Regulation of Allergen-Induced Airway Pathophysiology by Versican

Versican 对过敏原诱导的气道病理生理学的调节

• 批准号: 10327691
• 负责人: Charles Wayne Frevert
• 金额: $ 76.27万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-02 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10327691
• 关键词: Address; Adhesions; Agonist; Allergens; Anti-Inflammatory Agents; Asthma; Binding; Biopsy; Cell Culture Techniques; Cell physiology; Cells; Characteristics; Chronic; Data; Development; Disease; Environment; Epithelial Cells; Exposure to; Extracellular Matrix; Fibroblasts; Functional disorder; Genes; Glycosaminoglycans; Homeostasis; House Dust Mite Allergens; Human; Immune response; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Interferon Type I; Knowledge; Leukocytes; Lung; Lung diseases; Measures; Modeling; Modification; Morphogenesis; Mouse Strains; Mus; Myelogenous; Myeloid Cells; Organ; Pathway interactions; Patients; Pattern recognition receptor; Phenotype; Poly C; Poly I-C; Production; Property; Protein Isoforms; Publishing; Pulmonary Inflammation; Pyroglyphidae; Regulation; Role; Sampling; Side; Signal Pathway; Source; Stromal Cells; Structure; TCF Transcription Factor; Tamoxifen; Testing; Therapeutic; Tissues; Toll-like receptors; Translating; Work; airway epithelium; airway hyperresponsiveness; airway inflammation; asthmatic; base; beta catenin; chemokine; extracellular; immunoregulation; improved outcome; in vivo; macrophage; migration; mouse model; novel; overexpression; response; selective expression; therapeutic development; tool; transcription factor; versican; wound healing

Project Summary It is now recognized that the extracellular matrix (ECM) provides a bioactive environment in lungs (and other organs) that orchestrates cellular function for tissue morphogenesis, homeostasis, wound healing, and disease. Our published work and preliminary data using airway biopsies from subjects with and without asthma, mouse models, and in vitro cell cultures show that the ECM molecule, versican (gene = Vcan), accumulates in the lower airways and is strongly correlated with inflammation and airway hyperresponsiveness (AHR). We have found that versican is synthesized in response to allergens from at least two sources: fibroblasts and macrophages and that versican has a dramatic effect on leukocyte phenotype, adhesion and migration. To better understand the mechanisms whereby versican modulates the immune response to allergens, we propose to use two novel strains of mice with conditional deficiency in versican. These include Vcan-/- mice, which have a global deletion of versican when treated with tamoxifen; and LysM/Vcan-/- mice which lack versican in myeloid cells. The Vcan-/- and LysM/Vcan-/- mice have strikingly different pulmonary inflammatory responses, as summarized by three major findings: 1) stromal cells (e.g., fibroblasts) synthesize a form of versican that promotes the development of a pro-inflammatory ECM; 2) macrophage-derived versican restrains airway inflammation, which our preliminary data suggest is conferred by the selective expression of the V3 isoform of versican by macrophages; and 3) signaling pathways regulating versican expression in fibroblasts and macrophages differ, with the WNT/β-catenin/T-cell factor (TCF) pathway regulating Vcan expression in fibroblasts and, as we recently defined, the TLR/Trif/Type I interferon (IFN) pathway in macrophages. Our preliminary and published observations underscore the importance of understanding the immunomodulatory properties of versican and are the basis for our central hypothesis that allergens such as HDM increase the production of fibroblast-derived versican forming a pro-inflammatory ECM that promotes airway inflammation and AHR leading to chronic asthmatic conditions. Conversely, macrophage- derived versican forms an anti-inflammatory ECM. This hypothesis will be tested through completion of our two aims: Aim 1. Identify the role of fibroblast- vs. macrophage-derived versican in airway inflammation and AHR caused by HDM allergen and the mechanisms regulating Vcan expression in these cells. Aim 2. Define the mechanisms by which the structural components of versican promote or restrain airway inflammation through the formation of a pro- or anti-inflammatory ECM. Understanding the contextual settings in which versican provides fine control of airway inflammation and AHR is critical for the development of therapeutic strategies for improving outcomes in patients with asthma.

项目概要 现在人们认识到细胞外基质 (ECM) 在肺（和其他器官）中提供了生物活性环境。 器官）协调细胞功能以实现组织形态发生、体内平衡、伤口愈合和 疾病。我们发表的工作和初步数据使用了患有和不患有这种疾病的受试者的气道活检 哮喘、小鼠模型和体外细胞培养表明 ECM 分子 versican（基因 = Vcan） 积聚在下呼吸道，与炎症和气道高反应性密切相关 （AHR）。我们发现多功能蛋白聚糖是针对至少两个来源的过敏原​​而合成的： 成纤维细胞和巨噬细胞，多功能蛋白聚糖对白细胞表型、粘附和 迁移。为了更好地了解多功能蛋白聚糖调节免疫反应的机制 过敏原，我们建议使用两种具有条件性多功能蛋白聚糖缺陷的新型小鼠品系。这些包括 Vcan-/- 小鼠，在接受他莫昔芬治疗时，其 versican 整体缺失；和 LysM/Vcan-/- 小鼠 骨髓细胞中缺乏多功能蛋白聚糖。 Vcan-/- 和 LysM/Vcan-/- 小鼠的肺功能显着不同 炎症反应，总结为三个主要发现：1）基质细胞（例如成纤维细胞）合成 一种多功能蛋白聚糖，可促进促炎 ECM 的发展； 2) 巨噬细胞来源 Versacan 抑制气道炎症，我们的初步数据表明，这是由选择性 巨噬细胞表达多功能蛋白聚糖的V3亚型； 3) 调节多功能蛋白聚糖的信号通路 成纤维细胞和巨噬细胞中的表达存在差异，WNT/β-连环蛋白/T 细胞因子 (TCF) 途径 调节成纤维细胞中的 Vcan 表达，以及我们最近定义的 TLR/Trif/I 型干扰素 (IFN) 巨噬细胞中的通路。我们的初步和已发表的观察结果强调了 了解多功能蛋白聚糖的免疫调节特性，这是我们中心假设的基础： HDM 等过敏原会增加成纤维细胞衍生的多功能蛋白聚糖的产生，形成促炎 ECM 促进气道炎症和 AHR，导致慢性哮喘病。相反，巨噬细胞- 衍生的多功能蛋白聚糖形成抗炎 ECM。这个假设将通过完成我们的两个实验来检验 目标：目标 1. 确定成纤维细胞与巨噬细胞来源的多功能蛋白聚糖在气道炎症和 AHR 中的作用 由 HDM 过敏原和调节这些细胞中 Vcan 表达的机制引起。目标 2. 定义 多功能蛋白聚糖的结构成分通过以下机制促进或抑制气道炎症 促炎或抗炎 ECM 的形成。了解 versican 所处的情境设置 提供对气道炎症的精细控制，AHR 对于治疗策略的制定至关重要 改善哮喘患者的预后。