Corneal Metalloproteinase-Matrix Interactions

角膜金属蛋白酶-基质相互作用

• 批准号: 7647778
• 负责人: Mark I Rosenblatt
• 金额: $ 2.48万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7647778
• 关键词: 3-Dimensional; Biological Assay; Blindness; Cell Line; Cells; Cicatrix; Cleaved cell; Collagen Type IV; Condition; Cornea; Corneal Injury; Data; Deposition; Disease; End Point; Endopeptidases; Enzymes; Epithelium; Extracellular Matrix; Gel; Gene Activation; Genes; Goals; Green Fluorescent Proteins; Healed; Immunohistochemistry; In Vitro; Incubated; Injury; Investigation; Knock-in Mouse; Knockout Mice; Laboratories; Laser injury; Lasers; Localized; Mass Spectrum Analysis; Matrilysin; Matrix Metalloproteinases; Measures; Migration Assay; Mus; N-terminal; Organ; Peptide Hydrolases; Polymerase Chain Reaction; Preparation; Proteins; Regulation; Role; Site; Substrate Specificity; System; Techniques; Tenascin; Time; Transcriptional Activation; Transgenic Animals; Transgenic Mice; Translating; Western Blotting; Wild Type Mouse; Wound Healing; base; corneal epithelium; decorin; enzyme activity; healing; in vitro Assay; insight; migration; prevent; promatrilysin; promoter; protein expression; research study; two-dimensional; wound

DESCRIPTION (provided by applicant): Loss of corneal clarity is a common endpoint of many blinding conditions. Corneal opacification is a major cause of blindness worldwide. Our laboratory is studying the mechanisms associated with corneal wound repair that regulate coneal clarity. Specifically, we have identified an endogenous matrix metalloproteinase within the cornea which helps prevent corneal opcification after an excimer laser wound. Matrilysin (MMP-7) deficient mice have exuberant scarring following laser injuries which are innocuous to their wild-type counterparts. In this application we will investigate the interaction of MMP-7 with extracelluar matrix (ECM) components localized to native and wounded corneas. Specific Aim A will use a combination of immunohistochemistry, western blotting, zymography, and transgenic animals to localize the sites and quantify the amount of MMP-7 expressed in wounded and unwounded corneas. Specific Aim B extends these studies to determine the matrix substrate specificities of corneal epithelium- and keratocyte-derived MMP-7 in vitro. In this aim we will examine corneal celI-ECM interactions by assaying the migration of epithelium on 2-dimensional matrix gels, and the proliferation of keratocytes in 3-dimensional matrix gels. Specific Aim C will translate the in vitro results to MMP-7 knockout mice. The expression of xtracellular matrix components in wild-type and matrilysin knockout mice after excimer laser wounding will be evaluated by immunohistochemistry, western blotting and real-time PCR. The second portion of Aim C will carefully examine the MMP-7 dependent breakdown products of corneal extracellular matrix in these mice. Lastly, using MMP-7/ECM double knockout mice, the requirement of specific extracellular matrix components for reduced corneal clarity will be investigated. Our studies of protease-matrix interactions in wounded corneas will further our understanding of the mechanisms for maintaining corneal clarity. Results of these studies in the cornea may ultimately be helpful in understanding healing in other organs as well.

描述（由申请人提供）：角膜清晰度的丧失是许多致盲条件的常见终点。角膜混浊是全世界失明的主要原因。我们的实验室正在研究与角膜伤口修复相关的机制，以调节角膜清晰度。具体来说，我们已经在角膜内发现了一种内源性基质金属蛋白酶，它有助于防止准分子激光伤后角膜混浊。母体溶解素（MMP-7）缺陷小鼠在激光损伤后会产生大量疤痕，而这些疤痕对野生型小鼠是无害的。