Sulfs and Injury Repair in Mucosal Epithelia

粘膜上皮中的硫和损伤修复

• 批准号: 7556200
• 负责人: STEVEN D ROSEN
• 金额: $ 17.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-25 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7556200
• 关键词: 3-Dimensional; A549; Acute Lung Injury; Adherent Culture; Agonist; Alveolar; Alveolar Cell; Alveolar wall; Antibodies; Bacterial Adhesins; Binding; Catabolism; Cell Line; Cell Proliferation; Cell surface; Cells; Collaborations; Complement; Cornea; Cultured Cells; Cyst; Development; Distal; Enzyme Activation; Enzymes; Epithelial; Epithelial Cells; Epithelium; Event; Extracellular Matrix; Eye; GAG Gene; Gene Targeting; Glucosamine; Growth Factor; Heparan Sulfate Proteoglycan; Human; Image; In Vitro; Infection; Inflammatory; Injury; Knockout Mice; Ligand Binding; Ligands; Lung; Malignant Neoplasms; Malignant neoplasm of pancreas; Matrix Metalloproteinases; Metalloproteases; Methods; Modeling; Modification; Molecular; Mus; Muscle Development; Organ Culture Techniques; Pathway interactions; Pattern; Process; Pseudomonas Infections; RNA Interference; Reagent; Regulation; Resources; Respiratory System; Role; Signal Transduction; Signaling Molecule; Site; Sulfatases; Testing; Tissues; Up-Regulation; Vascular Endothelial Growth Factors; Work; Wound Healing; alveolar epithelium; cell growth; cell motility; chemokine; corneal epithelium; corneal repair; extracellular; functional restoration; in vivo; in vivo Model; inhibitor/antagonist; injured; injury and repair; insight; macromolecule; microbial; migration; monolayer; morphogens; novel; pathogen; receptor; repaired; response; small molecule; sulfation; tool; wound

Project 3: Sulfs and Wound Repair in Mucosal Epithelium Mucosal epithelia covering the respiratory tract and cornea are vulnerable sites for the entry of pathogens and noxious substances. Upon injury, these epithelia must be rapidly repaired to restore function and to limit infection. Our work will focus on two novel sulfatases, known as Sulf-1 and Sulf-2, which we have cloned and characterized in human and mouse. The Sulfs are extracellular endosulfatases, which act on heparan sulfate proteoglycans (HSPGs). HSPGs influence a huge variety of processes by binding to a multiplicity of signaling molecules such as chemokines, growth factors, and morphogens (e.g., Wnts). The Sulfs "edit" the sulfation status of HSPGs by selectively removing the 6-O-S from glucosamine within HS chains. In so doing, the Sulfs modulate HSPG-ligand interactions and regulate signaling events and infection. The Sulfs promote Wnt signaling in development and cancer. Since Wnt signaling is also involved in epithelial wound responses, we have investigated the role of the Sulfs in the repair of mucosal epithelia. Our studies reveal that Sulf is expressed in lung type II alveolar epithelial cells (AT-II), which are critical in the the reepithelialization of injured alveolar walls. Moreover, Sulf regulates Wnt signaling and cell growth of an alveolar epithelial cell line. We will investigate the involvement of Sulfs in the repair of scratched monolayers of cultured alveolar cells, as a model of acute lung injury. Our second focus will be wound repair of corneal epithelium. Here, we found that Sulf is dramatically upregulated upon scratching of mouse cornea. In a cell culture model of human corneal epithelial cells, Sulf is required for repair of scratched monolayers. Our proposed studies of wound repair of alveolar epithelium and corneal epithelium will take advantage of established and novel methods for inhibiting the activity and expression of the Sulfs, including RNAi, antibodies, a small molecule inhibitor of the Sulfs, and Sulf null mice (single and double). We hypothesize that the Sulfs are pivotal in epithelial repair processes, by promoting Wnt-dependent cell growth and cell migration. By understanding the roles of Sulfs in the repair of distal airways and cornea, new methods for enhancing wound repair of these highly vulnerable mucosal epithelia may emerge. Projects 1, 2, and 4 will investigate related aspects of the Sulfs. We will extensively colloborate with them and Cores B and C.

项目 3：粘膜上皮中的硫化物和伤口修复 覆盖呼吸道和角膜的粘膜上皮是病原体进入的脆弱部位 和有毒物质。受伤后，这些上皮必须迅速修复以恢复功能并限制 感染。我们的工作将集中于两种新型硫酸酯酶，即 Sulf-1 和 Sulf-2，我们已经克隆并开发了这两种酶。 以人类和小鼠为特征。 Sulfs 是细胞外硫酸内酯酶，作用于乙酰肝素 硫酸盐蛋白聚糖（HSPG）。 HSPG 通过与多种物质结合来影响多种过程。 信号分子，例如趋化因子、生长因子和形态发生素（例如 Wnts）。苏尔夫“编辑”了 通过选择性地去除 HS 链内葡萄糖胺中的 6-O-S 来检测 HSPG 的硫酸化状态。在这样 Sulfs 调节 HSPG-配体相互作用并调节信号事件和感染。苏尔夫人 促进发育和癌症中的 Wnt 信号传导。由于 Wnt 信号传导也参与上皮伤口 响应，我们研究了 Sulfs 在粘膜上皮修复中的作用。我们的研究表明 Sulf 在肺 II 型肺泡上皮细胞 (AT-II) 中表达，这对于上皮再形成至关重要 受伤的肺泡壁。此外，Sulf 还调节 Wnt 信号传导和细胞生长 肺泡上皮细胞系。我们将研究硫化物在修复划痕单层细胞中的作用 培养的肺泡细胞，作为急性肺损伤的模型。我们的第二个重点是角膜伤口修复 上皮。在这里，我们发现当刮擦小鼠角膜时，硫含量显着上调。在一个牢房里 在人角膜上皮细胞培养模型中，修复划伤的单层细胞需要硫。我们的 拟议的肺泡上皮和角膜上皮伤口修复研究将利用 已建立的抑制 Sulfs 活性和表达的新方法，包括 RNAi， 抗体、Sulf 的小分子抑制剂和 Sulf 无效小鼠（单小鼠和双小鼠）。我们假设 Sulfs 通过促进 Wnt 依赖性细胞生长和细胞增殖，在上皮修复过程中发挥关键作用 迁移。通过了解 Sulfs 在修复远端气道和角膜中的作用，新方法 可能会出现增强这些高度脆弱的粘膜上皮的伤口修复。项目 1、2 和 4 将 调查Sulfs的相关方面。我们将与他们以及核心 B 和 C 进行广泛的合作。