Investigation of Nod2: Implications for Uveitis

描述(申请人提供)：葡萄膜炎的潜在机制仍然知之甚少。然而，最近发现的一种名为NOD2的基因，为了解葡萄膜炎发病的遗传学机制提供了重要的见解。NOD2基因突变导致Blau综合征和易患克罗恩病，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，这项建议试图研究NOD2在眼睛中的表达，并研究与疾病相关的NOD2突变如何改变细胞炎症功能。虽然NOD2最初被认为主要在单核细胞中表达，但提供的初步数据表明，NOD2分布更广泛，似乎位于受BLAU综合征影响的主要组织-眼睛。Aim1的重点是证明NOD2位于眼睛内。NOD2的细胞分布将通过免疫组织化学方法在人眼组织中进行检测。我们将在培养的人虹膜/脉络膜组织的内皮细胞中检测NOD2的调节，我们将测试内皮细胞对NOD2配体(MDP)处理的功能性反应。在AIM2中，我们将使用转染的细胞系来研究在MDP启动的背景下，异常的NOD2(由于BLAU突变(R334Q)或克罗恩突变(L1007fs))如何改变NF-kB的活性和细胞因子的产生，以响应MDP处理或TLR反应。总之，这些研究将为NOD2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.