Expression of Heparan Sulfate 6-O-Endosulfatases in Idiopathic Pulmonary Fibrosis

硫酸乙酰肝素6-O-内切硫酸酯酶在特发性肺纤维化中的表达

基本信息

批准号：
7837606
负责人：
XINPING YUE
金额：
$ 7.45万
依托单位：
TULANE UNIVERSITY OF LOUISIANA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-11 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7837606
关键词：
A549 Address Adenocarcinoma Cell Alveolar Animal Model Basement membrane Cell Adhesion Molecules Cell Culture Techniques Cell surface Cells Data Development Disaccharides Disease Epithelial Cells Epithelium Etiology Event Fibroblasts Gene Chips Gene Expression Genes Glycosaminoglycans Growth Factor Hamman-Rich syndrome Heparan Sulfate Proteoglycan Heparitin Sulfate Human Hyperplasia Immunohistochemistry Impairment In Situ Hybridization Inorganic Sulfates Interstitial Lung Diseases Knowledge Laboratories Lead Lung Maps Measures Messenger RNA Modeling Molecular Molecular Profiling Mus Organ Pathogenesis Patients Pattern Positioning Attribute Proteins Proteoglycan Research Reverse Transcriptase Polymerase Chain Reaction Role Sampling Sentinel Serum Severities Severity of illness Signaling Molecule Site Staging Structure Structure of parenchyma of lung Sulfatases Testing Time Tissues Transforming Growth Factors United States Unspecified or Sulfate Ion Sulfates Western Blotting Work abstracting alveolar type II cell cell type chemokine cytokine effective therapy laser capture microdissection morphogens new therapeutic target novel novel therapeutic intervention overexpression public health relevance pulmonary function spatial relationship sulfation

项目摘要

DESCRIPTION (provided by applicant): Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease of unknown etiology, characterized by scaring in the lung and impaired pulmonary function. The pathogenetic mechanisms leading to IPF are poorly understood, and there are currently no effective therapies. Characterization of the molecular and cellular changes occurring in the lung that can be correlated, not only with disease severity, but also with tissue structure and specific cell types is invaluable for the identification of new therapeutic targets. Data from both patients and animal models support a pivotal role for transforming growth factor (TGF)-21 in the development and progression of IPF. Our laboratory has made the sentinel observation that TGF-21 induces heparan sulfate (HS) 6-O-endosulfatase 1 and 2 (Sulf1 and Sulf2), which are novel cell surface sulfatases that remove the 6-O-sulfates from specific HS intra-chain sites. HS is the glycosaminoglycan moiety of the HS proteoglycans (HSPGs), which are major components of the cell surface, basement membrane and the extracelluar matrix. HSPGs interact with and modulate the activity of a myriad of signaling molecules through their HS chains, and it is now well established that HS-protein interactions critically depend on the amount and the positions of the O-sulfate groups, in particular, the 6-O-sulfates. Considering the importance of HS as both a structural and a signaling molecule, and the fact that TGF-21 induces Sulf1 and Sulf2, it is clear that detailed characterizations of these sulfatases in human IPF are important to elucidate their roles in the pathogenesis of this devastating illness. Using human IPF samples from the lung tissue research consortium (LTRC), we propose to test the following hypothesis in this study: Heparan sulfate 6-O-endosulfatases, Sulf1 and Sulf2, are involved in the pathogenesis of idiopathic pulmonary fibrosis. The specific aims are: 1) Examine Sulf1 and Sulf2 expression in control and IPF lungs using real-time quantitative RT-PCR and Western blotting; 2) Examine Sulf1 and Sulf2 localization in control and IPF lungs using in situ hybridization and immunohistochemistry; 3) Examine the Glyco-gene expression profiles of Sulf1 and Sulf2 expressing cells as well as cells from fibroblastic foci and type II hyperplasia using laser-capture microdissection and Glyco-gene chip array. To our knowledge, this is the first study to address the involvement of Sulf1 and Sulf2 in the pathogenesis of IPF. Our findings could lead to the development of new therapeutic interventions for IPF as well as fibrogenic diseases in other organs. PUBLIC HEALTH RELEVANCE: There are currently over 200,000 patients in the United States and five million people worldwide suffering from idiopathic pulmonary fibrosis (IPF). Examination of Sulf1 and Sulf2 expression in human IPF samples will reveal whether and how Sulf1 and Sulf2 are involved in the pathogenesis of this devastating illness. Findings from this study will help us better understand the molecular mechanisms leading to IPF, and identify new therapeutic targets in treating this disease. (End of Abstract)

描述（由申请人提供）：特发性肺纤维化（IPF）是一种病因不明的进行性间质性肺疾病，其特征是肺部疤痕和肺功能受损。导致IPF的发病机制尚不清楚，目前尚无有效的治疗方法。肺部发生的分子和细胞变化的表征不仅与疾病的严重程度相关，而且与组织结构和特定细胞类型相关，对于识别新的治疗靶点具有无价的价值。来自患者和动物模型的数据支持转化生长因子 (TGF)-21 在 IPF 的发生和进展中发挥关键作用。我们实验室进行了前哨观察，TGF-21 诱导硫酸乙酰肝素 (HS) 6-O-内切硫酸酯酶 1 和 2（Sulf1 和 Sulf2），它们是新型细胞表面硫酸酯酶，可去除特定 HS 内的 6-O-硫酸酯。连锁网站。 HS 是 HS 蛋白聚糖 (HSPG) 的糖胺聚糖部分，是细胞表面、基底膜和细胞外基质的主要成分。 HSPG 通过其 HS 链与无数信号分子相互作用并调节其活性，现在已经确定 HS-蛋白质相互作用关键取决于 O-硫酸盐基团的数量和位置，特别是 6- O-硫酸盐。考虑到 HS 作为结构分子和信号分子的重要性，以及 TGF-21 诱导 Sulf1 和 Sulf2 的事实，很明显，人类 IPF 中这些硫酸酯酶的详细特征对于阐明它们在这种毁灭性的发病机制中的作用非常重要。疾病。使用来自肺组织研究联盟 (LTRC) 的人类 IPF 样本，我们建议在本研究中检验以下假设：硫酸乙酰肝素 6-O-内切硫酸酯酶 Sulf1 和 Sulf2 参与特发性肺纤维化的发病机制。具体目标是： 1) 使用实时定量 RT-PCR 和蛋白质印迹检查对照和 IPF 肺中 Sulf1 和 Sulf2 的表达； 2) 使用原位杂交和免疫组织化学检查 Sulf1 和 Sulf2 在对照和 IPF 肺部的定位； 3) 使用激光捕获显微切割和糖基因芯片阵列检查 Sulf1 和 Sulf2 表达细胞以及来自成纤维细胞灶和 II 型增生的细胞的糖基因表达谱。据我们所知，这是第一项探讨 Sulf1 和 Sulf2 参与 IPF 发病机制的研究。我们的研究结果可能会导致针对 IPF 以及其他器官纤维形成疾病的新治疗干预措施的开发。公共卫生相关性：目前美国有超过 200,000 名患者，全球有 500 万人患有特发性肺纤维化 (IPF)。对人类 IPF 样本中 Sulf1 和 Sulf2 表达的检查将揭示 Sulf1 和 Sulf2 是否以及如何参与这种毁灭性疾病的发病机制。这项研究的结果将帮助我们更好地了解导致IPF的分子机制，并确定治疗这种疾病的新治疗靶点。（摘要完）