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）： 哮喘是一种非常常见的疾病，在其一生中影响了6.4%的美国成年人1。确定哮喘发生和进展背后的分子机制将导致新的治疗靶点，更具体的治疗方案，以及更好地控制疾病。哮喘是一种炎症性疾病，其特征是平滑肌肥大、气道阻塞和粘液产生增加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起作用，然而疾病背后的分子机制尚未阐明。低氧诱导因子-11（HIF-11）是一种转录因子，最近被鉴定为髓样细胞炎症能力的主要调节因子4; 5。HIF-11介导炎症中巨噬细胞和中性粒细胞的许多关键功能4; 6.因为它存在于所有哺乳动物细胞中，调节超过100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调节哮喘中的过敏性炎症。 哮喘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，气道炎症的发展是对过敏原再暴露的反应。本研究的目的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，并探讨HIF-11发挥作用的分子机制。我假设（a）缺氧诱导因子-11是过敏性炎症分子通路的关键调节因子;（B）树突状细胞中的HIF-11在过敏性气道反应的启动中起重要作用;（c）嗜酸性粒细胞中的HIF-11在气道炎症的持续和进展中起作用。我将探讨白细胞中HIF-11水平的遗传操作如何影响哮喘易感性，并检查是否可以有效地抑制HIF-11的治疗策略可以减少气道炎症和反应性。 我们将在小鼠中利用卵清蛋白（OVA）过敏性炎症性气道疾病模型。将通过IgE、组胺、IL-4和IL-13水平、气道阻力、炎性肺浸润和气道重塑评价小鼠的过敏反应程度。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，细胞来源于小鼠骨髓。这些研究将采用迁移试验、FACS分析、ELISA、Western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。独特的小鼠基因试剂，其中HIF-11水平已显着降低或组成性增加白细胞谱系，可供我明确测试这些假设在体内和体外详细的，机械的方式。除了这些强大的遗传方法，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快速减少，其方式在停药后可迅速逆转。因此，如果严重哮喘患者长期接受HIF-11抑制剂治疗并发生感染，则可以暂时停用HIF-11拮抗剂以快速恢复正常白细胞功能。以及其他慢性炎性疾病（类风湿性关节炎、狼疮等），我推测抑制HIF-11可能为哮喘的治疗提供一个令人兴奋的策略。