miR-20a suppresses Treg differentiation by targeting Map3k9 in experimental autoimmune encephalomyelitis.
miR-20a suppresses Treg differentiation by targeting Map3k9 in experimental autoimmune encephalomyelitis.
复制标题
实验性自身免疫性脑脊髓炎中 miR-20a 通过靶向 Map3k9 抑制 Treg 分化
DOI：
10.1186/s12967-021-02893-4
复制
发表时间：
2021-05-26
影响因子：
7.4
通讯作者：
Guan Y
中科院分区：
文献类型：
--
作者：
Wang Y;Xie C;Song Y;Xiang W;Peng J;Han L;Ding J;Guan Y
实验性自身免疫性脑脊髓炎（EAE）是中枢神经系统（CNS）炎性脱髓鞘疾病的一种模型，这是一组以炎性浸润、脱髓鞘和轴突损伤为特征的自身免疫性疾病。miR - 20a在中枢神经系统炎性脱髓鞘疾病患者中表达失调；然而，miR - 20a的功能仍不清楚。在本研究中，我们旨在探究miR - 20a在EAE中的作用。
通过实时定量聚合酶链反应（qRT - PCR）检测EAE小鼠以及抗髓鞘少突胶质细胞糖蛋白（MOG）抗体相关脱髓鞘疾病患者中miR - 20a的表达。对EAE小鼠的CD4⁺T细胞进行分选、刺激，并在miR - 20a敲低的情况下进行极化。通过流式细胞术分析CD4⁺T细胞的活化和分化。通过qRT - PCR和蛋白质印迹实验检测靶基因Map3k9的表达。通过荧光素酶试验检测miR - 20a与Map3k9的3’非翻译区（3’ UTR）的结合。通过向EAE小鼠静脉注射miR - 20a拮抗剂来探索miR - 20a作为减轻EAE严重程度的治疗靶点的可行性。
miR - 20a在EAE小鼠的脾细胞和淋巴结细胞、CD4⁺T细胞以及脊髓中表达上调。此外，miR - 20a敲低不影响抗原特异性CD4⁺T细胞的活化，但促进其分化为调节性T细胞（Treg细胞）。Map3k9被预测为miR - 20a的一个靶基因。Map3k9和miR - 20a的表达呈负相关，并且miR - 20a敲低会增加Map3k9的表达。此外，miR - 20a与Map3k9的3’ UTR结合，同时敲低miR - 20a和Map3k9抵消了单独敲低miR - 20a时观察到的Tregs分化增强的现象。再者，向EAE小鼠注射miR - 20a拮抗剂降低了疾病的严重程度，并增加了外周免疫器官中Treg细胞的比例。
miR - 20a通过降低Map3k9的表达抑制EAE中抗原特异性CD4⁺T细胞向Tregs的分化。miR - 20a拮抗剂可缓解EAE，这为EAE和中枢神经系统炎性脱髓鞘疾病提供了一种新的治疗方法。
网络版包含补充材料，可在10.1186/s12967 - 2021 - 02893 - 4获取。
Background
Experimental autoimmune encephalomyelitis (EAE) is a model for inflammatory demyelinating diseases of the central nervous system (CNS), a group of autoimmune diseases characterized by inflammatory infiltration, demyelination, and axonal damage. miR-20a is dysregulated in patients with CNS inflammatory demyelinating diseases; however, the function of miR-20a remains unclear. In this study, we intended to explore the role of miR-20a in EAE.
Methods
The expression of miR-20a was detected by quantitative real-time PCR (qRT-PCR) in EAE mice and patients with MOG antibody-associated demyelinating diseases. CD4+ T cells of EAE mice were sorted, stimulated, and polarized with miR-20a knockdown. Activation and differentiation of CD4+ T cells were analyzed by flow cytometry. The expression of target gene Map3k9 was detected by qRT-PCR and western blot experiments. The binding of miR-20a to the 3’ UTR of Map3k9 was tested by luciferase assays. The feasibility of miR-20a as a therapeutic target to alleviate the severity of EAE was explored by intravenous administration of miR-20a antagomirs to EAE mice.
Results
miR-20a was upregulated in splenocytes and lymph node cells, CD4+ T cells, and spinal cords of EAE mice. Moreover, miR-20a knockdown did not influence the activation of antigen-specific CD4+ T cells but promoted their differentiation into Treg cells. Map3k9 was predicted to be a target gene of miR-20a. The expressions of Map3k9 and miR-20a were negatively correlated, and miR-20a knockdown increased the expression of Map3k9. In addition, miR-20a binded to the 3’ UTR of Map3k9, and simultaneous knockdown of miR-20a and Map3k9 counteracted the enhanced differentiation of Tregs observed when miR-20a was knocked down alone. Furthermore, injection of miR-20a antagomirs to EAE mice reduced the severity of the disease and increased the proportion of Treg cells in peripheral immune organs.
Conclusions
miR-20a suppresses the differentiation of antigen-specific CD4+ T cells into Tregs in EAE by decreasing the expression of Map3k9. miR-20a antagomirs alleviate EAE, suggesting a new therapy for EAE and CNS inflammatory demyelinating diseases.
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