Role of the Novel Matrix Metalloproteinase Epilysin in Atopic Dematitis

说明书(申请人提供)：a.基质金属蛋白酶[MMPJ-28]是一种由角质形成细胞和其他上皮细胞成分表达的新型基质金属蛋白酶。到目前为止，对外膜粘附素的生理底物(S)及其在体内的作用知之甚少。我们现在有证据表明，Eepysin在调节Th2型过敏性炎症方面具有潜在作用，就像哮喘和特应性皮炎(AD)中发生的那样。在哮喘和AD小鼠模型中(分别基于呼吸道和皮肤表面卵白蛋白攻击)，在急性炎症期间，Eepysin下调，然后随着时间的推移似乎增加。此外，与受到挑战的野生型小鼠相比，缺乏羽毛粘连蛋白的小鼠的肺部和皮肤炎症分别增加。过敏性炎症的重要介质--肿瘤坏死因子-a也上调了表皮溶血素。基于这些观察，我们假设Eepysin在特应性炎症过程中通过影响控制炎症的蛋白质的活性而发挥保护作用，并且它的表达受炎症细胞因子环境的调节。AD是一种常见的、慢性的、经常使人衰弱的皮肤病，其发病机制和调节机制尚不完全清楚，因此缺乏针对该疾病的特异性治疗方法。我们的长期目标是阐明和定义AD发病机制中一个潜在的关键调控方面，即Eepysin的作用，从而确定疾病干预的预期位置。为此，我们建议定义外切蛋白在炎症中的作用，它的时间表达，以及它的调节。我们将使用阿尔茨海默病的小鼠模型来评估炎症过程中Eepysin的瞬时表达。在特定的时间点，我们将通过RT-PCR检测Eepysin的表达，并将其与每一组织学的炎症程度、血清IgE以及细胞和细胞因子表型相关联。然后，我们将使用已建立的Eepysin缺失的小鼠来比较上述炎症表型与缺乏Eepysin的假定调节功能时所见的炎症表型。同时，我们将检查体外调控方面的情况。我们将检测不同炎性细胞因子处理的培养角质形成细胞中Eepysin的表达。我们还将利用已建立的启动子结构的荧光素酶分析来进一步表征假定的启动子调节结构域。B.特应性皮炎(AD)影响多达20%的学龄儿童，并对身体、经济和心理社会产生重大影响。通过研究一种似乎在阿尔茨海默病中发挥保护作用的分子，我们的目标是增加我们对阿尔茨海默病是如何调节的知识，以及未来新疗法的可能性。

DESCRIPTION (provided by applicant): A. Epilysin (matrix metalloproteinase [MMPJ-28) is a novel MMP expressed constitutively by keratinocytes and other epithelial cell types. To date, little is known about epilysin's physiologic substrate(s) and in vivo function. We now have evidence suggesting a potential role for epilysin in modulating Th2-type allergic inflammation, as occurs in asthma and atopic dermatitis (AD). In both asthma and AD mouse models (based on airway and epicutaneous ovalbumin challenge, respectively), epilysin is downregulated during acute inflammation and then appears to increase over time. Furthermore, mice lacking epilysin experience increased lung and skin inflammation, respectively, compared to challenged wild-type mice. Epilysin is also upregulated by TNF-a, an important mediator of allergic inflammation. Based on these observations, we hypothesize that epilysin plays a protective role during atopic inflammation by affecting the activity of proteins that govern inflammationand that its expression is regulated by an inflammatory cytokine milieu. AD is a common, chronic, and often debilitating skin disease whose pathogenesis and regulation are still incompletely understood and for which disease-specific therapeutics are therefore lacking. Our long-term objective is to elucidate and define a potentially critical regulatory aspect of AD pathogenesis, namely the role of epilysin, thereby identifying a prospective site for disease intervention. To this end,we propose to define epilysin's role in inflammation, its temporal expression, and its regulation. We will use a mouse model of AD to evaluate the temporal expression of epilysin during inflammation. At particular time points, we will determine epilysin expression by RT-PCR and correlate this with degree of inflammation per histology, serum IgE,and cellular and cytokine phenotypes. We will then use established epilysin-null mice to compare the above inflammatory phenotypes to those seen in the absence of epilysin's putative regulatory function. Concurrently, we will examine regulatory aspects in vitro. We will examine epilysin expression in cultured keratinocytes treated with various inflammatory cytokines. We will also employ the luciferase assay with established promoter constructs to further characterize putative promoter regulatory domains. B. Atopic dermatitis (AD) affects up to 20% of school-aged children and has significant physical, economic, and psychosocial impact. By studying a molecule that appears to play a protective role in AD, we aim to increase our knowledge of how AD is regulated and the possibility for novel therapies in the future.