IL-18 Signaling and Lung Epithelial Cell Barrier Integrity in Severe Asthma

严重哮喘中的 IL-18 信号传导和肺上皮细胞屏障完整性

• 批准号: 9758736
• 负责人: Matthew Camiolo
• 金额: $ 7.22万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9758736
• 关键词: Address; Adherens Junction; Adrenal Cortex Hormones; Affect; Air; Allergens; Antigens; Asthma; Automobile Driving; Biological Assay; Biological Markers; Biology; Biopsy; Biopsy Specimen; Bronchoalveolar Lavage; Cells; Collection; Computational Biology; Cost of Illness; Data; Data Set; Disease; E-Cadherin; Electrical Resistance; Enzyme-Linked Immunosorbent Assay; Epithelial; Epithelial Cells; Epithelium; Event; Exhibits; Exposure to; Foundations; Functional disorder; Future; Genes; Heterogeneity; Housing; Human; IL18 gene; Immune; Immunofluorescence Microscopy; Immunohistochemistry; Immunology; Impairment; In Vitro; Inflammation; Inflammatory; Interleukin-18; JUN gene; Ligands; Link; Liquid substance; Lung; MAP Kinase Gene; MAPK14 gene; MAPK8 gene; Machine Learning; Measures; Mucous Membrane; Ontology; Pathogenesis; Pathway interactions; Patients; Permeability; Phenotype; Play; Positioning Attribute; Proteins; Pulmonary function tests; Quality of life; Recording of previous events; Refractory; Research; Research Personnel; Resources; Respiratory physiology; Risk; Role; Serum; Severity of illness; Signal Pathway; Signal Transduction; Source; Specimen; Sputum; Stains; Structure; Techniques; Tight Junctions; Tissue Banks; Tissues; Transcript; Transcription Factor AP-1; Treatment Cost; Universities; Validation; Variant; Western Blotting; Work; airway epithelium; asthma exacerbation; asthmatic; asthmatic airway; asthmatic patient; base; bronchial epithelium; clinical phenotype; cofactor; cohort; cytokine; experience; genetic signature; genome wide association study; in vivo; interleukin-18 receptor; novel; occludin; pathogen; patient subsets; programs; response; standard care; success; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT: Asthma is a common disease, affecting more than 300 million people worldwide. Though well controlled in most, some experience disease that is refractory to standard treatments such as corticosteroids. Recent efforts focused on understanding these severe asthma (SA) patients suggests ontological heterogeneity. Previous work from the Severe Asthma Research Program (SARP) cohort linked epithelial gene signature to clinical phenotype. We revisited this data set and identified 3 novel groups of asthmatics, one of which housing the majority of SA cases and having the worst lung function and greatest exacerbation history. Using principal drivers of variance in the data set, we found that expression of the interleukin 18 receptor (IL18R1) along with 18 other genes was able identify at-risk asthmatics. IL18R1 has been previously linked to asthma in genome-wide association studies. Its ligand, interleukin-18 (IL-18), may be a cofactor for both Th1 and Th2 inflammation. High IL-18 levels have been detected in serum and sputum of asthmatics including in those with fatal asthma. Importantly, IL-18 and IL18R1 regulate epithelial barrier function in other inflammatory conditions. We have confirmed an increase in IL18R1 at the protein level in SA patients compared to healthy controls (HC) using lung biopsies from very severe asthma cases. These SA patients also harbor increased numbers IL-18+ cells, the cognate ligand for IL18R1. Downstream targets of IL18 stimulation, including the active phosphorylated forms of JNK1 and c-Jun, show increase in SA airway epithelium. SA patients also show disorganization of adherens junctions, suggesting breakdown in epithelial barrier integrity. Based on this background and our preliminary data, we hypothesize that IL-18 pathway activation, via elevated IL18R1 expression in bronchial epithelium, promotes dysregulated barrier function that contributes to SA pathobiology by increasing immune cell access to foreign antigens. We propose 2 aims to address this: Aim 1. We will interrogate pre-existing tissue banks for quantitative analysis of IL18R1 and compare that with levels of its ligand in tissue and fluids from these patients. These data will be corroborated with activity of associated signaling pathways and tight/adherens junction integrity. Aim 2. We will us primary human airway epithelial cells, obtained bronchoscopically from HCs and asthmatic patients to mechanistically confirm and expand upon the results from Aim 1. This will include validation of ex vivo results as well as quantification of barrier function. These data will identify a novel role for IL18 in severe asthma pathogenesis and open up new avenues for treatment of this difficult and costly disease. The activities of the project will provide the PI with valuable experience in obtaining, culturing and manipulating primary human airway epithelial cells and lay the foundation for future in vivo work by integrating study in computational biology and immunology.

项目摘要/摘要： 哮喘是一种常见疾病，影响了全球超过3亿人。虽然控制得很好 在大多数情况下，某些经历了对标准治疗（例如皮质类固醇）难治性的疾病。最近的 专注于理解这些严重哮喘（SA）患者的努力表明本体论异质性。 严重哮喘研究计划（SARP）队列的先前工作将上皮基因签名与 临床表型。我们重新审视了这些数据集并确定了3种新型哮喘患者，其中之一 拥有大多数SA病例，具有最差的肺功能和最大的恶化史。使用 数据集中的主要方差驱动因素，我们发现白介素18受体的表达（IL18R1）的表达 除其他18个基因外，还可以识别出危险的哮喘患者。 IL18R1先前已与哮喘有关 全基因组关联研究。它的配体白介素18（IL-18）可能是Th1和Th2的辅助因子 炎。在哮喘患者的血清和痰中已经检测到高IL-18水平 致命哮喘。重要的是，IL-18和IL18R1调节其他炎症的上皮屏障功能 状况。我们已经证实，与健康相比，SA患者的蛋白质水平的IL18R1增加了 对照（HC）使用来自非常严重的哮喘病例的肺活检。这些SA患者还增加了 数字IL-18+细胞，IL18R1的同源配体。 IL18刺激的下游目标，包括 JNK1和C-JUN的主动磷酸化形式显示SA气道上皮的增加。 SA患者也是 显示粘附连接的混乱，表明上皮屏障完整性崩溃。 基于此背景和我们的初步数据，我们假设IL-18途径激活 支气管上皮中的IL18R1表达升高，促进了有助于 通过增加免疫细胞对外抗原的访问，SA病理生物学。我们提出2个目的是解决这一点： 目标1。我们将询问现有的组织库以进行定量分析IL18R1，并将其与 其在组织中的配体水平和这些患者的液体。这些数据将通过 相关的信号通路和紧密/粘附连接完整性。 AIM 2。我们将从HCS和哮喘中获得支气管镜的原发性人类气道上皮细胞 患者可以机械地确认并扩展AIM 1的结果。这将包括对EX的验证 体内结果以及屏障功能的定量。 这些数据将确定IL18在严重哮喘发病机理中的新作用，并打开新的途径 治疗这种困难且昂贵的疾病。该项目的活动将为PI提供宝贵的 在获得，培养和操纵人类气道上皮细胞并铺设的经验 通过整合计算生物学和免疫学研究的研究基础。