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：
8991672
负责人：
BRYAN P HURLEY
金额：
$ 43.93万
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 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8991672
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 Pseudomonas aeruginosa pneumonia 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 inflammatory lung disease 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.

描述（由申请人提供）：粘膜上皮表面在物理意义上都是外部威胁的保护障碍，也是免疫监视和编排的重要贡献。呼吸道中传染性炎症性粘膜疾病的标志之一是跨粘膜屏障大规模动员中性粒细胞进入气道管腔。与致病生物相遇触发了一个过程，中性粒细胞从血液迁移到空域，以直接通过吞噬和释放有害物质来直接面对粘膜入侵者。中性粒细胞的积累可能会变得过度过度，导致宿主肺组织损伤，就像涉及铜绿假单胞菌疾病中的铜绿和囊性纤维化的传染性过程一样。中性粒细胞从血液到气道募集的最后一步涉及在粘膜表面的上皮屏障上迁移。我们的研究揭示了这种现象是基础的分子机制。使用在可渗透的Transwell过滤器上生长的肺上皮细胞，我们对响应感染的中性粒细胞上皮迁移进行了建模。我们的研究表明，用铜绿假单胞菌治疗肺上皮单层会导致磷脂酶A2的激活，这会导致蛛网膜酸的释放。然后，通过12-脂氧酶 /肝素合成酶的作用将花生四烯酸转化为中性粒细胞化学吸引肝素A3。肝素A3在肺上皮单层的顶部释放，并指导上皮屏障的中性粒细胞。在当前的应用中，我们旨在通过鉴定特定的磷脂酶A2和12-脂氧酶 /肝素合酶的特定磷脂酶A2和12-脂肪氧酶A2和12-脂肪氧酶合成酶（S），负责在病原体感染后产生肝蛋白A3，以进一步探索小鼠和人肺上皮细胞中的肝素A3产生的机制。此外，我们建议建立原代小鼠和人肺上皮细胞以及具有缺陷的囊性囊性纤维化跨膜电导调节剂的肺上皮细胞的空气界面培养物。在这些空气模型中，将评估铜绿假单胞菌激发肝素A3产生和嗜中性上皮迁移的能力。最后，我们将使用铜绿粘膜表面在体内使用小鼠嗜铜假单胞菌模型在体内评估肝素A3作为在气道粘膜表面运行的中性粒细胞化学吸引者。多种干预措施以干扰肝素A3合成，稳定性和作用，并将与包括病理组织损伤的严重程度，12-脂氧酶的表达以及中性粒细胞在感染空间中的积累相关的结果。这些研究旨在进一步阐明这种新欣赏的二十烷酸中性粒细胞化学吸引剂称为肝素A3，该肝素A3在各种粘膜表面产生。对这种新颖的先天炎症途径的更好理解具有巨大的希望，可以揭示一类不同的未开发目标，以利用治疗方法以减轻破坏性的肺部炎症。