Regulation of Inflammation by IL1RN polymorphisms

IL1RN 多态性对炎症的调节

• 批准号: 10359078
• 负责人: Steven B Abramson
• 金额: $ 18.46万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10359078
• 关键词: Affect; Biological Assay; Chronic Childhood Arthritis; Clinical; Degenerative polyarthritis; Development; Diabetes Mellitus; Disease; Disease model; ES Cell Line; Elements; Event; Gene Expression Regulation; Genes; Genetic; Genetic Polymorphism; Genotype; Grant; Haplotypes; High Fat Diet; Human; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Insulin Resistance; Interleukin Gene; Joints; Kellgren-Lawrence grade; Knee Osteoarthritis; Knock-out; Length; Link; Literature; Medial; Medial meniscus structure; Methods; Modeling; Molecular; Mus; Obesity; Operative Surgical Procedures; Outcome; Pathogenesis; Patients; Phenotype; Production; Regulation; Rheumatoid Arthritis; Risk; Role; Severities; Severity of illness; Surface; Technology; Testing; Transgenic Mice; Transgenic Organisms; Width; age related; anakinra; cytokine; diet-induced obesity; disorder prevention; disorder risk; embryonic stem cell; gene regulatory network; genomic locus; human disease; human model; humanized mouse; in vivo; in vivo evaluation; macrophage; monocyte; mouse model; novel; obese patients; patient population; radiological imaging; response; risk variant; screening

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.

抽象的 有重要文献表明 IL1RN 多态性影响多种疾病的临床结果， 包括骨关节炎、糖尿病、幼年关节炎和肥胖症 传递 IL1RN 的精确机制。 我们已经证明 IL1RN TTG 单倍型（rs419598、rs315952 和 rs9005）可以预测。 与年龄相关的放射线严重程度 [Kellgren-Lawrence 分级 (KL)、内侧关节间隙宽度 (JSW)] 和 此外，这种促炎基因型与膝关节 OA 的风险增加 3 倍无关。 仅限于 OA，但可以在类风湿性关节炎 (RA) 患者和肥胖患者中得到证实 IL1RN TTG 风险单倍型 IL1RN 基因中有许多功能元件，目前尚不清楚。 其中哪一个通过调节 IL1RN 表达并影响其他因素来增强炎症功能 因此，这项资助的中心目标是确定单倍型块中的基因的影响。 使用转基因技术在体外和体内检测 IL1RN 风险单倍型 目标 1：使用合成人类。 单倍型以确定哪些功能元件对于炎症表型至关重要 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单倍型。 人类基因使用“Assemblon”技术，可以构建和精确传递哺乳动物的基因 使用该技术，我们将选择长度高达200 kb的基因位点信息丰富的小鼠胚胎干细胞。 (mESC) 系，并将在体外分化为单核细胞和巨噬细胞，并对其进行分析 IL1RN 和单倍型块中其他基因的表达、细胞因子的产生、表面标记 目标 2：生成表达 IL1RN TTG-2 风险或 TTG-0 非风险的人源化小鼠。 用于研究手术诱导的 OA 内侧半月板 (DMM) 模型不稳定的单倍型。 最近的研究还表明 IL1RN 基因多态性与肥胖和胰岛素抵抗之间存在关联，我们 将在转基因人源化小鼠中研究高脂肪饮食诱导的肥胖模型，这些研究将阐明新的方法。 IL1RN 风险单倍型影响细胞反应和炎症基因调控的机制， 并将揭示以前未被识别的各种功能相关基因，这些基因对于炎症至关重要 风险单倍型的功能。