Matrix Metalloproteinase-19 as a Mediator of Airway Fibrosis in Obese Allergic Asthma

基质金属蛋白酶-19 作为肥胖过敏性哮喘气道纤维化的调节剂

基本信息

批准号：
10201483
负责人：
Jennifer L. Ingram
金额：
$ 20.13万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-24 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10201483
关键词：
Affect Allergens Allergic Animal Model Anti-Inflammatory Agents Asthma Autocrine Communication Basement membrane Biopsy Bronchoalveolar Lavage Fluid CRISPR/Cas technology Cell physiology Cells Cellular Structures Chronic Chronic Disease Clustered Regularly Interspaced Short Palindromic Repeats Cryopreservation Data Deposition Development Enzymes Epithelial Epithelial Cells Exhibits Exposure to Extracellular Matrix Extrinsic asthma Fibroblasts Fibrosis GLP-I receptor Gastrectomy Gene Expression Goals Growth Factor Hormones House Dust Mite Allergens Human Immune response Impairment Incidence Individual Inflammatory Inflammatory Response Insulin Interleukin-13 Intervention Knock-out Knowledge Leptin Literature Lung Matrix Metalloproteinases Measures Mediating Mediator of activation protein Metabolic Modeling Mus Obese Mice Obesity Operative Surgical Procedures Paracrine Communication Pathogenicity Pathologic Pathologic Processes Patients Peptide Hydrolases Pharmaceutical Preparations Phenotype Physiology Play Process Production Proteins Proteomics Pulmonary Fibrosis Receptor Signaling Research Role Serum Severities Signal Transduction Structure Structure of parenchyma of lung Symptoms System Testing Thinness Tissues adipokines airway epithelium airway inflammation airway obstruction airway remodeling asthmatic asthmatic patient bariatric surgery base chemokine cytokine diet-induced obesity experience glucagon-like peptide 1 glucose metabolism improved in vivo insulin secretion matrix metalloproteinase 19 migration mouse model novel obese patients obesity treatment patient population peptide hormone pulmonary function response screening targeted treatment therapeutic target therapy development wound healing

项目摘要

Nearly 40% of the asthma patient population is obese. These patients exhibit increased asthma symptoms and severity. Obesity is associated with inflammatory and metabolic changes that can contribute to asthma pathobiology. Specifically, increased levels of leptin, a pro-inflammatory adipokine, and decreased secretion of glucagon-like peptide 1 (GLP-1), a peptide hormone that regulates insulin production, may augment pathogenic processes in asthma by acting directly on airway structural cells. In allergic asthma, airway epithelial cells produce pro-fibrotic mediators that activate airway fibroblasts to secrete excess extracellular matrix (ECM). These processes contribute to airway remodeling, structural changes in asthma that can result in permanent airway obstruction. The mechanisms directing airway remodeling in obese asthma are poorly understood, but in allergic asthma, these processes are directed in part by the type 2 cytokine, interleukin-13 (IL-13). Matrix metalloproteinase-19 (MMP-19) is an allergen-activated protease produced by airway epithelial cells and fibroblasts that participates in normal processing of ECM. Our preliminary data suggest that GLP-1 stimulates, while IL-13 and leptin suppress, MMP-19 production by airway cells. Our hypothesis is that obesity- and allergen-induced suppression of MMP-19 production plays an important role in the development of airway fibrosis in allergic asthma. Furthermore, GLP-1 inhibits pro-fibrotic cellular functions and halts the progression of airway fibrosis in obese, allergic humans and mice by acting to enhance airway fibroblast and epithelial cell MMP-19 secretion. To test this hypothesis, we propose two Aims. In Aim 1, we will culture primary airway epithelial cells and fibroblasts from obese and lean allergic asthmatic and obese and lean non-asthmatic subjects. We will silence MMP19 expression in these cells using CRISPR-Cas9 and determine the role of MMP-19 in pro-fibrotic cellular functions in response to leptin, IL-13 and GLP-1 or a GLP-1 receptor antagonist. In Aim 2, diet-induced obese Mmp19-I- and Mmp19+I+ mice will be concurrently challenged with house dust mite allergen for 4 weeks before undergoing vertical sleeve gastrectomy to induce GLP-1 secretion. We will assess airway fibrosis in vivo and relate these findings to GLP-1, leptin and MMP-19 levels in lung tissue, serum and bronchoalveolar lavage fluid. We will evaluate ex vivo cultured mouse lung fibroblasts for MMP-19 and ECM production. In both aims, we will relate pathologic airway changes and ex vivo cellular responses to measures of airway physiology and lung function in order to predict airway remodeling. Successful completion of these Aims will not only increase our understanding of the mechanisms directing the pathobiology of obese allergic asthma, but also will test specific interventions to treat obese asthma patients.

近 40% 的哮喘患者患有肥胖症。这些患者的哮喘症状加重症状和严重程度。肥胖与炎症和代谢变化有关，可能有助于哮喘病理学。具体来说，瘦素水平升高，促炎脂肪因子，并减少胰高血糖素样肽 1 (GLP-1)（一种肽）的分泌调节胰岛素产生的激素，可能通过作用增强哮喘的致病过程直接作用于气道结构细胞。在过敏性哮喘中，气道上皮细胞产生促纤维化介质，激活气道成纤维细胞分泌过量的细胞外物质矩阵（ECM）。这些过程有助于气道重塑和哮喘的结构变化，可能导致永久性气道阻塞。肥胖患者气道重塑的机制人们对哮喘知之甚少，但在过敏性哮喘中，这些过程部分是直接的由 2 型细胞因子白细胞介素 13 (IL-13) 引起。基质金属蛋白酶-19 (MMP-19) 是一种由气道上皮细胞和成纤维细胞产生的过敏原激活的蛋白酶参与 ECM 的正常处理。我们的初步数据表明 GLP-1 具有刺激作用，而 IL-13 和瘦素抑制气道细胞产生 MMP-19。我们的假设是肥胖和过敏原诱导的 MMP-19 产生抑制在疾病的发展中起着重要作用过敏性哮喘中的气道纤维化。此外，GLP-1 抑制促纤维化细胞功能，通过增强作用来阻止肥胖、过敏性人类和小鼠气道纤维化的进展气道成纤维细胞和上皮细胞分泌MMP-19。为了检验这个假设，我们提出了两个目标。在目标 1 中，我们将培养来自肥胖和瘦人的原代气道上皮细胞和成纤维细胞过敏性哮喘以及肥胖和瘦的非哮喘受试者。我们将沉默 MMP19 的表达这些细胞使用 CRISPR-Cas9 并确定 MMP-19 在促纤维化细胞功能中的作用响应瘦素、IL-13 和 GLP-1 或 GLP-1 受体拮抗剂。在目标 2 中，饮食引起的肥胖 Mmp19-I- 和 Mmp19+I+ 小鼠将同时接受屋尘螨过敏原攻击 4 次在进行垂直袖状胃切除术前几周诱导 GLP-1 分泌。我们将评估体内气道纤维化，并将这些发现与肺部 GLP-1、瘦素和 MMP-19 水平联系起来组织、血清和支气管肺泡灌洗液。我们将评估离体培养的小鼠肺用于 MMP-19 和 ECM 生产的成纤维细胞。在这两个目标中，我们都将病理性气道变化联系起来以及离体细胞对气道生理学和肺功能测量的反应，以便预测气道重塑。成功完成这些目标不仅会增加我们的了解指导肥胖过敏性哮喘病理学的机制，也将测试治疗肥胖哮喘患者的具体干预措施。