Mechanisms and Consequences of Hyaluronan Production in Asthma

哮喘中透明质酸产生的机制和后果

• 批准号: 9232189
• 负责人: MARK A ARONICA
• 金额: $ 32.79万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9232189
• 关键词: Activities of Daily Living; Adhesives; Air; Apical; Asthma; Award; Basement membrane; Biological Assay; Cell Culture Techniques; Cells; Chronic; Clinical; Coculture Techniques; Coupled; Coupling; Data; Development; Epithelial; Epithelial Cells; Epithelium; Extracellular Matrix; Functional disorder; Future; Genes; HAS2 gene; Human; Hyaluronan; Inflammation; Inflammatory Response; Interferons; Interleukin-1; Interleukin-13; Knockout Mice; Lead; Leukocytes; Liquid substance; Lung; Mediating; Mediator of activation protein; Metabolism; Methods; Mus; Muscle Cells; NOS1 gene; NOS2A gene; Nitric Oxide; Nitric Oxide Pathway; Nitric Oxide Synthase; Pathologic; Pathway interactions; Phosphotransferases; Play; Poly I-C; Production; Proteins; Regulation; Resources; Role; Sampling; Signal Transduction; Slice; Smooth Muscle; Soluble Guanylate Cyclase; Sulfhydryl Compounds; System; TNF gene; Testing; Therapeutic Intervention; Time; Viral; Work; airway epithelium; airway hyperresponsiveness; airway inflammation; airway remodeling; asthmatic; asthmatic airway; cytokine; endoplasmic reticulum stress; experimental study; in vivo; inhibitor/antagonist; innovation; member; mimetics; mouse model; novel; programs; respiratory smooth muscle; skills

ABSTRACT - PROJECT 2 During the previous award cycle, we identified mechanisms by which primary airway epithelial cells (AECs) synthesize heavy chain (HC) modified hyaluronan (HA) apical rafts, and airway smooth muscle cells (ASMCs) synthesize an extracellular matrix of HA cables, which are leukocyte-adhesive or what we have also termed as pathologic-HA. Our studies in the Program revealed the importance of this pathologic-HA matrix in the genesis of inflammation, airway remodeling and hyperresponsiveness of asthma. Within the airway, there is constant interaction between the closely coupled ASMCs and overlying epithelium, and increasing data indicate a primary role for epithelium and smooth muscle in the origins of inflammation through production of early mediators. In this continuation of Project 2, we propose to investigate if airway epithelial production of NO and cytokines promotes the accumulation of a pathologic-HA matrix by the smooth muscle in asthma. In preliminary data, we show that the airway epithelium responds to prototypic Th2 cytokines or cytokine mixes with the production of HA and induction of iNOS. Utilizing our novel co-culture system, we show that the ASMCs can respond to epithelial derived NO with the production of leukocyte-adhesive HA cables. We also show that the major contributor to HA production is via the hyaluronan synthase 2 (HAS2). Collaborative studies of NO regulation and signaling mechanisms, which are investigated by other members of this Program, support our experiments to define mechanisms of pathological HA formation, including NO metabolism (Project 1), specific protein thiol nitrosylation (Project 3), and NO activation pathways (Project 4). Thus, in this continuation, we propose to test the hypothesis that NO and HCs produced by the asthmatic airway epithelium promote formation of a pathologic, leukocyte-adhesive HC-HA matrix by ASMCs, through mechanisms that appear to be sGC-independent NO recognition pathways. To test this hypothesis, we plan to identify pathways leading to airway epithelial cell production of HA [Aim 1]. We also plan to identify pathways of ASMC production of a leukocyte-adhesive HA matrix [Aim 2]. Finally, we will determine the contribution of NO and HA on the development of inflammation, AHR, and remodeling in vivo [Aim 3]. Project 2 depends on the collaborative interactions and complementary skills, expertise, and resources of the Program to test hypotheses. Program Cores support novel murine models, technically challenging assays, and shared clinical samples in order to facilitate translational and mechanistic experiments in Project 2 not otherwise possible.

摘要 - 项目2 在上一个奖励周期中，我们确定了主要气道上皮细胞（AEC）的机制 合成重链（HC）修饰的透明质酸（HA）顶筏和气道平滑肌细胞（ASMC） 合成HA电缆的细胞外基质，该基质是白细胞粘附的，或者我们也称为 病理性ha。我们在该计划中的研究揭示了该病理学基质在创世纪的重要性 炎症，气道重塑和哮喘的反应性过高。在气道中，有恒定 紧密耦合的ASMC与上皮上皮之间的相互作用，并增加数据表明A 上皮和平滑肌的主要作用在炎症的起源中通过早期产生 调解人。在项目2的延续中，我们建议调查气道上皮生产不和 细胞因子促进哮喘中平滑肌的病理-HA基质的积累。在 初步数据，我们表明气道上皮对原型TH2细胞因子或细胞因子混合物做出反应 随着HA的产生和iNOS诱导。利用我们的新型共培养系统，我们证明了 ASMC可以通过产生白细胞粘附HA电缆的上皮响应。我们也是 表明HA生产的主要贡献者是通过透明质酸合酶2（HAS2）。协作 该计划的其他成员研究的没有调节和信号传导机制的研究， 支持我们的实验来定义病理HA形成的机制，包括没有新陈代谢（项目 1），特异性蛋白硫醇亚硝基化（项目3），没有激活途径（项目4）。因此，在此 继续，我们建议测试哮喘气道产生的NO和HCS的假设 上皮促进通过ASMC的病理，白细胞粘附的HC-HA基质形成 似乎是独立于SGC的机制，没有识别途径。为了检验这一假设，我们 计划识别导致气道上皮细胞产生HA的途径[AIM 1]。我们还计划确定 ASMC产生白细胞粘合剂HA矩阵的途径[AIM 2]。最后，我们将确定 NO和HA对体内炎症，AHR和重塑的发展的贡献[AIM 3]。项目2 取决于该计划的协作互动和互补技能，专业知识和资源 检验假设。计划核心支持新颖的鼠模型，技术挑战性的测定和共享 临床样本以促进项目2中的翻译和机械实验 可能的。