title: miR-425 regulates inflammatory cytokine production in CD4+T cells via N-Ras upregulation in primary biliary cholangitis.
abstract: Primary biliary cholangitis (PBC) is an autoimmune liver disease of unknown pathogenesis. Consequently, therapeutic targets for PBC have yet to be identified. CD4+T cells play a pivotal role in immunological dysfunction observed in PBC, and therefore, microRNA (miRNA) and mRNA expression were analysed in CD4+T cells, to investigate PBC pathogenesis and identify novel therapeutic targets.
  
  
  
  Integral miRNA and mRNA analysis of 14 PBC patients and ten healthy controls was carried out using microarray and quantitative real-time polymerase chain reaction (qRT-PCR), with gene set enrichment analysis. The functional analyses of miRNA were then assessed using reporter and miRNA-overexpression assays.
  
  
  
  The integral analysis of miRNA and mRNA identified four significantly downregulated miRNAs (miR-181a, -181b, -374b, and -425) related to the T cell receptor (TCR) signalling pathway in CD4+T cells of PBC. N-Ras, a regulator of the TCR signalling pathway, was found to be targeted by all four identified miRNAs. In addition, in vitro assays confirmed that decreased miR-425 strongly induced inflammatory cytokines (interleukin [IL]-2 and interferon [IFN]-γ) via N-Ras upregulation in the TCR signalling pathway.
  
  
  
  The decreased expression of four miRNAs that dysregulate TCR signalling in PBC CD4+T cells was identified. miR-425 was demonstrated as an inflammatory regulator of PBC via N-Ras upregulation. Therefore, the restoration of decreased miR-425 or the suppression of N-Ras may be a promising immunotherapeutic strategy against PBC.
  
  
  
  Primary biliary cholangitis (PBC) is an autoimmune liver disease, but the causes are unknown. MicroRNAs are molecules known to regulate biological signals. In this study, four microRNAs were identified as being decreased in PBC patients, leading to activation of T cell receptor signalling pathways, involved in inflammation. One particular target, N-Ras, could be an attractive and novel immunotherapeutic option for PBC.
  
  
  
  Microarray data are deposited in GEO (GEO accession: GSE93172).
authors.affiliation: Division of Advanced Genome Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan; Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Ryo
authors.initials: R
authors.lastname: Nakagawa
authors.affiliation: Division of Advanced Genome Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Muroyama
authors.affiliation: Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Chisato
authors.initials: C
authors.lastname: Saeki
authors.affiliation: Division of Advanced Genome Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Kaku
authors.initials: K
authors.lastname: Goto
authors.affiliation: Division of Advanced Genome Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Kaise
authors.affiliation: Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Koike
authors.affiliation: Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Masanori
authors.initials: M
authors.lastname: Nakano
authors.affiliation: Division of Advanced Genome Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Matsubara
authors.affiliation: Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Keiko
authors.initials: K
authors.lastname: Takano
authors.affiliation: Division of Advanced Genome Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Ito
authors.affiliation: Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Saruta
authors.affiliation: Division of Advanced Genome Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan. Electronic address: kato-2im@ims.u-tokyo.ac.jp.
authors.forename: Naoya
authors.initials: N
authors.lastname: Kato
authors.affiliation: Department of Gastroenterology and Hepatology, The Jikei University School of Medicine, Tokyo, Japan; Sanno Medical Center, Sanno Hospital, International University of Health and Welfare, Tokyo, Japan.
authors.forename: Mikio
authors.initials: M
authors.lastname: Zeniya
mesh_terms: D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D016022:Case-Control Studies; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D005204:Farnesol / Q000031:analogs & derivatives / Q000494:pharmacology; D020869:Gene Expression Profiling; D011905:Genes, ras; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics; D019169:Jurkat Cells; D008105:Liver Cirrhosis, Biliary / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D012459:Salicylates / Q000494:pharmacology; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D015854:Up-Regulation
chemical_list: D016207:Cytokines; C573415:IFNG protein, human; C508594:IL2 protein, human; D018836:Inflammation Mediators; D007376:Interleukin-2; C555677:MIRN425 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D012459:Salicylates; C093323:farnesylthiosalicylic acid; D005204:Farnesol; D007371:Interferon-gamma
keywords: Autoimmune liver disease; Cholestatic liver disease; Helper T cell; Small noncoding RNA; T cell receptor signalling pathway