Downregulation of miR-200a-3p, Targeting CtBP2 Complex, Is Involved in the Hypoproduction of IL-2 in Systemic Lupus Erythematosus-Derived T Cells

• Published in: The Journal of immunology (1950) Vol. 198; no. 11; pp. 4268 - 4276
• Main Authors: Katsuyama, Eri, Yan, Minglu, Watanabe, Katsue Sunahori, Narazaki, Mariko, Matsushima, Syun, Yamamura, Yuriko, Hiramatsu, Sumie, Ohashi, Keiji, Watanabe, Haruki, Katsuyama, Takayuki, Zeggar, Sonia, Yoshida, Nobuya, Moulton, Vaishali R, Tsokos, George C, Sada, Ken-Ei, Wada, Jun
• Format: Journal Article
• Language: English
• Published: United States American Association of Immunologists 01.06.2017
• Subjects: Animals; Autoantibodies; Autoimmune diseases; CD4 antigen; Cell Line; Cell lines; Chronic conditions; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Down-Regulation; Homeobox; Immunological tolerance; Interleukin 2; Interleukin-2 - biosynthesis; Interleukin-2 - genetics; Interleukin-2 - immunology; Ionomycin; Lupus; Lupus Erythematosus, Systemic - immunology; Lupus Erythematosus, Systemic - metabolism; Lymphocytes; Lymphocytes T; Mice; Mice, Inbred C57BL; Mice, Inbred MRL lpr; MicroRNAs - genetics; miRNA; Organs; Pathogenesis; Phosphoproteins - genetics; Phosphoproteins - metabolism; Stimulation; Systemic lupus erythematosus; T-Lymphocytes - immunology; T-Lymphocytes - pathology; Transcriptional Activation; Zinc; Zinc Finger E-box-Binding Homeobox 1 - metabolism; Zinc finger proteins
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.1601705

Systemic lupus erythematosus (SLE) damages multiple organs by producing various autoantibodies. In this study, we report that decreased microRNA (miR)-200a-3p causes IL-2 hypoproduction through zinc finger E-box binding homeobox (ZEB)1 and C-terminal binding protein 2 (CtBP2) in a lupus-prone mouse. First, we performed RNA sequencing to identify candidate microRNAs and mRNAs involved in the pathogenesis of SLE. We found that miR-200a-3p was significantly downregulated, whereas its putative targets, ZEB2 and CtBP2, were upregulated in CD4 T cells from MRL/lpr- mice compared with C57BL/6J mice. ZEB1 and ZEB2 comprise the ZEB family and suppress various genes, including IL-2 by recruiting CtBP2. IL-2 plays a critical role in immune tolerance, and insufficient IL-2 production upon stimulation has been recognized in SLE pathogenesis. Therefore, we hypothesized that decreased miR-200a-3p causes IL-2 deficit through the ZEB1-CtBP2 and/or ZEB2-CtBP2 complex in SLE CD4 T cells. Overexpression of miR-200a-3p induced IL-2 production by downregulating ZEB1, ZEB2, and CtBP2 in EL4 cell lines. We further revealed that miR-200a-3p promotes IL-2 expression by reducing the binding of suppressive ZEB1-CtBP2 and ZEB2-CtBP2 complexes on negative regulatory element A in the IL-2 promoter in EL4 cells. Interestingly, the ZEB1-CtBP2 complex on negative regulatory element A was significantly upregulated after PMA/ionomycin stimulation in lupus CD4 T cells. Our studies have revealed a new epigenetic pathway in the control of IL-2 production in SLE whereby low levels of miR-200a-3p accumulate the binding of the ZEB1-CtBP2 complex to the IL-2 promoter and suppress IL-2 production.