METHODS: Our study enrolled 46 SLE patients with or without kidney injury and 23 healthy controls from 2019 to 2022. CD4+ T cells were sorted for DNA methylation genotyping and RNA-seq. Through bioinformatics analysis, we identified the significant differentially methylated CpG positions (DMPs) only in the LN group and validated them by Bisulfite PCR. Integration analysis was used to screen for differentially methylated and expressed genes that might be involved in the progression of LN, and the results were analyzed via cell experiments and flow cytometry.
RESULTS: We identified 243 hypomethylated sites and 778 hypermethylated sites only in the LN cohort. Three of these DMPs, cg08332381, cg03297029, and cg16797344, were validated by Bisulfite PCR and could be potential biomarkers for LN. Integrated analysis revealed that the expression of BCL2L14 and IFI27 was regulated by DNA methylation, which was validated by azacytidine (5-aza) treatment. The overexpression of BCL2L14 in CD4+ T cells might induce renal fibrosis and inflammation by regulating the differentiation and function of Tfh cells.
CONCLUSIONS: Our study identified novel aberrant DMPs in CD4+ T cells only in LN patients and DNA methylation-regulated genes that could be potential LN biomarkers. BCL2L14 is likely involved in the progression of LN and might be a treatment target.
方法:我们的研究纳入了2019年至2022年46例有或没有肾损伤的SLE患者和23例健康对照。对CD4+T细胞进行DNA甲基化基因分型和RNA-seq分选。通过生物信息学分析,我们仅在LN组中确定了显着的差异甲基化CpG位置(DMPs),并通过亚硫酸氢盐PCR进行了验证。整合分析用于筛选可能参与LN进展的差异甲基化和表达基因。并通过细胞实验和流式细胞仪分析结果。
结果:我们仅在LN队列中确定了243个低甲基化位点和778个高甲基化位点。其中三个民主党议员,cg08332381,cg03297029和cg16797344通过亚硫酸氢盐PCR验证,可能是LN的潜在生物标志物。综合分析发现BCL2L14和IFI27的表达受DNA甲基化调控,通过氮杂胞苷(5-aza)治疗验证。BCL2L14在CD4+T细胞中的过表达可能通过调节Tfh细胞的分化和功能而导致肾脏纤维化和炎症反应。
结论:我们的研究仅在LN患者中确定了CD4+T细胞中的新型异常DMPs和可能是潜在LN生物标志物的DNA甲基化调节基因。BCL2L14可能参与LN的进展,可能是治疗靶标。