目的:我们通过检测系统性红斑狼疮(SLE)患者的lncRNA表达谱,初步探索差异表达的长链非编码RNA(lncRNA)与调节性T(Treg)细胞数量之间的联系,然后分析Treg相关lncRNAs与SLE患者临床特征的相关性,预测lncRNAs调控Treg细胞分化和发育的机制,为SLE的治疗提供了新的思路。
方法:收集9例活动性SLE患者的外周血,提取单核细胞(PBMC);通过全转录组测序分析PBMC的lncRNA表达谱。9名健康人作为对照筛选差异表达的lncRNAs,分析lncRNAs与Treg细胞数的相关性。采用Pearson检验分析lncRNAs与Treg细胞数的相关性,以及Treg相关lncRNA与SLEDAI评分之间的相关性,ESR,SLE患者的C3和C4。用miRcode和Targetscan数据库和共表达网络预测Treg相关lncRNAs的靶基因。
结果:与健康对照相比,SLE患者中有240个差异表达的lncRNAs,包括134个高表达的lncRNAs(p<0.05)和106个低表达的lncRNAs(p<0.05)。ANKRD44-AS1的表达(r=0.7417,p=0.0222),LINC00200(r=0.6960,p=0.0373),AP001363.2(r=0.7766,p=0.0138),LINC02824(r=0.7893,p=0.0114)与Treg细胞数呈正相关,和AP000640.1的表达式(r=-0.7225,p=0.0279),AC124248.1(r=-0.7653,p=0.0163),LINC00482(r=-0.8317,p=0.0054),MIR503HG(r=-0.7617,p<0.05)与Treg细胞数呈负相关。在这些Treg相关的lncRNAs中,LINC00482(r=-0.7348,p<0.05)和HGMIR503(r=-0.7617,p<0.05)的表达与C3呈负相关。与Treg细胞相关的LINC00200、ANKRD44-AS1和AP000640.1调节信号转导和转录激活因子5(STAT5)的表达,磷脂酶D1(PLD1),仅同源结构域蛋白X(HOPX),和runt相关转录因子3(RUNX3)通过miRNA的竞争性结合或反式调节机制,从而调控Treg细胞的分化和发展。
结论:SLE患者的lncRNA表达谱发生了变化,差异表达的lncRNAs与SLE中Treg细胞的数量和功能异常相关,和Treg相关的lncRNAs与SLE疾病活动相关,这可能会影响STAT5,PLD1,HOPX的表达,RUNX3和调节Treg细胞功能,并通过竞争性结合miRNA或反式调节机制参与SLE的发病和进展。要点•系统性红斑狼疮(SLE)是涉及多个器官和系统的自身免疫性疾病。lncRNAs可能通过调节基因表达影响Treg细胞功能,这可能是SLE的重要发病机制。•这项研究,以SLE为例,初步分析lncRNA与SLE患者Treg细胞的相关性,分析Treg相关lncRNA与SLE临床特征的相关性,并推测lncRNA可以通过与miRNA竞争结合或反式调节机制调控Treg细胞的分化和发育。•有可能针对SLE的表观遗传疗法。
OBJECTIVE: We initially explored the link between the differentially expressed long non-coding RNAs (lncRNAs) and the number of regulatory T (Treg) cells by detecting the lncRNA expression profiles in patients with
systemic lupus erythematosus (SLE), then analyzed the correlation between Treg-related lncRNAs and the clinical features of SLE patients, predicting the mechanism by which lncRNAs regulate the differentiation and development of Treg cells, and provided new ideas for the treatment of SLE.
METHODS: Peripheral blood of 9 active SLE patients were collected and mononuclear cells (PBMCs) were extracted; the lncRNA expression profiles of PBMCs were analyzed by whole transcriptome sequencing. Nine healthy people were used as controls to screen the differentially expressed lncRNAs, to analyze the correlation between lncRNAs and Treg cell number. Pearson test was used to analyze the correlation between lncRNAs and the number of Treg cell, and the correlation between Treg-associated lncRNA and SLEDAI score, ESR, C3, and C4 in SLE patients. The targeted genes of Treg-associated lncRNAs were predicted with miRcode and Targetscan databases and coexpression network.
RESULTS: There were 240 differentially expressed lncRNAs in SLE patients compared with healthy controls, including 134 highly expressed lncRNAs (p < 0.05) and 106 lowly expressed lncRNAs (p < 0.05). The expression of ANKRD44-AS1 (r = 0.7417, p = 0.0222), LINC00200 (r = 0.6960, p = 0.0373), AP001363.2 (r = 0.7766, p = 0.0138), and LINC02824 (r = 0.7893, p = 0.0114) were positively correlated with the number of Treg cell, and the expression of AP000640.1 (r = - 0.7225, p = 0.0279), AC124248.1 (r = - 0.7653, p = 0.0163), LINC00482 (r = - 0.8317, p = 0.0054), and MIR503HG (r = - 0.7617, p < 0.05) were negatively correlated with the number of Treg cell. Among these Treg-associated lncRNAs, the expression of LINC00482 (r = - 0.7348, p < 0.05) and MIR503 HG (r = - 0.7617, p < 0.05) were negatively correlated with C3. LINC00200, ANKRD44 - AS1, and AP000640.1 related to Treg cells regulate the expression of signal transducer and activator of transcription 5 (STAT5), phospholipase D1 (PLD1), homeodomain-only protein X (HOPX), and runt-related transcription factor 3 (RUNX3) through competitive binding of miRNA or trans-regulatory mechanism, thereby regulating the differentiation and development of Treg cell.
CONCLUSIONS: The lncRNA expression profiles were changed in SLE patients, the differentially expressed lncRNAs were associated with abnormal number and function of Treg cells in SLE, and Treg-associated lncRNAs were associated with SLE-disease activity, which may affect the expression of STAT5, PLD1, HOPX, RUNX3 and regulate Treg cell function and participate in the pathogenesis and progression of SLE by competitively binding to miRNAs or trans-regulatory mechanism. Key points •
Systemic lupus erythematosus (SLE) is an autoimmune disease involving multiple organs and systems. lncRNAs may affect Treg cells function by regulating genes expression, which may be an important pathogenesis of SLE. • This
study, taking SLE as an example, preliminarily analyzed the correlation between lncRNA and Treg cells in SLE patients, analyzed the correlation between Treg-related lncRNA and the clinical characteristics of SLE, and speculated that lncRNA could regulate the differentiation and development of Treg cells through competitive combination with miRNA or trans-regulatory mechanisms. • It is possible to target epigenetic therapy for SLE.