Caspase-14 and the Treatment of Psoriasis

描述(申请人提供)：牛皮癣是一种慢性皮肤病，以发炎、鳞屑和经常毁容的皮肤损害为特征。皮损表现为过度增殖和终末分化改变，导致角化不全和脱屑。Caspase-14是一个非凋亡性Caspase家族成员，参与终末分化，对屏障破坏后角化的加速和正常角膜细胞的形成是必不可少的。新出现的数据和我们令人信服的初步研究表明，与相同个体的非皮损皮肤和正常对照个体的相应样本相比，caspase-14在银屑病皮损中的表达显著下调。因此，识别具有诱导终末分化和抑制过度增殖能力的天然抗炎药可能对银屑病的治疗有用。飞雀素是一种主要的花青素，广泛存在于有色水果和蔬菜中，具有抗炎和抗增殖活性。我们未发表的初步研究值得注意的是，我们已经证明了Delphinidin治疗诱导了正常人表皮角质形成细胞(NHEK)中前天冬氨酸氨基转移酶-14的蛋白和mRNA表达。Delphinidin还诱导caspase-14被加工成具有催化活性的亚基p10和p20。我们还发现，在Delphinidin处理的NHEK中，总蛋白和转谷氨酰胺酶-1的蛋白和mRNA的表达显著增加。此外，Delphinidin处理NHEK后，AP-1亚基和NF-：B亚基p50和RelB的蛋白表达增加。重要的是，在相同的处理条件下，Delphinidin不会导致细胞凋亡。这种显著的差异构成了这一建议的基础，该建议旨在研究Delphinidin在体外人类重组皮肤模型和体内临床前环境中对角质形成细胞分化和过度增殖的影响。在这项建议中要检验的假设是“Delphinidin将诱导分化并加速角化，这反过来将通过诱导caspase-14的表达和在不诱导细胞凋亡的情况下抑制细胞增殖来减轻银屑病样皮损的严重程度”。为了验证我们的假设，我们提出了以下具体目标：(I)研究在三维人体重建皮肤模型中，飞蓬毒素治疗是否通过增加caspase-14的表达和处理来加速角化过程，(Ii)研究在三维人体重建皮肤模型中，飞虱对caspase-14表达、细胞定位和处理的影响是否通过AP-1和NF-：B通路介导。以及(Iii)通过使用片状皮肤(FSN/FSN)小鼠和片状皮肤(FSN/FSN)caspase 14-/-小鼠，在活体情况下，确定caspase-14是否与Delphinidin治疗的表皮病理症状减轻有关。这项提案将确定caspase-14在银屑病治疗中的作用，此外还将提请注意使用一种花青素--飞雀素来治疗银屑病。

DESCRIPTION (provided by applicant): Psoriasis is a chronic skin disease characterized by inflamed, scaly and frequently disfiguring skin lesions. The skin lesions show hyperproliferation and altered terminal differentiation leading to parakeratosis and desquamation. Caspase-14 is a nonapoptotic caspase family member and is involved in terminal differentiation and is essential for accelerated cornification in response to barrier disruption and for the formation of normal corneocytes. Emerging data and our compelling preliminary studies suggest that caspase-14 expression is substantially down-regulated in human psoriatic lesions compared to corresponding samples from nonlesional skin of the same individuals and from normal control individual. Thus, identifying naturally occurring anti-inflammatory agents which possess the ability to induce terminal differentiation and inhibit hyperproliferation could be useful for the treatment of psoriasis. Delphinidin, a major anthocyanidin abundantly present in pigmented fruits and vegetables, possesses anti-inflammatory and anti-proliferative activities. Our preliminary unpublished studies are noteworthy where we have demonstrated that delphinidin treatment induced the protein and mRNA expression of procaspase-14 in normal human epidermal keratinocytes (NHEK). Delphinidin also induced the processing of caspase-14 into catalytically active subunits p10 and p20. We also found a significant increase in the protein and mRNA expression of involucrin and transglutaminase-1 in delphinidin treated NHEK. Furthermore, delphinidin treatment to NHEK increased the protein expression of AP-1 subunits and NF-:B subunits p50 and RelB. Importantly, delphinidin under identical treatment conditions did not result in induction of apoptosis. This remarkable distinction forms the basis of this proposal which is designed to investigate the effect of delphinidin on keratinocyte differentiation and hyperproliferation both in in vitro human reconstituted skin model and in preclinical in vivo settings. The hypothesis to be tested in this proposal is that "delphinidin will induce differentiation and accelerate cornification that will in turn reduce the severity of psoriasiform lesions by inducing the expression of caspase-14 and suppression of cell proliferation without inducing apoptosis". To test our hypothesis, the following specific aims are proposed: (i) To investigate whether delphinidin treatment accelerates the process of cornification through increased expression and processing of caspase-14 in three-dimensional human reconstituted skin model, (ii) To investigate whether the effect of delphinidin on caspase-14 expression, cellular localization, and processing is mediated through AP-1 and NF-:B pathways in three-dimensional human reconstituted skin model, and (iii) To establish whether caspase-14 is associated with reduced symptoms of epidermal pathology with delphinidin treatment under in vivo situation by employing flaky skin (fsn/fsn) mice and flaky skin (fsn/fsn) caspase 14-/- mice. This proposal will establish the role of caspase-14 for treatment of psoriasis and in addition will draw attention to the use of delphinidin an anthocyanidin for its treatment.