Research Project II - Vesicant-Induced Corneal Injury

研究项目2.糜烂性角膜损伤 关键人员： 马里恩·戈登博士罗格斯大学药学院副教授 项目总结/摘要 硫芥（SM）和氮芥（NM）会对角膜上皮细胞造成微泡损伤， 基质缘当广泛时，上皮层脱落，导致长期后果，如 复发性角膜糜烂和角膜缘干细胞缺乏。我们的实验室一直在用兔子的角膜 器官培养，以分析水泡剂诱导的角膜损伤的潜在机制。我们已经发现 芥子气暴露后的角膜损伤是由于基质金属蛋白酶（MMPs）的激活， 降解基底膜区的细胞外基质，导致上皮层分离 从基质;这与延迟和有缺陷的伤口愈合有关。MMP的主要诱导剂 产生的是EMMPRIN（CD 147），我们发现其在暴露于发泡剂后在角膜中上调。 已知EMMPRIN是亲环蛋白A的受体，亲环蛋白A是具有促炎活性的细胞内蛋白， 从细胞中释放出来。我们假设芥子气暴露后的损伤是由于 亲环素A诱导的炎症和EMMPRIN引起的MMP活化。此外， 芥子气暴露的角膜是由于临时基质的延迟去除，以及 纤连蛋白。为了验证这些假设，我们将分析EMMPRIN和亲环蛋白在MMP激活中的作用。 在芥子气暴露后的伤口愈合中的作用使用一种抑制EMMPRIN作用的方法， 封闭抗体和使用环孢菌素A（CsA）的亲环素A将被评估。更好的治疗将是 通过减少上皮-基质分离、减少MMP 9活化和减少IL-1 表情我们还将确定芥子气暴露后角膜伤口愈合时间延长是否是由于 纤维连接蛋白（FN）的延迟沉积，这是支持细胞迁移以闭合损伤所需的。的 已知FN肽通过促进迁移而促进角膜上皮伤口愈合的能力， 分析了这些研究结果将提供重要的机制信息， 损伤，并可能导致治疗芥子损伤的新疗法的发展。

Research Project 2. Vesicant-Induced Corneal Injury Key Personnel: Marion Gordon, Ph.D., Associate Professor, Rutgers University School of Pharmacy Project Summary/Abstract Sulfur mustard (SM) and nitrogen mustard (NM) cause microblistering injury to the cornea at the epithelial- stromal border. When extensive, the epithelial layer sloughs off leading to long term consequences such as recurrent corneal erosions and limbal stem cell deficiency. Our laboratories have been using a rabbit cornea organ culture to analyze mechanisms underlying vesicant-induced injury to the cornea. We have discovered that corneal injury after mustard exposure is due to activation of matrix metalloproteinases (MMPs), which degrade the extracellular matrix of the basement membrane zone leading to separation of the epithelial layer from the stroma; this is associated with delayed and defective wound healing. A major inducer of MMP production is EMMPRIN (CD147), which we found is upregulated in the cornea following vesicant exposure. EMMPRIN is known to be a receptor for cyclophillin A, an intracellular protein with proinflammatory activity, released from cells following injury. We hypothesize that injury following mustard exposure is due to cyclophillin A-induced inflammation and MMP activation by EMMPRIN. Furthermore, slower wound healing of mustard-exposed corneas is due to delayed removal of provisional matrix, as well as delayed deposition of fibronectin. To test these hypotheses, we will analyze the role of EMMPRIN and cyclophillin in MMP activation in wound healing following mustard exposure. The effects of inhibiting the action of EMMPRIN using a blocking antibody, and cyclophillin A using cyclosporine A (CsA) will be assessed. Improved healing will be determined by a decrease in epithelial-stromal separation, decreased MMP9 activation and decreased IL-1 expression. We will also determine if prolonged corneal wound healing after mustard exposure is due to delayed deposition of fibronectin (FN) which is required to support the cells' migration to close the injury. The ability of a FN peptide known to promote corneal epithelial wound healing by facilitating migration will be analyzed. Results of these studies will provide important mechanistic information on vesicant induced corneal injury and may lead to the development of new therapies for treating mustard injury.