Role of Heparan Sulfate NDST-1 in Allergic Inflammation and Airway Remodeling

硫酸乙酰肝素 NDST-1 在过敏性炎症和气道重塑中的作用

• 批准号: 7150796
• 负责人: P. SRIRAMARAO
• 金额: $ 39.59万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7150796
• 关键词: T lymphocyte; angiogenesis; asthma; cell migration; cooperative study; enzyme activity; enzyme linked immunosorbent assay; genetically modified animals; heparan sulfate; histology; hypersensitivity; inflammation; laboratory mouse; leukocytes; macrophage; respiratory disorder; respiratory function; stem cells; sulfotransferase

Allergic asthma is a chronic inflammatory disorder of the airways. There is good evidence that many of the features of asthma, including airway remodeling (AR) are largely due to inappropriate activation of T cells and consequent imbalance of Th1/Th2 cytokines and the recruitment if inflammatory cells including macrophages to the airway. However, the role of heparan sulfates (HS) in the modulation of airway allergic responses and airway remodeling are poorly understood. Heparin and HS interact with various components of the inflammatory cascade in vitro, such as P- and L-selectins, inflammatory mediators, proteoglycans, growth factors cytokines and chemokines to mediate leukocyte migration, endothelial proliferation and tissue remodeling. However the role of endothelial and leukocyte expressed HS in allergic inflammation and AR has never been determined. To study the significance of HS in AR, glucosaminyl N-deacetylase/N-sulfotransferase- 1 (NDST1) was knocked out in endothelial cells (EC) and leukocytes using the Cre-loxP system. The NDST-1-deficient mice (NDST1f/fTie2Cre+) showed impaired inflammatory responses in multiple types of assays in vitro and in vivo. Based on our preliminary observations that lack of NDST-1 in allergenchallenged mice leads to altered leukocyte trafficking, impaired macrophage but not eosinophil recruitment in the airways, reduced expression of the Th2 cytokine IL-5, TGF-beta1 and AHR, along with diminished EC proliferation, we postulate that the targeted deficiency of NDST-1 will result in abrogation of the inflammatory conditions associated with allergen-induced inflammation and AR. To examine this, two specific aims are proposed. In the first specific aim we will assess the role of HS biosynthetic enzyme NDST-1 in the expression of Th2 cytokines, generation and recruitment of macrophages and macrophage progenitor cells to the airways and trafficking of mononuclear cells within the pulmonary vascular bed in the remodeled airways of allergen mice. In this aim the ability of bone marrow derived CD34+ progenitors cells to interact with murine lung EC in the context of HS expression will be examined in NDST1f/fTie2Cre+ mutant and NDST1f/fTie2Cre- wild type mice. In the second specific aim we will delineate the role of HS/NDST-1 in allergen-mediated AR and pulmonary angiogenesis associated with repetitive allergen challenge. Specifically, we will examine the role of HS/NDST-1 on the release of TGF-beta1, MMP-9, FGF-2 and VEGF by macrophages and other inflammatory cells and their effect on the cellular features of airway remodeling. We will also examine the role of NDST-1 and HS to modulate trafficking of inflammatory leukocytes with angiogenic blood vessels of the NDST-1 deficient and wild type mice exposed to repetitive allergen challgenge. Overall we will attempt to delineate the functional importance of EC vs. leukocyte expressed HS by NDST-1 in airway remodeling and airway allergic inflammation.

过敏性哮喘是一种慢性气道炎症性疾病。有充分的证据表明， 哮喘的特征，包括气道重塑（AR），很大程度上是由于T细胞的不适当激活 以及随后的Th 1/Th 2细胞因子的失衡和如果炎性细胞包括 巨噬细胞进入气道然而，硫酸乙酰肝素（HS）在调节气道过敏性哮喘中的作用尚不清楚。 反应和气道重塑知之甚少。肝素和HS与各种组分相互作用 体外炎症级联反应，如P-和L-选择素，炎症介质，蛋白聚糖， 生长因子细胞因子和趋化因子介导白细胞迁移、内皮细胞增殖和组织 重塑然而，内皮细胞和白细胞表达HS在变应性炎症和AR中的作用 从未被确定。为探讨HS在变应性鼻炎中的意义，采用葡萄糖氨基N-脱乙酰基酶/N-磺基转移酶- 1（NDST 1）在内皮细胞（EC）和白细胞中使用Cre-loxP 系统NDST-1缺陷小鼠（NDST 1f/fTie 2Cre+）在多个组织中显示出受损的炎症反应。 体外和体内试验的类型。根据我们的初步观察，NDST-1的缺乏在过敏原激发的 小鼠导致白细胞运输改变，巨噬细胞受损，但不是嗜酸性粒细胞招募， 气道中Th 2细胞因子IL-5、TGF-β 1和AHR的表达减少，沿着EC减少 因此，我们推测NDST-1的靶向缺陷将导致炎症反应的消除。 与过敏原诱导的炎症和AR相关的病症。为了检验这一点，有两个具体目标： 提出了在第一个具体目标中，我们将评估HS生物合成酶NDST-1在细胞内的作用。 Th 2细胞因子的表达，巨噬细胞和巨噬细胞祖细胞的产生和募集 在重塑的肺血管床中， 过敏原小鼠的气道。在这个目标中，骨髓来源的CD 34+祖细胞与人的骨髓细胞相互作用的能力被认为是一种免疫调节剂。 将在NDST 1f/fTie 2Cre+突变体和 NDST 1f/fTie 2Cre-野生型小鼠。在第二个具体目标中，我们将描述HS/NDST-1在以下方面的作用： 过敏原介导的AR和与反复过敏原激发相关的肺血管生成。 具体而言，我们将研究HS/NDST-1对巨噬细胞和其他炎症细胞释放TGF-β 1，MMP-9，FGF-2和VEGF的作用及其对气道重塑细胞特征的影响。我们 还将研究NDST-1和HS调节炎症性白细胞运输的作用， 暴露于重复变应原的NDST-1缺陷型和野生型小鼠的血管生成血管 你好总的来说，我们将试图描述EC与白细胞表达的HS的功能重要性。 NDST-1在气道重塑和气道过敏性炎症中的作用。