Role of the Novel Matrix Metalloproteinase Epilysin in Atopic Dematitis

描述（由申请人提供）：a . Epilysin （matrix metalloproteinase [MMPJ-28）是一种由角质形成细胞和其他上皮细胞类型组成表达的新型MMP。迄今为止，对脱毛素的生理底物和体内功能知之甚少。我们现在有证据表明，癫痫素在调节th2型过敏性炎症中的潜在作用，如哮喘和特应性皮炎（AD）。在哮喘和AD小鼠模型中（分别基于气道和表皮卵清蛋白挑战），癫痫素在急性炎症期间下调，然后随着时间的推移而增加。此外，与野生型小鼠相比，缺乏癫痫素的小鼠分别经历了更多的肺部和皮肤炎症。脱毛素也被TNF-a上调，TNF-a是过敏性炎症的重要介质。基于这些观察结果，我们假设癫痫素通过影响控制炎症的蛋白质活性在特应性炎症中发挥保护作用，并且其表达受炎症细胞因子环境的调节。AD是一种常见的慢性皮肤疾病，其发病机制和调节尚不完全清楚，因此缺乏针对该疾病的治疗方法。我们的长期目标是阐明和定义阿尔茨海默病发病机制的潜在关键调控方面，即脱毛素的作用，从而确定疾病干预的前瞻性位点。为此，我们建议定义癫痫素在炎症中的作用，它的时间表达和调控。我们将使用AD小鼠模型来评估炎症期间癫痫素的时间表达。在特定的时间点，我们将通过RT-PCR测定癫痫素的表达，并将其与组织学炎症程度、血清IgE、细胞和细胞因子表型相关联。然后，我们将使用已建立的无脱毛素小鼠来比较上述炎症表型与没有脱毛素假定的调节功能的小鼠。同时，我们将研究体外调控方面。我们将在不同炎症细胞因子处理的培养角化细胞中检测epilysin的表达。我们还将采用荧光素酶测定与已建立的启动子结构进一步表征假定的启动子调控域。B.特应性皮炎（AD）影响多达20%的学龄儿童，并具有显著的身体、经济和社会心理影响。通过研究一种似乎在AD中发挥保护作用的分子，我们的目标是增加我们对AD如何调节的认识，并为未来的新疗法提供可能性。

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.