The Role of Extracelluar Enzymes in Regulating Corneal Repair

细胞外酶在调节角膜修复中的作用

• 批准号: 8048007
• 负责人: Matilda F Chan
• 金额: $ 22.07万
• 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: 2008-04-01 至 2013-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8048007
• 关键词: Ablation; Actins; Address; Affect; Angiogenesis Inhibitors; Angiogenic Factor; Angiostatins; Behavior; Binding; Biology; Blindness; Bone Marrow Transplantation; Cells; Clinical; Confocal Microscopy; Cornea; Corneal Injury; Corneal Neovascularization; Corneal Stroma; Development; Disease; Enzyme Inhibition; Enzymes; Epithelial; Epithelial Cell Proliferation; Epithelium; Equilibrium; Event; Extracellular Matrix; Fibroblast Growth Factor; Fibronectins; Fibrosis; Gelatinase B; Generations; Growth Factor; Image; Imaging Techniques; In Vitro; Inflammatory; Inflammatory Response; Injury; Interstitial Collagenase; Keratoplasty; Knockout Mice; Label; Laser Surgery; Lead; Maintenance; Matrix Metalloproteinases; Mediating; Mentors; Mus; Mutant Strains Mice; Myeloid Cells; Pathologic; Pathway interactions; Phenotype; Play; Process; Recruitment Activity; Regulation; Relative (related person); Research Personnel; Role; Scientist; Site; Stress Fibers; Sulfatases; Time; Trauma; Ulcer; Vision; angiogenesis; antiangiogenesis therapy; base; career development; chemokine; corneal epithelium; corneal repair; cytokine; design; extracellular; in vitro Assay; in vivo; insight; macrophage; matrix metalloproteinase 12; novel; prevent; programs; repaired; response; response to injury; therapeutic target

DESCRIPTION (provided by applicant): Corneal opacification affects millions of people and is the second leading cause of blindness in the world. A clear need exists for the development of non-surgical strategies to prevent and treat corneal fibrosis and the subsequent development of corneal opacification. Previous and preliminary studies show that extracellular enymes including the 6-O-sulfatase Sulf-1 and matrix metalloproteinases (MMPs) including MMP-9 and MMP-12 are expressed in the cornea only upon injury, implicating their possible roles in corneal repair. These enzymes are produced by epithelial, stromal and inflammatory cells, and their roles in corneal repair c have not been well-studied. In the epithelium, MMP-9 appears to decrease re-epithelialization through its effects on the TGF pathway, while in the stroma MMP-12 has been shown to be protective against fibrosis via TGF-p in various other disease processes. In addition, MMP-12's ability to produce angiostatin more efficiently than other MMPs suggests that it might play a role in preventing corneal angiogenesis. Sulf-1 has been found to modulate the binding of growth factors and chemokines in vitro. Stimulation of various growth factors, particularly TGF-p2 and FGF, are critical to the corneal fibrotic phenotype and corneal neovascularization. Taken together, these extracellular enzymes may regulate various aspects of the repair process, both positively and negatively. In this application, we propose to investigate the roles of the extracellular enzymes MMP-9, MMP-12, and Sulf-1 in the epithelial, stromal, and inflammatory corneal response to injury. Specific Aim 1 will determine the effects of MMP-9 or Sulf-1 activity on the epithelial cell proliferation response to injury. Specific Aim 2 will use MMP-12 knockout mice to examine the role of MMP- 12 in fibrosis and angiogenesis during the corneal stromal response to injury. Specific Aim 3 will examine the ability of these extracellular enzymes to recruit inflammatory cells to the cornea and will use a novel imaging technique based on spinning disk confocal microscopy to study inflammatory cell dynamics in the cornea in vivo and in real-time. Understanding the roles of extracellular enzymes in wounded corneas should further our understanding of the mechanisms important to the maintenance of corneal clarity and provide basic insights that will, in the long term, allow for the development of better treatment of corneal opacification.

描述（由申请人提供）：角膜混浊影响数百万人，是世界上第二大失明原因。显然需要开发非手术策略来预防和治疗角膜纤维化以及随后出现的角膜混浊。先前的初步研究表明，细胞外酶（包括 6-O-硫酸酯酶 Sulf-1）和基质金属蛋白酶（MMP）（包括 MMP-9 和 MMP-12）仅在角膜损伤时表达，这表明它们在角膜修复中可能发挥作用。这些酶由上皮细胞、基质细胞和炎症细胞产生，它们在角膜修复中的作用尚未得到充分研究。在上皮细胞中，MMP-9 似乎通过其对 TGF 通路的影响来减少再上皮化，而在基质中，MMP-12 已被证明可以在各种其他疾病过程中通过 TGF-β 预防纤维化。此外，MMP-12 比其他 MMP 更有效地产生血管抑制素的能力表明它可能在预防角膜血管生成中发挥作用。已发现 Sulf-1 可在体外调节生长因子和趋化因子的结合。各种生长因子的刺激，特别是 TGF-β2 和 FGF，对于角膜纤维化表型和角膜新生血管形成至关重要。总的来说，这些胞外酶可以积极和消极地调节修复过程的各个方面。在此应用中，我们打算研究细胞外酶 MMP-9、MMP-12 和 Sulf-1 在上皮、基质和炎症角膜损伤反应中的作用。具体目标 1 将确定 MMP-9 或 Sulf-1 活性对上皮细胞增殖损伤反应的影响。具体目标 2 将使用 MMP-12 敲除小鼠来检查 MMP-12 在角膜基质损伤反应过程中纤维化和血管生成中的作用。具体目标 3 将检查这些细胞外酶将炎症细胞募集到角膜的能力，并将使用基于转盘共聚焦显微镜的新型成像技术来体内实时研究角膜中的炎症细胞动力学。了解细胞外酶在受伤角膜中的作用应该可以进一步加深我们对维持角膜透明度的重要机制的理解，并提供基本的见解，从长远来看，可以更好地治疗角膜混浊。