Regulation of Inflammation by IL1RN polymorphisms

摘要 有大量文献表明IL 1 RN多态性影响多种疾病的临床结果， 包括骨关节炎、糖尿病、青少年关节炎和肥胖症。传递IL 1 RN的精确机制 影响未知。我们已经表明，IL 1 RN TTG单倍型（rs 419598，rs315952和rs 9005）预测 与年龄相关的放射学严重程度[Kelling-Lawrence分级（KL），内侧关节间隙宽度（JSW）]， 与发生膝关节OA的风险增加3倍相关。此外，这种促炎基因型不是 局限于OA，但可在类风湿性关节炎（RA）患者和携带 IL 1 RN TTG风险单倍型。IL 1 RN基因中有许多功能元件，目前尚不清楚 这些通过调节IL 1 RN表达和影响其他细胞因子而赋予增加的炎症功能， 单倍型块中的基因。因此，这笔赠款的中心目标是确定 使用转基因技术在体外和体内研究IL 1 RN风险单倍型。目标1：使用人造人 单倍型，以确定哪些功能元件是关键的炎症表型赋予的 IL 1 RN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“装配”技术，这使得建设和精确交付哺乳动物 基因座长度可达200 kb。利用该技术，我们将筛选出有信息的小鼠胚胎干细胞， 细胞系，并将在体外分化为单核细胞和巨噬细胞， IL 1 RN和其他基因在单倍型块中的表达，细胞因子产生，表面标记物 表情目的2：产生表达IL 1 RN TTG-2风险或TTG-0非风险的人源化小鼠 单倍型用于OA的手术诱导的内侧半月板（DMM）不稳定模型的研究。以来 最近的研究也表明IL 1 RN基因多态性与肥胖和胰岛素抵抗有关，我们 将在转基因人源化小鼠中研究高脂饮食诱导的肥胖模型。这些研究将阐明新的 IL 1 RN风险单倍型影响细胞反应和炎症基因调节的机制， 并将揭示以前未被识别的各种功能相关基因，这些基因对炎症反应至关重要。 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.