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Inhibition of microRNA-24 increases liver fibrosis by enhanced menin expression in Mdr2−/− mice

  • Chad Hall
    Affiliations
    Department of Surgery, Baylor Scott & White Health and Texas A&M University Health Science Center, Temple, Texas
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  • Laurent Ehrlich
    Affiliations
    Department of Surgery, Baylor Scott & White Health and Texas A&M University Health Science Center, Temple, Texas

    Department of Medicine, Baylor Scott & White Health and Texas A&M University Health Science Center, Temple, Texas
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  • Fanyin Meng
    Affiliations
    Baylor Scott & White Digestive Disease Research Center, Baylor Scott & White, Temple, Texas
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  • Author Footnotes
    1 Present address: Program for Autoimmune Liver Diseases, International Center for Digestive Health, Department of Medicine and Surgery, University of Milan-Bicocca, Milan, Italy.
    Pietro Invernizzi
    Footnotes
    1 Present address: Program for Autoimmune Liver Diseases, International Center for Digestive Health, Department of Medicine and Surgery, University of Milan-Bicocca, Milan, Italy.
    Affiliations
    Center for Autoimmune Liver Diseases, Humanitas Clinical and Research Center, Rozzano, Italy
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  • Author Footnotes
    1 Present address: Program for Autoimmune Liver Diseases, International Center for Digestive Health, Department of Medicine and Surgery, University of Milan-Bicocca, Milan, Italy.
    Francesca Bernuzzi
    Footnotes
    1 Present address: Program for Autoimmune Liver Diseases, International Center for Digestive Health, Department of Medicine and Surgery, University of Milan-Bicocca, Milan, Italy.
    Affiliations
    Center for Autoimmune Liver Diseases, Humanitas Clinical and Research Center, Rozzano, Italy
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  • Terry C. Lairmore
    Affiliations
    Department of Surgery, Baylor Scott & White Health and Texas A&M University Health Science Center, Temple, Texas
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  • Gianfranco Alpini
    Affiliations
    Department of Medicine, Baylor Scott & White Health and Texas A&M University Health Science Center, Temple, Texas

    Baylor Scott & White Digestive Disease Research Center, Baylor Scott & White, Temple, Texas

    Research, Central Texas Veterans Health Care System, Temple, Texas
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  • Shannon Glaser
    Correspondence
    Corresponding author. Scott & White Digestive Diseases Research Center, Central Texas Veterans University Health Care System, Texas A & M Health Science Center, Olin E. Teague Medical Center, 1901 South 1st Street, Bldg. 205, Temple, TX 76504. Tel.: +1 254 743-1044; fax: +1 254 743-0378.
    Affiliations
    Department of Medicine, Baylor Scott & White Health and Texas A&M University Health Science Center, Temple, Texas

    Baylor Scott & White Digestive Disease Research Center, Baylor Scott & White, Temple, Texas

    Research, Central Texas Veterans Health Care System, Temple, Texas
    Search for articles by this author
  • Author Footnotes
    1 Present address: Program for Autoimmune Liver Diseases, International Center for Digestive Health, Department of Medicine and Surgery, University of Milan-Bicocca, Milan, Italy.

      Abstract

      Background

      Liver transplantation remains the primary treatment for primary sclerosing cholangitis (PSC). Mdr2−/− mice provide a reliable in vivo model of PSC and develop characteristic biliary inflammation and fibrosis. We tested the hypothesis that the tumor suppressor protein menin is implicated in the progression of liver fibrosis and that menin expression can be regulated in the liver via microRNA-24 (miR-24).

      Materials and methods

      Menin expression was measured in human PSC and Mdr2−/− mice. Twelve-week-old FVB/NJ wild-type (WT) and Mdr2−/− mice were treated with miR-24 Vivo-Morpholino to knockdown miR-24 expression levels. Liver fibrosis was evaluated by Sirius Red staining and quantitative polymerase chain reaction (qPCR) for genes associated with liver fibrosis, such as fibronectin 1, collagen type 1 alpha 1, transforming growth factor-β1 (TGF-β1), and α-smooth muscle actin. Studies were also performed in vitro using immortalized murine cholangiocyte lines treated with miR-24 hairpin inhibitor and mimic.

      Results

      Menin gene expression was increased in Mdr2−/− mice and late-stage human PSC samples. Treatment of FVB/NJ WT and Mdr2−/− mice with miR-24 Vivo-Morpholino increased menin expression, which correlated with increased expression of fibrosis genes. In vitro, inhibition of miR-24 also significantly increased the expression of fibrosis genes.

      Conclusions

      Inhibition of miR-24 increases menin and TGF-β1 expression, subsequently increasing hepatic fibrosis in FVB/NJ WT and Mdr2−/− mice. Modulation of the menin/miR-24 axis may provide novel targeted therapies to slow the progression of hepatic fibrosis into cirrhosis in PSC patients by altering TGF-β1 expression.

      Keywords

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