenous; NF-kappa B; Navy Personnel; Ovalbumin; Pathogenesis; Pathway interactions; Peptides; Phagocytosis; Pharmaceutical Preparations; Phenotype; Phosphotransferases; Play; Population; Predisposition; Process; Production; Proteins; Reagent; Research; Rheumatoid Arthritis; Role; Secondary to; Series; Serum; Signal Transduction; Smooth Muscle; Stimulus; Structure of parenchyma of lung; System; Testing; Therapeutic; Time; Tissues; Veterans; Visit; Western Blotting; Work; YC-1; airway inflammation; airway obstruction; airway remodeling; allergic response; antigen challenge; asthmatic patient; cytokine; cytotoxic; design; disorder control; eosinophil; experience; extracellular; genetic manipulation; health care quality; human MAPK14 protein; hypoxia inducible factor 1; in vivo; inhibitor/antagonist; intraperitoneal; lipid mediator; macrophage; mast cell; member; migration; neutrophil; prevent; research study; response; small molecule; transcription factor; uptake

DESCRIPTION (provided by applicant): Asthma is an extremely common disease, affecting 6.4% of American adults over the course of their lives1. Determining the molecular mechanisms behind the development and progression of asthma will lead to new treatment targets, more specific treatment options, and better control of the disease. Asthma is an inflammatory disease characterized by hypertrophy of smooth muscle, airway obstruction, and increased mucous production3. Dendritic cells, eosinophils, and mast cells are known to play a role in the development and progression of asthma, however the molecular mechanisms behind the disease have not been elucidated. Hypoxia inducible factor-11 (HIF-11) is a transcription factor recently identified as a master regulator of the inflammatory capacity of myeloid cells4; 5. HIF-11 mediates many critical functions of macrophages and neutrophils in inflammation4; 6. Because it is found in all mammalian cells, regulates over 100 functional genes, and modulates NF-:B pathways that are known to be up regulated during allergic inflammation7; 8, I hypothesize that HIF-11 regulates allergic inflammation in asthma. The pathogenesis underlying the development of asthma can be broken down into two components. First, an allergic response to a specific allergen is initiated. Second, airway inflammation develops in response to allergen re-exposure. The goal of the work proposed here is to explore how the transcription factor Hypoxia Inducible Factor-11 (HIF-11) modulates the pathogenesis of asthma, and to probe mechanistically the molecular pathways through which HIF-11 acts in this capacity. I hypothesize that (a) Hypoxia Inducible Factor-11 is a critical regulator of molecular pathways involved in allergic inflammation; (b) HIF-11 in dendritic cells plays a significant role in the initiation of the allergic airway response; and (c) HIF-11 in eosinophils plays a role in the perpetuation and progression of airway inflammation. I will explore how genetic manipulation of HIF-11 levels in leukocytes influences asthma susceptibility, and examine whether a therapeutic strategy to pharmacologically inhibit HIF-11 can be effective in reducing airway inflammation and reactivity. We will utilize an ovalbumin (OVA) model of allergic inflammatory airways disease in mice. Mice will be evaluated for the magnitude of allergic response via IgE, histamine, IL-4, and IL-13 levels, airway resistance, inflammatory pulmonary infiltrates, and airway remodeling. Detailed, mechanistic studies of eosinophils and dendritic cells will be done in vitro, with cells derived from mouse bone marrow. These studies will utilize migration assays, FACS analysis, ELISAs, Western blots, and real-time quantitative PCR. In this proposal, I will determine whether HIF-11 initiates or sustains allergic inflammatory airway disease development in mice. Specifically, these studies will rigorously test whether HIF-11 regulates dendritic cells and eosinophil function in allergic inflammation leading to asthma. Unique mouse genetic reagents, in which HIF-11 levels have been markedly reduced or constitutively increased in leukocyte lineages, are available for me to definitively test these hypotheses in a detailed, mechanistic fashion in vivo and ex vivo. Beyond these powerful genetic approaches, I propose a parallel series of pharmacologic experiments utilizing small molecule inhibitors to globally suppress HIF-11. I hypothesize that because HIF-11 is regulated at the protein level with a half-life of seconds9, anti-HIF-11 immunosuppressant agents will lead to a rapid decrease in inflammation in a manner that is rapidly reversible upon discontinuation of the agent. Therefore, if severe asthmatic patients were treated chronically with a HIF-11 inhibitor and developed an infection, the HIF-11 antagonist could be temporarily stopped to provide a rapid return of normal leukocyte function. As well as other chronic inflammatory diseases (rheumatoid arthritis, lupus, etc.), I hypothesize that HIF-11 inhibition may offer an exciting strategy for the management of asthma.

描述（由申请人提供）： 哮喘是一种极其常见的疾病，影响 6.4% 的美国成年人一生1。确定哮喘发生和进展背后的分子机制将带来新的治疗目标、更具体的治疗选择以及更好的疾病控制。哮喘是一种炎症性疾病，其特征是平滑肌肥大、气道阻塞和粘液分泌增多3。已知树突状细胞、嗜酸性粒细胞和肥大细胞在哮喘的发生和进展中发挥作用，但该疾病背后的分子机制尚未阐明。缺氧诱导因子 11 (HIF-11) 是一种转录因子，最近被确定为骨髓细胞炎症能力的主要调节因子4； 5. HIF-11介导巨噬细胞和中性粒细胞在炎症中的许多关键功能4； 6. 因为它存在于所有哺乳动物细胞中，调节 100 多个功能基因，并调节已知在过敏性炎症期间上调的 NF-:B 通路7； 8、我假设 HIF-11 调节哮喘的过敏性炎症。 哮喘发生的发病机制可分为两个部分。首先，引发对特定过敏原的过敏反应。其次，气道炎症是因再次接触过敏原而发生的。本文提出的工作目标是探索转录因子缺氧诱导因子 11 (HIF-11) 如何调节哮喘的发病机制，并从机制上探讨 HIF-11 发挥这种作用的分子途径。我假设 (a) 缺氧诱导因子 11 是参与过敏性炎症的分子途径的关键调节因子； (b) 树突状细胞中的 HIF-11 在过敏性气道反应的启动中发挥重要作用； (c) 嗜酸性粒细胞中的 HIF-11 在气道炎症的持续和进展中发挥作用。我将探讨白细胞中 HIF-11 水平的基因操作如何影响哮喘易感性，并研究药物抑制 HIF-11 的治疗策略是否可以有效减少气道炎症和反应性。 我们将利用小鼠过敏性炎症性气道疾病的卵清蛋白 (OVA) 模型。将通过 IgE、组胺、IL-4 和 IL-13 水平、气道阻力、炎症性肺部浸润和气道重塑来评估小鼠过敏反应的程度。对嗜酸性粒细胞和树突状细胞的详细机制研究将在体外进行，细胞来自小鼠骨髓。这些研究将利用迁移测定、FACS 分析、ELISA、蛋白质印迹和实时定量 PCR。 在本提案中，我将确定 HIF-11 是否会引发或维持小鼠过敏性炎症性气道疾病的发展。具体来说，这些研究将严格测试 HIF-11 是否在导致哮喘的过敏性炎症中调节树突状细胞和嗜酸性粒细胞功能。独特的小鼠遗传试剂，其中白细胞谱系中的 HIF-11 水平显着降低或组成性增加，可供我在体内和体外以详细、机制的方式明确检验这些假设。除了这些强大的遗传方法之外，我还提出了一系列平行的药理学实验，利用小分子抑制剂来全面抑制 HIF-11。我推测，由于 HIF-11 在蛋白质水平上受到调节，半衰期为秒9，因此抗 HIF-11 免疫抑制剂将导致炎症迅速减轻，停药后可迅速逆转。因此，如果严重哮喘患者长期接受 HIF-11 抑制剂治疗并出现感染，可以暂时停用 HIF-11 拮抗剂，以快速恢复正常白细胞功能。与其他慢性炎症性疾病（类风湿性关节炎、狼疮等）一样，我推测 HIF-11 抑制可能为哮喘的治疗提供令人兴奋的策略。