Functional and Integrative Omics of Recurrent Gout Flares

项目摘要 尿酸单钠晶体（MSU）激活炎性免疫细胞（如单核细胞/中性粒细胞）， 加重急性痛风发作。超声波和双能CT扫描显示存在MSU晶体 无症状痛风患者关节中的沉积物。这些观察使专家们同意，其他未知的 分子因素是痛风和复发性发作的触发因素。我们的辅助项目的目标是解决两个主要问题 问题：1）嗜中性粒细胞和/或单核细胞，在离体激活MSU晶体，表现出基因 复发性痛风发作的痛风患者之间的表达和DNA甲基化差异， 那些没有？2)在来自临床试验环境的痛风患者中， 在给予化疗（化疗）后，与外周血淋巴细胞相比， 有或没有复发性耀斑的个体之间的血液？在目标1中，采用机械/功能方法， 将使用新的痛风患者外周血新鲜样本分离中性粒细胞和CD 14+单核细胞 招募到UAB的痛风登记处，这是R 01的时间敏感的父项目。UAB痛风登记处是 由UAB的NIAMS资助的翻译研究中心（CORT）管理的“痛风研究， 高尿酸血症和合并症（INSIGHT）”（NIH P50 060772），这是我们辅助研究的父项目。 study.细胞将用MSU激活，我们将使用RNAseq比较细胞之间的基因表达。 组（复发性发作vs无发作）。我们将确定DNA甲基化变化是否与 观察到的转录差异。在aim 2中，我们将利用来自 临床试验，VA-CSP 594，“停止痛风”试验。该试验是一项非劣效性比较， 药物，非布司他和别嘌呤醇，主要结局， 一次停止痛风试验组织者同意增加他们的生物标本收集，以允许收集 我们的辅助研究。我们将分析来自试验参与者的约300份RNA样本。我们将 比较基线基因表达和DNA甲基化与复发性耀斑的相关性。我们将利用 孟德尔随机化，以评估差异甲基化CpG和表达转录物的因果关系 痛风复发目标1和目标2的结果将扩展我们对转录组学的认识， 甲基化的变化，沉淀痛风发作，这将转化为改善痛风患者的护理， 治疗

PROJECT SUMMARY Monosodium urate crystals (MSU) activate inflammatory immune cells (e.g. monocytes/neutrophils), which potentiate acute gout attacks. Ultrasound and dual energy CT scans have shown the presence of MSU crystal deposits in joints of asymptomatic gout patients. These observations lead experts to agree that other unknown molecular factors are triggers for gout and recurrent flares. Our ancillary project's aims address two main questions: 1) Do neutrophils and/or monocytes, upon ex vivo activation by MSU crystals, exhibit gene expression and DNA methylation differences between gout patients with recurrent gout flares compared to those that do not? 2) Among gout patients from a clinical trial setting, where treat-to-target urate lowering therapy (ULT) is administered, are there gene expression and DNA methylation differences from the peripheral blood between individuals with or without recurrent flares? In aim 1 in a mechanistic/functional approach we will isolate neutrophils and CD14+ monocytes using fresh samples of peripheral blood from gout patients newly recruited into UAB's Gout Registry, the time sensitive parent project for this R01. The UAB Gout Registry is administered by UAB's NIAMS funded Center of Research Translation (CORT) “INvestigationS In Gout, Hyperuricemia, and comorbidiTies (INSIGHT)” (NIH P50 060772), which is the parent project for our ancillary study. The cells will be activated with MSU and we will use RNAseq to compare gene expression between groups (recurrent flares vs no flares). We will determine whether DNA methylation changes may be associated with observed transcription differences. In aim 2 we will utilize clinical data and biological samples from the clinical trial, VA-CSP594, the “Stop Gout” trial. The trial is a non-inferiority comparison between the ULT agents, febuxostat and allopurinol, of the primary outcome, the proportion of participants who flare at least once. The Stop Gout trial organizers agreed to augment their biospecimen collection to allow the collection of RNA for our ancillary study. We will analyze ~300 samples of RNA from the trial's participants. We will compare baseline gene expression and DNA methylation associations with recurrent flares. We will utilize Mendelian randomization to assess the causality of differentially methylated CpGs and expressed transcripts with recurrent gout flares. The results from aims 1 and 2 will expand our knowledge of the transcriptomic and methylomic changes that precipitate gout flares, which will translate to improved gout patient care and treatment.