Hyaluronan in Pulmonary Fibrosis and Asthma

透明质酸在肺纤维化和哮喘中的作用

• 批准号: 8403438
• 负责人: Paul Wesley Noble
• 金额: $ 35.19万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8403438
• 关键词: Alveolar; Alveolar Macrophages; Asthma; Automobile Driving; Basement membrane; Behavior; Biological Models; CD44 Antigens; CD44 gene; Cell membrane; Cell surface; Cells; Chronic; Chronic lung disease; Data; Development; Disease Progression; Epithelial; Epithelial Cells; Extracellular Matrix; Fibroblasts; Fibrosis; Generations; Glycosaminoglycans; Goals; Hamman-Rich syndrome; Human; Hyaluronan; Inflammation; Inflammatory; Injury; Instruction; Integration Host Factors; Interleukin-1; Interleukin-13; Laboratories; Lung; Lung Inflammation; Lung diseases; Mesenchymal; Modeling; Molecular; Morbidity - disease rate; Mus; Myofibroblast; Pathologic; Patients; Phenotype; Pneumonia; Principal Investigator; Pulmonary Fibrosis; Regulation; Relative (related person); Role; Signal Transduction; Smooth Muscle Actin Staining Method; Structure of parenchyma of lung; System; TLR2 gene; TLR4 gene; Testing; Tissues; Transgenes; Transgenic Mice; Work; asthmatic patient; base; extracellular; human disease; hyaluronan synthase 1; in vivo; loss of function; lung injury; macromolecule; man; mortality; mouse model; novel; programs; promoter; receptor; response; senescence; surfactant deficiency

instrucfions): The molecular mechanisms that control tissue remodeling remain incompletely understood. This program has focused on elucidafing the role of extracellular matrix in regulafing lung inflammafion and fibrosis. A crifical unanswered question in the pathobiology of chronic lung diseases is what are the determinants of disease progression and irreversible loss of funcfion. Persistent lung inflammation and fibrosis is a cause of major morbidity and mortality whether it occurs in the ainways as in chronic asthma or in the interstifium as in idiopathic interstitial pneumonias such an idiopathic pulmonary fibrosis (IPF). We have generated a body of work that has identified the glycosaminoglycan hyaluronan (HA) and its cognate receptors as having important roles in regulating lung inflammation and fibrosis in the context of noninfectious lung injury. HA is an unusual macromolecule in that it is synthesized at the cell membrane and extruded into the extracellular milieu and becomes fragmented during inflammafion. Work from our laboratory over the last 17 years has identified distinct functions for HA in noninfectious lung injury depending on both the cellular context of its expression and the form in which it is presented to interacting cells. HA is modified in the context of lung inflammation and breakdown products accumulate that promote inflammation. We have also discovered that HA expressed on the cell surface of lung epithelial cells serves a protective function against noninfectious insults. In contrast, when myofibroblasts are directed to over-express hyaluronan synthase 2 (HAS2),.the result is a severe and progressive fibrodestructive lung disease after injury that causes significant mortality. One of the pathologic hallmarks of IPF is the destruction of basement membrane at the alveolar-endothelial interface and we began to investigate whether fibroblast invasion of matrix could be an important feature of unremitting fibrosis. We have made the exciting observation that fibroblasts isolated from transgenic mice that over-express HAS2 under the direction of the D-smooth muscle actin promoter have an invasive phenotype relative to transgene negative control fibroblasts. We have also developed the first model of targeted deletion of HAS2 in mesenchymal cells and found that the development of fibrosis following lung injury is impaired. In addifion, we have found a marked accumulation of HA in lung fissue using a novel chronic model of asthma in mice that causes ainway fibrosis. Collectively, this system offers unique approaches to dissect the mechanisms that control persistent fibrosis in two distinct model systems. Furthermore, we have also found that fibroblasts isolated from patients that have severe idiopathic pulmonary fibrosis and chronic asthma demonstrate an invasive phenotype. Based on these data from novel mouse models as well as human diseases we have generated the hypothesis that persistent inflammation and unremitting fibrosis is driven by both the accumulation of HA fragments and the development of an invasive fibroblast phenotype that requires hyaluronan synthase 2 and HA binding proteins such as CD44. We will test this hypothesis in the following Aims: Specific Aim 1. Determine the mechanisms by which over-expression of hyaluronan synthase 2 (HAS2) and CD44 regulate severe lung fibrosis and the development of an invasive fibroblast phenotype. Specific Aim 2. Determine the roles of hyaluronan synthase 2 (HAS2) and CD44 in regulafing the asthma phenotype and progressive ainway remodeling in a chronic model of mouse asthma using mice with targeted over-expression of HAS2 in myofibroblasts and deletion of HAS2 in lung epithelial and mesenchymal cells. Specific Aim 3. Determine the role of HAS2 and CD44 in regulating the invasive fibroblast phenotype in patients with IPF. RELEVANCE (See instructions): This project (Project 1) is an integral part of the global goal of this program to study the role of endogenous matrix components in driving unremitting lung inflammation and fitirosis by interacting with cognate receptors leading to the development of an invasive fibroblast phenotype. Project 1 will interface closely with Project 2 (Dr. Wright), as these concepts will be investigated in the context of surfactant deficiency. Project 1 will also interface closely with Project 3 (Dr. Kraft) to determine the roles of hyaluronan and its receptors in the pathobiology of asthma in mouse and man.

乐器）： 控制组织重塑的分子机制仍然不完全理解。这个程序 已重点是阐明细胞外基质在调节肺部炎症和纤维化中的作用。一个 在慢性肺疾病病理生物学中进行的未解决的问题是什么是决定因素 疾病的进展和不可逆转的功能丧失。持续的肺部炎症和纤维化是原因 主要发病率和死亡率是否发生在Ainways中，例如在慢性哮喘中或在 特发性间质性肺炎这样的特发性肺纤维化（IPF）。我们已经产生了一个 确定糖胺聚糖透明质酸（HA）及其同源受体的工作是 在非感染肺损伤的背景下调节肺部炎症和纤维化的重要作用。哈是 一种不寻常的大分子，因为它是在细胞膜上合成并挤出到细胞外的 环境，在炎症期间变得分散。过去17年来我们实验室的工作 确定了非感染性肺损伤中HA的不同功能，具体取决于其细胞的上下文 表达及其呈现给相互作用细胞的形式。 HA在肺的背景下进行了修改 炎症和分解产物会积累促进炎症。我们还发现 在肺上皮细胞的细胞表面表达的HA具有防护功能 侮辱。相比之下，当肌纤维细胞定向到过表达透明质酸合酶2（has2）时。 结果是损伤后严重而进行的纤维化肺部疾病，导致死亡率显着。 IPF的病理标志之一是肺泡 - 内皮的地下室膜的破坏 界面，我们开始研究成纤维细胞侵袭基质是否可能是 纤维化不舒服。我们已经做出了令人兴奋的观察结果，即从转基因小鼠中分离出的成纤维细胞 在D-Smooth肌肉肌动蛋白启动子的指示下，过表达的HAS2具有侵入性 表型相对于转基因阴性对照成纤维细胞。我们还开发了第一个模型 在间充质细胞中靶向删除HAS2，发现肺后纤维化的发展 伤害受损。另外，我们发现了使用新颖的HA在肺裂缝中明显的HA积累 小鼠哮喘的慢性模型，导致Ainway纤维化。总体而言，该系统提供了独特的 在两个不同模型系统中控制持续纤维化的机制的方法。 此外，我们还发现，从患有严重特发性的患者中分离出的成纤维细胞 肺纤维化和慢性哮喘表现出侵入性表型。 基于新的鼠标模型以及人类疾病的这些数据，我们已经生成了 假设持续的炎症和不舒服的纤维化均由两种积累驱动 HA碎片和需要透明质酸的侵入性成纤维细胞表型的发展 合酶2和HA结合蛋白，例如CD44。我们将在以下目的中检验这一假设： 具体目标1。确定透明质酸合酶2（HAS2）过表达的机制 和CD44调节严重的肺纤维化以及侵入性成纤维细胞表型的发展。 具体目标2。确定透明质酸合酶2（HAS2）和CD44在调节中的作用 哮喘表型和进行性AINWay在慢性小鼠哮喘中使用小鼠的慢性模型进行了重塑 在肌纤维细胞中has2的靶向过表达，并在肺上皮中删除HAS2和 间充质细胞。 特定目标3。确定HAS2和CD44在调节侵入性成纤维细胞表型中的作用 IPF患者。 相关性（请参阅说明）： 该项目（项目1）是研究内源性作用的全球目标的组成部分 通过与同源受体相互作用，在驱动肺部炎症和fitiorosis的基质成分中 导致侵入性成纤维细胞表型的发展。项目1将与项目2紧密接口 （赖特博士），因为这些概念将在表面活性剂缺乏症的背景下进行研究。项目1也将 与项目3（Kraft博士）紧密接触，以确定透明质酸及其受体在 小鼠和人类哮喘的病理生物学。