Identification of a new role of membrane‐type 1 matrix metalloproteinases in corneal neovascularization

膜 1 型基质金属蛋白酶在角膜新生血管形成中的新作用的鉴定

• 批准号: 10585794
• 负责人: Kyuyeon Han
• 金额: $ 39.98万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10585794
• 关键词: Angiogenesis Inhibitors; Angiogenic Factor; Angiostatins; Animal Model; Antibodies; Binding; Biological Assay; Biophysics; Blindness; Blood Vessels; Cells; Cellular Infiltration; Clioquinol; Cornea; Corneal Diseases; Corneal Injury; Corneal Neovascularization; Effectiveness; Endostatins; Endothelial Cells; Enzymes; Equilibrium; Event; Extracellular Matrix; Extracellular Matrix Degradation; FDA approved; FGF2 gene; FGFR2 gene; Fibroblast Growth Factor; Fibroblasts; Folic Acid; Gelatinase A; Gelatinase B; Goals; Impairment; Implant; In Vitro; Infection; Infiltration; Inflammation; Inflammatory; Injury; KDR gene; Knock-out; Knockout Mice; Lead; Libraries; MMP14 gene; Macrophage; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Measures; Mediating; Molecular Biology Techniques; Mus; Neutrophil Collagenase; Pharmaceutical Preparations; Proteins; Proteolysis; Regulation; Reporting; Role; Signal Transduction; Specificity; System; TNF-alpha converting enzyme; Testing; Therapeutic; Therapeutic Agents; Tissues; VEGFA gene; Validation; Vascular Endothelial Cell; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factors; Vision; Wild Type Mouse; analog; cell type; conditional knockout; cytokine; cytotoxicity; enzyme activity; experimental study; immune cell infiltrate; in vivo; inhibitor; injured; innovation; membrane activity; mutant; novel; novel therapeutics; pigment epithelium-derived factor; proMMP-2; protein expression; quinoline; quinoline analog; receptor; scaffold; side effect; small molecule; small molecule inhibitor

Project Summary/Abstract Corneal neovascularization (NV) can be caused by severe corneal injury or infection and is a leading cause of blindness. Balance of pro-angiogenic factors and anti-angiogenic factors are important to maintain avascular corneal tissue. Pro-angiogenic factors such as VEGFA and FGF2 are highly induced in inflamed corneas and lead to activation of its receptor proteins such as VEGFR2 and FGFR2. Membrane-type 1 matrix metalloproteinase (MMP-14) is involved in remodeling of extracellular matrix (ECM) through its proteolytic activity. Recent studies have revealed that MMP-14 is a regulator of VEGFA/VEGFR2-mediated corneal NV via unique and selective cleavage of VEGFR1 which is a decoy receptor for VEGFR2. FGF2-induced corneal NV is delayed in MMP-14 knockout (KO) mice, indicating there is some correlation between FGF2 and MMP-14. However, how MMP-14 interacts with FGF2 is still largely unknown. The goal of this application is to characterize mechanism of MMP-14 on regulation of FGFR2 levels via ADAM-9 enzyme. We demonstrated that FGFR2 level was low in MMP-14 KO corneal fibroblast cells. On the other hand, ADAM-9, which is substrate of MMP-14, was higher in MMP-14 KO fibroblast than WT cells. We have also discovered that expression of MMP-8, MMP-9, and ADAM-17, all of which are underlying FGF2/FGFR2-system, were highly induced in WT than MMP-14 KO cells upon stimulation of FGF2. Thus, inhibition of MMP-14 can reduce FGF2/FGFR2-mediated corneal NV and inflammation. Furthermore, our results show that FDA-approved small molecule drugs, clioquinol, chloroxine, and folic acid, all of which contain a quinoline scaffold, inhibit MMP-14 enzyme activity. We propose three specific aims: (1) Mechanism of two enzyme cascades, MMP-14 and ADAM-9, to regulate FGFR2 level and expression of FGF2/FGFR2-mediated MMPs; (2) investigate the effect of MMP-14 in FGF2/FGFR2-mediated corneal inflammation; (3) Characterize quinoline analogs as selective MMP-14 inhibitors. We will complete these aims using innovative techniques from molecular biology and biophysics in vitro and in vivo.

项目摘要/摘要 角膜新血管形成（NV）可能是由严重的角膜损伤或感染引起的，是主要原因 失明。促血管生成因子和抗血管生成因子的平衡对于维持血管很重要 角膜组织。在发炎的角膜和 导致其受体蛋白（例如VEGFR2和FGFR2）的激活。膜型1矩阵 金属蛋白酶（MMP-14）通过其蛋白水解对细胞外基质（ECM）进行重塑 活动。最近的研究表明，MMP-14是VEGFA/VEGFR2介导的角膜NV通过 VEGFR1的独特和选择性切割，这是VEGFR2的诱饵受体。 FGF2诱导的角膜NV是 MMP-14敲除（KO）小鼠的延迟，表明FGF2和MMP-14之间存在一些相关性。 但是，MMP-14与FGF2的相互作用仍然很大程度上未知。此应用的目的是表征 MMP-14通过ADAM-9酶调节FGFR2水平的机制。我们证明了FGFR2水平 在MMP-14 KO角膜成纤维细胞中较低。另一方面，MMP-14的基材Adam-9是 在MMP-14 KO成纤维细胞中，比WT细胞更高。我们还发现MMP-8，MMP-9和 ADAM-17（所有这些都是FGF2/FGFR2系统的基础）在WT中高于MMP-14 KO细胞高度诱导 刺激FGF2。因此，抑制MMP-14可以降低FGF2/FGFR2介导的角膜NV和 炎。此外，我们的结果表明，FDA批准的小分子药物，氯喹醇，氯氧醇， 和所有含有喹啉支架的叶酸抑制MMP-14酶活性。我们提出了三个特定的 目的：（1）两个酶级联的机制MMP-14和ADAM-9，以调节FGFR2水平和表达 FGF2/FGFR2介导的MMP; （2）研究MMP-14在FGF2/FGFR2介导的角膜中的影响 炎; （3）将喹啉类似物描述为选择性MMP-14抑制剂。我们将完成这些目标 利用体外和体内分子生物学和生物物理学的创新技术。