Regulation of Inflammation by IL1RN polymorphisms

摘要 有重要的文献表明，IL1RN基因多态性会影响多种疾病的临床结果， 包括骨性关节炎、糖尿病、青少年关节炎和肥胖症。传递IL1RN的精确机制 影响尚不清楚。我们已经证明IL1RN TTG单倍型(rs419598、rs315952和rs9005)预测 与年龄相关的放射学严重程度[Kellgren-Lawrence分级(KL)、内侧关节间隙宽度(JSW)]和IS 与发生膝关节骨关节炎的风险增加3倍相关。此外，这种促炎基因并不是 仅限于骨性关节炎，但可见于类风湿性关节炎(RA)患者和携带 IL1RN TTG风险单倍型。IL1RN基因中有许多功能元件，目前尚不清楚 其中哪一种通过调节IL1RN的表达和影响其他 单倍型中的基因块。因此，这笔赠款的主要目标将是界定 使用转基因技术在体外和体内进行IL1RN风险单倍型研究。目标1：使用人造人类 单倍型以确定哪些功能元件对由 IL1RN风险单倍型，体外。我们将在侧翼的背景下构建风险和保护性单倍型 人类基因使用“组装”技术，这使得哺乳动物的构建和精确交付 长达200kb的基因座。利用这项技术，我们将选择信息丰富的小鼠胚胎干细胞 (MESC)，并将在体外分化为单核细胞和巨噬细胞，将被检测 白介素1RN等基因在单倍型阻断中的表达、细胞因子产生、表面标记 表情。目的2：建立表达IL1RN TTG-2风险或TTG-0非风险的人源化小鼠 单倍型在手术诱导内侧半月板(DMM)骨关节炎模型中的研究。自.以来 最近的研究还表明，IL1RN基因多态与肥胖和胰岛素抵抗有关 将在转基因人源化小鼠中研究高脂饮食诱导的肥胖模型。这些研究将阐明小说 IL1RN风险单倍型影响细胞反应和炎症基因调控的机制 并将揭示以前未知的各种功能相关基因，这些基因对炎症性疾病至关重要 风险单倍型的功能。

ABSTRACT There is significant literature that IL1RN polymorphisms affect clinical outcomes in a variety of diseases, including osteoarthritis, diabetes, juvenile arthritis and obesity. The precise mechanisms that impart IL1RN's effects are unknown. We have shown that the IL1RN TTG haplotype (rs419598, rs315952 and rs9005) predicts both age-related radiographic severity [Kellgren-Lawrence grade (KL), medial joint-space width (JSW)] and is associated with a 3-fold increased risk for incident knee OA. Moreover, this pro-inflammatory genotype is not limited to OA but can be demonstrated in patients with rheumatoid arthritis (RA) and in obese patients who carry the IL1RN TTG risk haplotype. There are a number of functional elements in the IL1RN gene, and it is unclear which of these confers increased inflammatory function by modulating IL1RN expression and influencing other genes in the haplotype block. The central objective of this grant, therefore, will be to define the effects of the IL1RN risk haplotypes in vitro and in vivo using transgenic technologies. In Aim 1: Use synthetic human haplotypes to determine which functional elements are critical for the inflammatory phenotype conferred by the IL1RN risk haplotype, in vitro. We will construct risk and protective haplotypes in the context of the flanking human genes using “Assemblon” technology, which allows the construction and precision delivery of mammalian gene loci of up to 200 kb in length. Using this technology, we will select informative mouse embryonic stem cell (mESC) lines and will be differentiated in vitro to monocytes and macrophages which will be assayed for expression of IL1RN and the other genes in the haplotype block, cytokine production, surface marker expressions. Aim 2: Generate a humanized mouse expressing the IL1RN TTG-2 risk or TTG-0 non-risk haplotypes for study in the surgically induced destabilization of the medial meniscus (DMM) model of OA. Since recent studies also indicate an association of IL1RN genetic polymorphism in obesity and insulin resistance, we will study high-fat diet-induced obesity models in transgenic humanized mice. These studies will elucidate novel mechanisms by which the IL1RN risk haplotype influences cellular responses and inflammatory gene regulation, and will reveal previously unrecognized various functional linked genes, which are crucial for the inflammatory function of the risk haplotype.