Identification of microRNA-31 as a novel regulator contributing to impaired interleukin-2 production in T cells from patients with systemic lupus erythematosus

Authors

  • Wei Fan,

    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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    • Drs. Fan, Liang, and Tang contributed equally to this work.

  • Dong Liang,

    Corresponding author
    1. Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio
    • Division of Rheumatology, Center for Autoimmune Genomics and Etiology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, Ohio 45229
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    • Drs. Fan, Liang, and Tang contributed equally to this work.

  • Yuanjia Tang,

    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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    • Drs. Fan, Liang, and Tang contributed equally to this work.

  • Bo Qu,

    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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  • Huijuan Cui,

    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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  • Xiaobing Luo,

    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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  • Xinfang Huang,

    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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  • Shunle Chen,

    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
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  • Brandon W. Higgs,

    1. MedImmune, Gaithersburg, Maryland
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    • Drs. Higgs, Jallal, and Yao own stock or stock options in AstraZeneca.

  • Bahija Jallal,

    1. MedImmune, Gaithersburg, Maryland
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    • Drs. Higgs, Jallal, and Yao own stock or stock options in AstraZeneca.

  • Yihong Yao,

    1. MedImmune, Gaithersburg, Maryland
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    • Drs. Higgs, Jallal, and Yao own stock or stock options in AstraZeneca.

  • John B. Harley,

    1. Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio
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  • Nan Shen

    Corresponding author
    1. Joint Molecular Rheumatology Laboratory of the Institute of Health Sciences and Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China
    2. Division of Rheumatology and the Center for Autoimmune Genomics and Etiology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio
    • Department of Rheumatology, Shanghai Renji Hospital, Shanghai JiaoTong University School of Medicine, 145 Shan Dong Road (Central), Shanghai 200001, China
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Abstract

Objective

MicroRNAs (miRNAs) function to fine-tune the control of immune cell signaling. It is well established that there are abnormalities in the interleukin-2 (IL-2)–related signaling pathways in systemic lupus erythematosus (SLE). The miR-31 microRNA has been found to be markedly underexpressed in patients with SLE, and thus the present study was undertaken to investigate the role of miR-31 in IL-2 defects in lupus T cells.

Methods

Expression levels of miR-31 were quantitated using TaqMan miRNA assays. Transfection and stimulation of cultured cells followed by TaqMan quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, and reporter gene assays were conducted to determine the biologic function of miR-31. NF-AT nuclear translocation and expression were quantitatively measured using an ImageStream cytometer. Bioinformatics analysis, small interfering RNA (siRNA) knockdown, and Western blotting were performed to validate miR-31 targets and effects.

Results

The expression of miR-31 was significantly decreased in lupus T cells, and this was positively correlated with the expression of IL-2. Overexpression of miR-31 in T cells increased the production of IL-2 by altering NF-AT nuclear expression and IL2 promoter activity, while knockdown of endogenous miR-31 reduced IL-2 production. RhoA expression was directly repressed by miR-31 in T cells. Of note, siRNA-mediated knockdown of RhoA enhanced IL2 promoter activity and, consequently, up-regulated IL-2 production. RhoA expression was consistently up-regulated and negatively correlated with the levels of miR-31 in lupus T cells. Manipulation of miR-31 expression in lupus T cells restored the expression of IL-2 at both the messenger RNA and protein levels.

Conclusion

MicroRNA-31 is a novel enhancer of IL-2 production during T cell activation. Dysregulation of miR-31 and its target, RhoA, could be a novel molecular mechanism underlying the IL-2 deficiency in patients with SLE.

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