PROTEINASE ACTIVATED RECEPTORS IN CORNEAL INFLAMMATION

角膜炎症中的蛋白酶激活受体

• 批准号: 6668508
• 负责人: JOHN C ANSEL
• 金额: $ 14.61万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6668508
• 关键词: cell differentiation; cell migration; cell proliferation; clinical research; collagenase; cornea edema; corneal epithelium; enzyme activity; flow cytometry; human tissue; immunocytochemistry; inflammation; laboratory mouse; receptor expression; serine proteinases; tissue inhibitor of metalloproteinases; wound healing

DESCRIPTION (provided by applicant): Proteinase activated receptors (PARs) comprise a novel family of cell membrane proteins that are potent mediators of tissue inflammation, pain, and repair. PARs are activated by serine proteases such as thrombin, trypsin, and tryptase that are released in injured or inflamed tissues. We have recently reported that the cornea expresses two of the principal proteinase activated receptors: PAR-1 and PAR-2. The long-term goal of this project is to determine the role of serine proteases and PARs to mediate a variety of corneal epithelial cell wound healing and inflammatory responses. This application will test the hypothesis that serine proteases such as thrombin and tryptase are potent modulators of corneal epithelial cell inflammatory and wound healing responses that are mediated by the activation of specific corneal epithelial cell PARs. In this proposal, we will undertake the following specific Aims: SPECIFIC AIM #1: To examine the effect of seine proteases on human corneal epithelial cell biological activities critical for corneal inflammatory and wound healing responses; SPECIFIC AIM #2: To determine if corneal epithelial cell responses to serine proteases are mediated by the activation of specific functional protease activated receptors (PARs); SPECIFIC AIM #3: To determine the role of serine protease activation of PARs in mediating in vivo corneal inflammatory responses. To conduct the proposed studies, human corneal epithelial cells and human corneal tissue will be utilized for in vitro studies and the proposed in vivo studies will utilize both normal mice as well as PAR-1 and PAR-2 knockout mice. The ability of serine proteases to influence human corneal epithelial cell proliferation, differentiation, migration, MMP/TIMP expression, and proinflammatory protein production will be determined in vitro. The specific role of PAR-1 and PAR-2 in these serine protease induced human corneal epithelial cell responses will be assessed using serine proteases and I specific native peptide derived PAR-1 and PAR-2 agonists and antagonists. Additionally, preliminary "proof of principle" in vivo studies will be undertaken to determine the capacity of serine proteases and specific PAR-1 and PAR-2 agonists to induce corneal inflammatory responses in wild type mice in comparison to PAR-I(-/-) and I PAR-2 (-/-) animals. Proteases and PARs may be important mediators of corneal inflammation and wound healing and therefore could prove to be novel therapeutic targets for a wide range of ocular diseases.

描述（由申请人提供）：蛋白酶激活受体（PARs）包括一个新的细胞膜蛋白家族，它们是组织炎症、疼痛和修复的有效介质。PAR被丝氨酸蛋白酶激活，如凝血酶、胰蛋白酶和类胰蛋白酶，它们在受伤或发炎的组织中释放。我们最近报道，角膜表达两种主要的蛋白酶激活受体：PAR-1和PAR-2。本项目的长期目标是确定丝氨酸蛋白酶和PAR在介导各种角膜上皮细胞伤口愈合和炎症反应中的作用。本申请将检验丝氨酸蛋白酶如凝血酶和类胰蛋白酶是角膜上皮细胞炎症和伤口愈合反应的有效调节剂的假设，所述炎症和伤口愈合反应由特定角膜上皮细胞PAR的活化介导。在本提案中，我们将承担以下具体目标：具体目标#1：检查丝氨酸蛋白酶对角膜炎症和伤口愈合反应关键的人角膜上皮细胞生物活性的影响;具体目标#2：确定角膜上皮细胞对丝氨酸蛋白酶的反应是否由特定功能性蛋白酶激活受体（PAR）的激活介导;具体目标#3：确定PAR的丝氨酸蛋白酶活化在介导体内角膜炎症反应中的作用。为了进行拟定的研究，将使用人角膜上皮细胞和人角膜组织进行体外研究，拟定的体内研究将使用正常小鼠以及PAR-1和PAR-2敲除小鼠。将在体外测定丝氨酸蛋白酶影响人角膜上皮细胞增殖、分化、迁移、MMP/TIMP表达和促炎蛋白产生的能力。PAR-1和PAR-2在这些丝氨酸蛋白酶诱导的人角膜上皮细胞应答中的特定作用将使用丝氨酸蛋白酶和I特异性天然肽衍生的PAR-1和PAR-2激动剂和拮抗剂来评估。此外，将进行初步的“原理证明”体内研究，以确定丝氨酸蛋白酶和特异性PAR-1和PAR-2激动剂与PAR-1（-/-）和PAR-2（-/-）动物相比在野生型小鼠中诱导角膜炎症反应的能力。蛋白酶和PAR可能是角膜炎症和伤口愈合的重要介质，因此可能被证明是广泛的眼部疾病的新的治疗靶点。