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: 10116175
• 负责人: Matthew Camiolo
• 金额: $ 5.21万
• 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/10116175
• 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; 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; 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; 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; p38 Mitogen Activated Protein Kinase; pathogen; patient subsets; programs; pulmonary function; 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： 哮喘是一种常见病，全世界有超过3亿人受到影响。尽管控制得很好 在大多数情况下，一些人经历了对标准治疗(如皮质类固醇)难以治愈的疾病。近期 集中于了解这些严重哮喘(SA)患者的努力表明了本体的异质性。 重症哮喘研究计划(SARP)队列的先前工作将上皮基因签名与 临床表型。我们重新查看了这个数据集，确定了3个新的哮喘患者组，其中之一 容纳了大多数SA病例，具有最差的肺功能和最严重的恶化历史。vbl.使用 在数据集的主要差异驱动因素中，我们发现白细胞介素18受体(IL18R1)的表达 与其他18个基因一起，能够识别有风险的哮喘患者。IL18R1以前曾被认为与哮喘有关 全基因组关联研究。其配体IL-18可能是Th1和Th2的辅因子 发炎。哮喘患者的血清和痰中检测到高水平的IL-18，包括那些 致命的哮喘。重要的是，IL-18和IL18R1在其他炎症性疾病中调节上皮屏障功能 条件。我们已经证实，与健康人相比，SA患者IL18R1在蛋白水平上有所增加 对照组(HC)，使用非常严重的哮喘患者的肺活检。这些SA患者还增加了 编号IL-18+细胞，IL18R1的同源配体。IL18刺激的下游靶点，包括 活化磷酸化形式的JNK1和c-jun在SA呼吸道上皮细胞中表达增加。SA患者也 粘附性连接紊乱，提示上皮屏障完整性受损。 基于这一背景和我们的初步数据，我们假设IL-18途径通过 IL18R1在支气管上皮的表达增加，促进屏障功能失调，从而促进 SA病理生物学通过增加免疫细胞对外来抗原的获得性。我们提出2个目标来解决这个问题： 目的1.我们将询问现有的组织库以进行IL18R1的定量分析，并将其与 这些患者的组织和体液中的其配体水平。这些数据将被证实为 相关的信号通路和紧密/粘连的连接完整性。 目的2.我们将使用经支气管镜下从HCS和哮喘患者那里获得的原代人呼吸道上皮细胞 患者机械地确认和扩展来自目标1的结果。这将包括验证EX 活体结果以及屏障功能的量化。 这些数据将确定IL18在重症哮喘发病机制中的新作用，并开辟新的途径 用于治疗这种疑难且昂贵的疾病。该项目的活动将为PI提供有价值的 原代人呼吸道上皮细胞的获取、培养和操作经验 通过整合计算生物学和免疫学的研究，为未来的活体工作奠定基础。