Identification of Renal Long Non-coding RNA RP11-2B6.2 as a Positive Regulator of Type I Interferon Signaling Pathway in Lupus Nephritis
Identification of Renal Long Non-coding RNA RP11-2B6.2 as a Positive Regulator of Type I Interferon Signaling Pathway in Lupus Nephritis
复制标题
鉴定肾长非编码 RNA RP11-2B6.2 作为狼疮性肾炎 I 型干扰素信号通路的正调节因子
DOI：
10.3389/fimmu.2019.00975
复制
发表时间：
2019-05-03
影响因子：
7.3
通讯作者：
Shen, Nan
中科院分区：
文献类型：
--
作者：
Liao, Zhuojun;Ye, Zhizhong;Shen, Nan
目的：狼疮性肾炎（LN）是系统性红斑狼疮（SLE）最严重的并发症之一。I型干扰素（IFN-1）与LN的发病有关。长链非编码RNA（lncRNA）与SLE的发病机制有关，但lncRNA在LN中的作用仍知之甚少。在此，我们鉴定并研究了LN相关的lncRNA RP 11 -2B6.2在调节IFN-1信号通路中的功能。方法：应用RNA测序技术分析LN患者和正常对照肾组织中lncRNA的表达。使用反义寡核苷酸和CRISPRi系统或过表达质粒和CRISPRa系统进行功能丧失或获得实验。采用原位杂交、成像流式细胞术、双荧光素酶报告基因分析和ATAC测序等方法研究lncRNA RP 11 -2B6.2的功能。采用RT-qPCR、ELISA和Western blotting检测特异基因的RNA和蛋白水平。结果：LN患者肾活检组织中lncRNA RP 11 -2B6.2水平升高，与疾病活动度和IFN评分呈正相关。在肾细胞中敲低lncRNA RP 11 -2B6.2抑制IFN刺激基因（ISG）的表达，而过表达lncRNA RP 11 -2B6.2增强ISG表达。敲低LncRNA RP 11 -2B6.2可抑制IFN-1通路中JAK 1、TYK 2和STAT 1的磷酸化，同时促进染色质可及性和SOCS 1的转录。结论：LN患者肾脏中lncRNA表达异常。LncRNA RP 11 -2B6.2通过表观遗传学抑制SOCS 1而成为IFN-I通路的新型正调控因子，为减轻LN中过度激活的IFN-I信号通路提供了新的治疗靶点。
Objective: Lupus nephritis (LN) is one of the most serious complications of systemic lupus erythematosus (SLE). Type I interferon (IFN-I) is associated with the pathogenesis of LN. Long non-coding RNAs (lncRNAs) have been implicated in the pathogenesis of SLE, however, the roles of lncRNAs in LN are still poorly understood. Here, we identified and investigated the function of LN-associated lncRNA RP11-2B6.2 in regulating IFN-I signaling pathway. Methods: RNA sequencing was used to analyze the expression of lncRNAs in kidney biopsies from LN patients and controls. Antisense oligonucleotides and CRISPRi system or overexpression plasmids and CRISPRa system were used to perform loss or gain of function experiments. In situ hybridization, imaging flow cytometry, dual-luciferase reporter assay, and ATAC sequencing were used to study the functions of lncRNA RP11-2B6.2. RT-qPCR, ELISA, and western blotting were done to detect RNA and protein levels of specific genes. Results: Elevated lncRNA RP11-2B6.2 was observed in kidney biopsies from LN patients and positively correlated with disease activity and IFN scores. Knockdown of lncRNA RP11-2B6.2 in renal cells inhibited the expression of IFN stimulated genes (ISGs), while overexpression of lncRNA RP11-2B6.2 enhanced ISG expression. Knockdown of LncRNA RP11-2B6.2 inhibited the phosphorylation of JAK1, TYK2, and STAT1 in IFN-I pathway, while promoted the chromatin accessibility and the transcription of SOCS1. Conclusion: The expression of lncRNAs is abnormal in the kidney of LN. LncRNA RP11-2B6.2 is a novel positive regulator of IFN-I pathway through epigenetic inhibition of SOCS1, which provides a new therapeutic target to alleviate over-activated IFN-I signaling in LN.