The Role of Hepoxilin A3 in Neutrophil Breach of the Infected Airway Mucosa

赫泊西林 A3 在中性粒细胞突破感染气道粘膜中的作用

基本信息

批准号：
8244864
负责人：
BRYAN P HURLEY
金额：
$ 44.1万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-08 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8244864
关键词：
AIDS/HIV problem Acute Age Air Animal Model Anti-Inflammatory Agents Anti-inflammatory Apical Arachidonate 12-Lipoxygenase Arachidonic Acids Automobile Driving Biological Models Blood Circulation CXC Chemokines Cell Culture Techniques Charge Child Chronic Complement 5a Complex Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Development Disease Eicosanoids Enzymes Epithelial Epithelial Cells Exhibits Failure Family Future Genes Health Human Immigration Immunologic Surveillance In Vitro Inborn Genetic Diseases Individual Infection Inflammation Inflammation Mediators Inflammatory Injury Intervention Investigation Knowledge Leukotriene B4 Leukotrienes Lipids Liquid substance Lung Lung Inflammation Lung diseases Malaria Malignant Neoplasms Membrane Modeling Molecular Mucous Membrane Mus Myocardial Infarction Neutrophil Infiltration Organism Outcome Pathology Pathway interactions Phospholipase Phospholipase A2 Phospholipids Play Pneumonia Process Production Prostaglandins Protein Isoforms Pseudomonas aeruginosa Publications Recruitment Activity Resolution Respiratory System Respiratory tract structure Role Severities Signal Transduction Site Staging Stroke Structure of parenchyma of lung Surface Therapeutic Tissues Work airway epithelium base design in vitro Model in vivo information gathering member migration monolayer mouse model neutrophil novel pathogen response therapeutic target

项目摘要

DESCRIPTION (provided by applicant): The mucosal epithelial surface serves as a protective barrier to external threats both in the physical sense and as a vital contributor to immune surveillance and orchestration. One of the hallmarks of infectious inflammatory mucosal disease in the respiratory tract is massive mobilization of neutrophils across the mucosal barrier into the airway lumen. Encounter with a pathogenic organism triggers a process whereby neutrophils migrate from the bloodstream to airspace in order to directly confront mucosal invaders through engulfment and release of noxious substances. Neutrophil accumulation can become over-exuberant resulting in host lung tissue damage as with the infectious process involving Pseudomonas aeruginosa in the diseases pneumonia and cystic fibrosis. The final step in neutrophil recruitment from the bloodstream to the airway involves migration across the epithelial barrier of the mucosal surface. Our studies have revealed a molecular mechanism that underlies this phenomenon. Using lung epithelial cells grown on permeable Transwell filters, we have modeled neutrophil trans-epithelial migration in response to infection. Our studies have demonstrated that treatment of lung epithelial monolayers with P. aeruginosa results in activation of phospholipase A2, which causes the release of arachidonic acid. Arachidonic acid is then converted to the neutrophil chemo-attractant hepoxilin A3 by the actions of 12-lipoxygenases / hepoxilin synthases. Hepoxilin A3 is released at the apical surface of lung epithelial monolayers and guides neutrophils across the epithelial barrier. In the current application, we aim to further explore the mechanism underlying hepoxilin A3 production in mouse and human lung epithelial cells by identifying both the specific phospholipase A2 and 12-lipoxygenase / hepoxilin synthase enzyme(s) responsible for generating hepoxilin A3 upon infection with pathogen. Furthermore, we propose to establish air-liquid interface cultures of primary mouse and human lung epithelial cells as well as lung epithelial cells with defective cystic fibrosis transmembrane conductance regulator. The ability of P. aeruginosa to instigate hepoxilin A3 production and neutrophil trans-epithelial migration will be assessed in these air-liquid models. Finally, we will evaluate hepoxilin A3 as a neutrophil chemo-attractant operating at the airway mucosal surface in vivo employing a mouse model of P. aeruginosa pneumonia. Multiple interventions to interfere with hepoxilin A3 synthesis, stability, and action will be utilized and correlated with outcomes that include severity of pathological tissue damage, expression of 12-lipoxygenases, and neutrophil accumulation in the infected airspace. These studies aim to further elucidate the role of this newly appreciated eicosanoid neutrophil chemo-attractant termed hepoxilin A3 that is produced at various mucosal surfaces. A better understanding of this novel innate inflammatory pathway holds tremendous promise towards unveiling a distinct class of unexplored targets to exploit therapeutically in order to alleviate destructive lung inflammation. PUBLIC HEALTH RELEVANCE: The objective of this application is to further investigate the hypothesis that hepoxilin A3 represents an important molecule that is critical for directing neutrophils to breach the mucosal epithelial barrier. Information gathered from this work will be instrumental in the development of a novel class of anti- inflammatory therapeutics that targets the destructive neutrophil driven tissue injury that occurs in a wide array of lung inflammatory diseases, both infectious and otherwise.

描述（由申请人提供）：粘膜上皮表面在物理意义上充当外部威胁的保护屏障，并且作为免疫监视和协调的重要贡献者。呼吸道传染性炎症粘膜疾病的标志之一是大量中性粒细胞穿过粘膜屏障进入气道腔。遇到病原生物会触发一个过程，中性粒细胞从血流迁移到空气空间，以便通过吞噬和释放有毒物质来直接对抗粘膜入侵者。中性粒细胞积聚可能变得过度旺盛，导致宿主肺组织损伤，就像肺炎和囊性纤维化疾病中涉及铜绿假单胞菌的感染过程一样。中性粒细胞从血流募集到气道的最后一步涉及迁移穿过粘膜表面的上皮屏障。我们的研究揭示了这种现象背后的分子机制。使用在可渗透的 Transwell 过滤器上生长的肺上皮细胞，我们模拟了中性粒细胞响应感染的跨上皮迁移。我们的研究表明，用铜绿假单胞菌处理肺上皮单层会导致磷脂酶 A2 的激活，从而导致花生四烯酸的释放。然后，花生四烯酸通过 12-脂氧合酶/Hepoxilin 合酶的作用转化为中性粒细胞趋化剂 Hepoxilin A3。 Hepoxilin A3 在肺上皮单层的顶端表面释放并引导中性粒细胞穿过上皮屏障。在当前的应用中，我们的目标是通过鉴定在病原体感染时负责生成Hepoxilin A3的特定磷脂酶A2和12-脂氧合酶/Hepoxilin合酶，进一步探索小鼠和人肺上皮细胞中Hepoxilin A3生成的机制。。此外，我们建议建立原代小鼠和人肺上皮细胞以及囊性纤维化跨膜电导调节器缺陷的肺上皮细胞的气液界面培养物。将在这些空气-液体模型中评估铜绿假单胞菌促进赫泊西林 A3 产生和中性粒细胞跨上皮迁移的能力。最后，我们将使用铜绿假单胞菌肺炎小鼠模型来评估赫泊西林 A3 作为中性粒细胞趋化剂在体内气道粘膜表面的作用。将利用多种干预措施来干扰赫泊西林 A3 的合成、稳定性和作用，并将其与结果相关联，包括病理组织损伤的严重程度、12-脂氧合酶的表达以及受感染空域中中性粒细胞的积累。这些研究旨在进一步阐明这种新近被认可的类花生酸中性粒细胞化学引诱剂（称为赫泊西林 A3）的作用，它是在各种粘膜表面产生的。更好地了解这种新的先天炎症途径，为揭示一类独特的未探索靶标提供了巨大的希望，这些靶标可用于治疗，以减轻破坏性肺部炎症。公共健康相关性：本申请的目的是进一步研究以下假设：赫泊西林 A3 是一种重要分子，对于引导中性粒细胞突破粘膜上皮屏障至关重要。从这项工作中收集的信息将有助于开发一类新型抗炎疗法，该疗法针对破坏性中性粒细胞驱动的组织损伤，这种损伤发生在多种肺部炎症性疾病中，包括传染性和其他疾病。