Department of Rheumatology, Hwa Mei Hospital, University of Chinese Academy of Sciences (Ningbo No. 2 Hospital), Ningbo, Zhejiang, China.
Medical School, Ningbo University, Ningbo, Zhejiang, China.
J Clin Lab Anal. 2022 Sep;36(9):e24634. doi: 10.1002/jcla.24634. Epub 2022 Jul 31.
The purpose of this study was to identify biomarkers for the diagnosis of gout in Chinese Han males using methylation microarray profiling.
We screened for differentially methylated genes (DMGs) in gout using a methylation microarray and analyzed the functions of the DMGs using gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. We verified gene methylation levels by pyrosequencing and protein levels by enzyme-linked immunosorbent assays (ELISAs). Statistical analyses were performed using SPSS. Two-sided p values <0.05 were deemed to be statistically significant for all analyses.
We identified 20,426 significant differential methylation sites (5719 high-methylation sites and 14,707 low-methylation sites). Bioinformatics analysis showed that the DMGs were mainly involved in 43 biological functions, 13 cellular components, 18 molecular functions, and 35 KEGG pathways. We selected opioid receptor delta 1 (OPRD1) for verification of methylation levels between 50 gout patients and 50 controls. The methylation levels of OPRD1 (Chr1:29,139,121) were significantly lower in the gout group (p < 0.05), while OPRD1 protein levels were significantly higher in the gout group (p < 0.05). In addition, the AUC of the combination of OPRD1 (Chr1:29,139,121) methylation and OPRD1 protein levels was 0.796 (0.710, 0.883) with a high sensitivity of 82% and a specificity of 68% (p < 0.001).
The combination of OPRD1 (Chr1:29,139,121) hypomethylation and high levels of OPRD1 protein is a potential biomarker for gout diagnosis.
本研究旨在利用甲基化微阵列分析鉴定中国汉族男性痛风的诊断生物标志物。
我们使用甲基化微阵列筛选痛风差异甲基化基因（DMGs），并通过基因本体论（GO）分析和京都基因与基因组百科全书（KEGG）途径分析分析 DMGs 的功能。我们通过焦磷酸测序验证基因甲基化水平，通过酶联免疫吸附测定（ELISA）验证蛋白水平。统计分析采用 SPSS。所有分析均认为双侧 p 值 <0.05 具有统计学意义。
我们鉴定出 20426 个显著差异甲基化位点（5719 个高甲基化位点和 14707 个低甲基化位点）。生物信息学分析表明，DMGs 主要涉及 43 种生物学功能、13 种细胞成分、18 种分子功能和 35 种 KEGG 途径。我们选择阿片受体 delta 1（OPRD1）验证 50 例痛风患者和 50 例对照之间的甲基化水平。痛风组 OPRD1（Chr1:29139121）的甲基化水平显著降低（p<0.05），而痛风组 OPRD1 蛋白水平显著升高（p<0.05）。此外，OPRD1（Chr1:29139121）甲基化和 OPRD1 蛋白水平组合的 AUC 为 0.796（0.710，0.883），具有 82%的高灵敏度和 68%的特异性（p<0.001）。
OPRD1（Chr1:29139121）低甲基化和 OPRD1 蛋白水平升高的组合是痛风诊断的潜在生物标志物。
