Investigation of Nod2: Implications for Uveitis

描述（由申请人提供）：葡萄膜炎的潜在机制仍然知之甚少。然而，最近发现的一个称为Nod 2的基因为葡萄膜炎发病机制中涉及的遗传学提供了重要的见解。Nod 2的突变导致Blau综合征并易患克罗恩病，这两种疾病都是以葡萄膜炎为特征的自身免疫性疾病。因此，该提案旨在研究Nod 2在眼睛中的表达，并研究Nod 2中的疾病相关突变如何改变细胞炎症功能。虽然Nod 2最初被认为主要在单核细胞中表达，但初步数据表明Nod 2更广泛，似乎位于眼睛中，这是Blau综合征的主要组织。Aim 1的重点是证明Nod 2位于眼睛中。将通过免疫组织化学在人眼外植体中检查Nod 2的细胞分布。将在来自人虹膜/脉络膜组织的培养的内皮细胞中检查Nod 2调节，并且我们将测试内皮培养物是否在功能上响应于用Nod 2配体（MDP）处理。在Aim 2中，我们将使用转染的细胞系来研究异常的Nod 2（由于Blau突变（R334 Q）或Crohn突变（L1007 fs））如何改变NF-kB活性和细胞因子产生以响应MDP处理或MDP引发设置中的TLR响应。总之，这些研究将为Nod 2如何参与自身免疫和葡萄膜炎的发病机制提供有价值的新信息。

DESCRIPTION (provided by applicant): The underlying mechanisms of uveitis remain poorly understood. However, a recently identified gene called Nod2, provides important insight into the genetics involved in the pathogenesis of uveitis. Mutations in Nod2 result in Blau syndrome and predispose to Crohn's disease, both of which are autoimmune diseases characterized by uveitis. Thus, this proposal seeks to investigate Nod2 expression in the eye and examine how disease-associated mutations in Nod2 alter cellular inflammatory functions. Although Nod2 was originally thought to be primarily expressed in monocytes, the preliminary data presented suggest that Nod2 is more widespread and appears to be located in the eye, the main tissue affected by Blau syndrome. The focus of Aim1 is to demonstrate the Nod2 is located in the eye. The cellular distribution of Nod2 will be examined by immunohistochemistry in human eye explants. Nod2 regulation will be examined in cultured endothelilal cells from human iris/choroid tissue, and we will test whether endothelial cultures functionally respond to treatment with Nod2-ligand (MDP). In Aim2 we will use transfected cell lines to investigate how abnormal Nod2 (due to Blau-mutation (R334Q) or Crohn's-mutation (L1007fs)) alters NF-kB activity and cytokine production in response to MDP-treatment or TLR-responses in the setting of MDP-priming. Together, these studies will provide valuable new information on how Nod2 is involved in autoimmunity and the pathogenesis of uveitis.