A Novel Long Noncoding RNA lincRNA00892 Activates CD4(+) T Cells in Systemic Lupus Erythematosus by Regulating CD40L.
A Novel Long Noncoding RNA lincRNA00892 Activates CD4(+) T Cells in Systemic Lupus Erythematosus by Regulating CD40L.
复制标题
新型长非编码RNA lincRNA00892通过调节CD40L激活系统性红斑狼疮中的CD4( ) T细胞
DOI：
10.3389/fphar.2021.733902
复制
发表时间：
2021
影响因子：
5.6
通讯作者：
Xu J
中科院分区：
文献类型：
--
作者：
Liu X;Lin J;Wu H;Wang Y;Xie L;Wu J;Qin H;Xu J
目的：系统性红斑狼疮(SLE)患者外周血中CD4+T细胞功能障碍的机制尚不完全清楚。越来越多的证据表明，长非编码RNA(Long Non Coding RNAs，LncRNAs)可以调节免疫反应，参与某些自身免疫性疾病的发生，但对LncRNA在SLE患者CD4+T细胞中的表达和功能却知之甚少。本研究旨在检测狼疮患者CD4+T细胞中lncRNAs的表达谱，探讨其在SLE发病机制中的作用机制。方法：应用基因芯片技术检测SLE患者和正常人外周血中CD4+T细胞中lncRNAs和mRNAs的表达谱。选择LincRNA00892和CD40L进行实时定量聚合酶链式反应(qRT-PCR)验证。利用共表达网络预测lincRNA00892的潜在靶基因。经慢病毒感染后，lincRNA00892在正常的CD4+T细胞中高表达。QRT-PCR检测lincRNA00892的表达。采用定量逆转录聚合酶链式反应、Western blotting和流式细胞仪检测CD40L的表达。用流式细胞仪检测CD69和CD23的表达。采用酶联免疫吸附试验(EL ISA)检测免疫球蛋白G的分泌。以lincRNA00892为靶向的蛋白质通过RNA下拉和随后的质谱仪(MS)进行检测。用RNA免疫沉淀(RIP)法检测异质性核糖核蛋白K(HnRNP K)与lincRNA00892或CD40L的相互作用。结果：SLE患者外周血中存在1887个lncRNAs和3375mRNAs的异常表达。定量逆转录聚合酶链式反应证实SLE患者外周血中存在LincRNA00892和CD40L的上调。LncRNA-mRNA共表达网络分析表明，CD40L是lincRNA00892的潜在靶点。过表达lincRNA00892可增强CD4+T细胞CD40L蛋白水平，而对CD40L mRNA水平影响不大。此外，lincRNA00892还能诱导CD4+T细胞活化。此外，lincRNA00892以一种依赖于CD4+T细胞的方式导致B细胞的激活和随后的免疫球蛋白G的分泌。最后发现，hnRNP K是lincRNA00892下调的蛋白质中的一种，hnRNP K可以直接与lincRNA00892或CD40L结合。结论：SLE患者CD4+T细胞中的lncRNA表达谱发生了改变。LincRNA00892可能通过靶向hnRNP K，上调CD40L的表达来激活CD4+T和B细胞，从而参与SLE的发病。这为进一步研究SLE的发病机制提供了一个潜在的靶点。
Objective: The mechanism of CD4+ T-cell dysfunction in systemic lupus erythematosus (SLE) has not been fully understood. Increasing evidence show that long noncoding RNAs (lncRNAs) can regulate immune responses and take part in some autoimmune diseases, while little is known about the lncRNA expression and function in CD4+ T of SLE. Here, we aimed to detect the expression profile of lncRNAs in lupus CD4+ T cells and explore the mechanism that how lincRNA00892 in CD4+ T cells is involved in the pathogenesis of SLE. Methods: The expression profiles of lncRNAs and mRNAs in CD4+ T cells from SLE patients and healthy controls were detected by microarray. LincRNA00892 and CD40L were chosen for validation by quantitative real-time PCR (qRT-PCR). Coexpression network was conducted to predict the potential target genes of lincRNA00892. Then lincRNA00892 was overexpressed in normal CD4+ T cells via lentivirus transfection. The expression of lincRNA00892 was detected by qRT-PCR. The expression of CD40L was detected by qRT-PCR, western blotting, and flow cytometry, respectively. The expression of CD69 and CD23 was measured by flow cytometry. The secretion of IgG was determined by enzyme-linked immunosorbent assay (ELISA). The proteins targeted by lincRNA00892 were measured by RNA pulldown and subsequent mass spectrometry (MS). The interaction between heterogeneous nuclear ribonucleoprotein K (hnRNP K) and lincRNA00892 or CD40L was detected by RNA immunoprecipitation (RIP) assay. Results: A total of 1887 lncRNAs and 3375 mRNAs were found to be aberrantly expressed in CD4+ T cells of SLE patients compared to healthy controls. LincRNA00892 and CD40L were confirmed to be upregulated in CD4+ T cells of SLE patients by qRT-PCR. The lncRNA–mRNA coexpression network analysis indicated that CD40L was a potential target of lincRNA00892. Overexpression of lincRNA00892 enhanced CD40L protein levels while exerting little influence on CD40L mRNA levels in CD4+ T cells. In addition, lincRNA00892 could induce the activation of CD4+ T cells. Furthermore, lincRNA00892 led to the activation of B cells and subsequent secretion of IgG in a CD4+ T-cell–dependent manner. Finally, hnRNP K was found to be among the proteins pulled down by lincRNA00892, and hnRNP K could bind to lincRNA00892 or CD40L directly. Conclusion: Our results showed that the lncRNA expression profile was altered in CD4+ T cells of SLE. LincRNA00892 possibly contributed to the pathogenesis of SLE by targeting hnRNP K and subsequently upregulating CD40L expression to activate CD4+ T and B cells. These provided us a potential target for further mechanistic studies of SLE pathogenesis.
登录
查看更多内容
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
4.8
作者：
Hamilton, B. JoNell;Wang, Xiao-Wei;Rigby, William F. C.
通讯作者：
Rigby, William F. C.
影响因子：
8.6
作者：
Liao, J.;Liang, G.;Lu, Q.
通讯作者：
Lu, Q.
DOI：
10.1084/jem.175.4.1091
发表时间：
1992-04-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Lederman S;Yellin MJ;Krichevsky A;Belko J;Lee JJ;Chess L
通讯作者：
Chess L
DOI：
10.1126/science.1237973
发表时间：
2013-08-16
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Engreitz JM;Pandya-Jones A;McDonel P;Shishkin A;Sirokman K;Surka C;Kadri S;Xing J;Goren A;Lander ES;Plath K;Guttman M
通讯作者：
Guttman M