Advertisement

MicroRNA-378 promotes hepatic inflammation and fibrosis via modulation of the NF-κB-TNFα pathway

  • Author Footnotes
    † Tianpeng Zhang and Junjie Hu contributed equally to this work.
    Tianpeng Zhang
    Footnotes
    † Tianpeng Zhang and Junjie Hu contributed equally to this work.
    Affiliations
    Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA
    Search for articles by this author
  • Author Footnotes
    † Tianpeng Zhang and Junjie Hu contributed equally to this work.
    Junjie Hu
    Footnotes
    † Tianpeng Zhang and Junjie Hu contributed equally to this work.
    Affiliations
    Key Laboratory of Chinese Medicine Resource and Compound Prescription, Ministry of Education, Hubei University of Chinese Medicine, Wuhan, Hubei Province 430065, China
    Search for articles by this author
  • Xiaomei Wang
    Affiliations
    Institute for Translational Medicine, Jilin University, Changchun, Jilin Province 130021, China
    Search for articles by this author
  • Xiaoling Zhao
    Affiliations
    McLab, South San Francisco, CA 94080, USA
    Search for articles by this author
  • Zhuoyu Li
    Affiliations
    Institute of Biotechnology, Shanxi University, Taiyuan, Shanxi Province 030006, China
    Search for articles by this author
  • Junqi Niu
    Affiliations
    Key Laboratory of Chinese Medicine Resource and Compound Prescription, Ministry of Education, Hubei University of Chinese Medicine, Wuhan, Hubei Province 430065, China
    Search for articles by this author
  • Clifford J. Steer
    Affiliations
    Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA

    Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA
    Search for articles by this author
  • Guohua Zheng
    Correspondence
    Corresponding authors. Address: Division of Gastroenterology, Hepatology and Nutrition, University of Minnesota, 516 Delaware Street SE, Minneapolis, MN 55455, USA (G. Song), or Key Laboratory of Chinese Medicine Resource and Compound Prescription, Ministry of Education, Hubei University of Chinese Medicine, 1 West Huangjiahu Road, Wuhan, Hubei Province 430065, China (G. Zheng).
    Affiliations
    Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA
    Search for articles by this author
  • Guisheng Song
    Correspondence
    Corresponding authors. Address: Division of Gastroenterology, Hepatology and Nutrition, University of Minnesota, 516 Delaware Street SE, Minneapolis, MN 55455, USA (G. Song), or Key Laboratory of Chinese Medicine Resource and Compound Prescription, Ministry of Education, Hubei University of Chinese Medicine, 1 West Huangjiahu Road, Wuhan, Hubei Province 430065, China (G. Zheng).
    Affiliations
    Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA

    Key Laboratory of Chinese Medicine Resource and Compound Prescription, Ministry of Education, Hubei University of Chinese Medicine, Wuhan, Hubei Province 430065, China

    Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455, USA
    Search for articles by this author
  • Author Footnotes
    † Tianpeng Zhang and Junjie Hu contributed equally to this work.
Published:September 12, 2018DOI:https://doi.org/10.1016/j.jhep.2018.08.026

      Highlights

      • Hepatic expression of miR-378 is significantly upregulated in fatty livers of mice and patients with NASH.
      • miR-378 is a potent inhibitor of AMPK signaling.
      • miR-378 facilitates an inflammatory pathway of NFκB-TNFα by targeting Prkag2.
      • miR-378 robustly promotes hepatic inflammation and fibrosis in dietary obese mice.
      • TNFα signaling is required for miR-378 to induce NASH progression.

      Background & Aims

      The progression of hepatosteatosis to non-alcoholic steatohepatitis (NASH) is a critical step in the pathogenesis of hepatocellular cancer. However, the underlying mechanism(s) for this progression is essentially unknown. This study was designed to determine the role of miR-378 in regulating NASH progression.

      Methods

      We used immunohistochemistry, luciferase assays and immunoblotting to study the role of miR-378 in modulating an inflammatory pathway. Wild-type mice kept on a high-fat diet (HFD) were injected with miR-378 inhibitors or a mini-circle expression system containing miR-378, to study loss and gain-of functions of miR-378.

      Results

      MiR-378 expression is increased in livers of dietary obese mice and patients with NASH. Further studies revealed that miR-378 directly targeted Prkag2 that encodes AMP-activated protein kinase γ 2 (AMPKγ2). AMPK signaling negatively regulates the NF-κB-TNFα inflammatory axis by increasing deacetylase activity of sirtuin 1. By targeting Prkag2, miR-378 reduced sirtuin 1 activity and facilitated an inflammatory pathway involving NF-κB-TNFα. In contrast, miR-378 knockdown induced expression of Prkag2, increased sirtuin 1 activity and blocked the NF-κB-TNFα axis. Additionally, knockdown of increased Prkag2 offset the inhibitory effects of miR-378 inhibitor on the NF-κB-TNFα axis, suggesting that AMPK signaling mediates the role of miR-378 in facilitating this inflammatory pathway. Liver-specific expression of miR-378 triggered the development of NASH and fibrosis by activating TNFα signaling. Ablation of TNFα in miR-378-treated mice impaired the ability of miR-378 to facilitate hepatic inflammation and fibrosis, suggesting that TNFα signaling is required for miR-378 to promote NASH.

      Conclusion

      MiR-378 plays a key role in the development of hepatic inflammation and fibrosis by positively regulating the NF-κB-TNFα axis. MiR-378 is a potential therapeutic target for the treatment of NASH.

      Lay summary

      The recent epidemic of obesity has been associated with a sharp rise in the incidence of non-alcoholic fatty liver disease (NAFLD). However, the underlying mechanism(s) remains poorly described and effective therapeutic approaches against NAFLD are lacking. The results establish that microRNA-378 facilitates the development of hepatic inflammation and fibrosis and suggests the therapeutic potential of microRNA-378 inhibitor for the treatment of NAFLD.

      Graphical abstract

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Hepatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

      Author names in bold designate shared co-first authorship

        • Le M.H.
        • Devaki P.
        • Ha N.B.
        • Jun D.W.
        • Te H.S.
        • Cheung R.C.
        • et al.
        Prevalence of non-alcoholic fatty liver disease and risk factors for advanced fibrosis and mortality in the United States.
        PLoS ONE. 2017; 12e0173499
        • Younossi Z.
        • Anstee Q.M.
        • Marietti M.
        • Hardy T.
        • Henry L.
        • Eslam M.
        • et al.
        Global burden of NAFLD and NASH: trends, predictions, risk factors and prevention.
        Nat Rev Gastroenterol Hepatol. 2018; 15: 11
        • Issa D.
        • Alkhouri N.
        Nonalcoholic fatty liver disease and hepatocellular carcinoma: new insights on presentation and natural history.
        Hepatobiliary Surg Nutr. 2017; 6: 401
        • Foog D.H.-S.
        • Kwok D.
        • Yu B.C.-Y.
        • Wong V.W.-S.
        Managing HCC in NAFLD. Curr Hepatol.
        Rep. 2017; 16: 374-381
        • Marrero J.A.
        • Fontana R.J.
        • Su G.L.
        • Conjeevaram H.S.
        • Emick D.M.
        • Lok A.S.
        NAFLD may be a common underlying liver disease in patients with hepatocellular carcinoma in the United States.
        Hepatology. 2002; 36: 1349-1354
        • Ali M.A.
        • Lacin S.
        • Abdel-Wahab R.
        • Uemura M.
        • Hassan M.
        • Rashid A.
        • et al.
        Nonalcoholic steatohepatitis-related hepatocellular carcinoma: is there a role for the androgen receptor pathway?.
        Onco Targets Ther. 2017; 10: 1403
        • Cholankeril G.
        • Patel R.
        • Khurana S.
        • Satapathy S.K.
        Hepatocellular carcinoma in non-alcoholic steatohepatitis: current knowledge and implications for management.
        World J Hepatol. 2017; 9: 533
        • Wree A.
        • Broderick L.
        • Canbay A.
        • Hoffman H.M.
        • Feldstein A.E.
        From NAFLD to NASH to cirrhosis—new insights into disease mechanisms.
        Nat Rev Gastroenterol Hepatol. 2013; 10: 627
        • Tiniakos D.G.
        • Vos M.B.
        • Brunt E.M.
        Nonalcoholic fatty liver disease: pathology and pathogenesis.
        Annu Rev Pathol Mech. 2010; 5: 145-171
        • Marra F.
        • Gastaldelli A.
        • Baroni G.S.
        • Tell G.
        • Tiribelli C.
        Molecular basis and mechanisms of progression of non-alcoholic steatohepatitis.
        Trends Mol Med. 2008; 14: 72-81
        • Ratziu V.
        • Harrison S.A.
        • Francque S.
        • Bedossa P.
        • Lehert P.
        • Serfaty L.
        • et al.
        Elafibranor, an agonist of the peroxisome proliferator-activated receptor α and δ, induces resolution of nonalcoholic steatohepatitis without fibrosis worsening.
        Gastroenterology. 2016; 150: 1147-1159
        • Souza-Mello V.
        Peroxisome proliferator-activated receptors as targets to treat non-alcoholic fatty liver disease.
        World J Hepatol. 2015; 7: 1012
        • Rolo A.P.
        • Teodoro J.S.
        • Palmeira C.M.
        Role of oxidative stress in the pathogenesis of nonalcoholic steatohepatitis.
        Free Radic Biol Med. 2012; 52: 59-69
        • Bartel D.
        MicroRNAs: genomics, biogenesis, mechanism, and function.
        Cell. 2004; 116: 281-297
        • Cheung O.
        • Puri P.
        • Eicken C.
        • Contos M.J.
        • Mirshahi F.
        • Maher J.W.
        • et al.
        Nonalcoholic steatohepatitis is associated with altered hepatic microRNA expression.
        Hepatology. 2008; 48: 1810-1820
        • Becker P.
        • Niesler B.
        • Tschopp O.
        • Berr F.
        • Canbay A.
        • Dandekar T.
        • et al.
        MicroRNAs as mediators in the pathogenesis of non-alcoholic fatty liver disease and steatohepatitis.
        Zeitschrift für Gastroenterologie. 2014; 52: 1-27
        • Li M.
        • Tang Y.
        • Wu L.
        • Mo F.
        • Wang X.
        • Li H.
        • et al.
        The hepatocyte-specific HNF4α/miR-122 pathway contributes to iron overload–mediated hepatic inflammation.
        Blood. 2017; 130: 1041-1051
        • Csak T.
        • Bala S.
        • Lippai D.
        • Kodys K.
        • Catalano D.
        • Iracheta-Vellve A.
        • et al.
        MicroRNA-155 deficiency attenuates liver steatosis and fibrosis without reducing inflammation in a mouse model of steatohepatitis.
        PLoS ONE. 2015; 10e0129251
        • Takahashi Y.
        • Soejima Y.
        • Fukusato T.
        Animal models of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis.
        World J Gastro. 2012; 18: 2300
        • Santhekadur P.K.
        • Kumar D.P.
        • Sanyal A.J.
        Preclinical models of nonalcoholic fatty liver disease.
        J Hepatol. 2018; 68: 230-237
        • Zhang T.
        • Zhao X.
        • Steer C.J.
        • Yan G.
        • Song G.
        A negative feedback loop between microRNA-378 and Nrf1 promotes the development of hepatosteatosis in mice treated with a high fat diet.
        Metabolism. 2018; 85: 183-191
        • Carrer M.
        • Liu N.
        • Grueter C.E.
        • Williams A.H.
        • Frisard M.I.
        • Hulver M.W.
        • et al.
        Control of mitochondrial metabolism and systemic energy homeostasis by microRNAs 378 and 378*.
        PNAS. 2012; 109: 15330-15335
        • Jeon T.I.
        • Park J.W.
        • Ahn J.
        • Jung C.H.
        • Ha T.Y.
        Fisetin protects against hepatosteatosis in mice by inhibiting miR-378.
        Mol Nutr Food Res. 2013; 57: 1931-1937
        • Liu W.
        • Cao H.
        • Ye C.
        • Chang C.
        • Lu M.
        • Jing Y.
        • et al.
        Hepatic miR-378 targets p110α and controls glucose and lipid homeostasis by modulating hepatic insulin signalling.
        Nat Commun. 2014; 5: 5684
        • Jayandharan G.R.
        • Zhong L.
        • Sack B.K.
        • Rivers A.E.
        • Li M.
        • Li B.
        • et al.
        Optimized Adeno-Associated Virus (AAV)-protein phosphatase-5 helper viruses for efficient liver transduction by single-stranded AAV vectors: therapeutic expression of factor IX at reduced vector doses.
        Hum Gene Ther. 2010; 21: 271-283
        • Wang B.
        • Majumder S.
        • Nuovo G.
        • Kutay H.
        • Volinia S.
        • Patel T.
        • et al.
        Role of microRNA-155 at early stages of hepatocarcinogenesis induced by choline-deficient and amino acid-defined diet in C57BL/6 mice.
        Hepatology. 2009; 50: 1152-1161
        • Gomez-Lechon M.J.
        • Donato M.T.
        • Martínez-Romero A.
        • Jiménez N.
        • Castell J.V.
        • O’Connor J.-E.
        A human hepatocellular in vitro model to investigate steatosis.
        Chemico-Bio Inter. 2007; 165: 106-116
        • Lewis B.P.
        • Burge C.B.
        • Bartel D.P.
        Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets.
        Cell. 2005; 120: 15-20
        • Gao G.
        • Fernandez C.S.
        • Stapleton D.
        • Auster A.S.
        • Widmer J.
        • Dyck J.R.
        • et al.
        Non-catalytic-and-subunit isoforms of the 5-AMP-activated protein kinase.
        J Biol Chem. 1996; 271: 8675-8681
        • Salminen A.
        • Hyttinen J.M.
        • Kaarniranta K.
        AMP-activated protein kinase inhibits NF-κB signaling and inflammation: impact on healthspan and lifespan.
        Int J Mol Med. 2011; 89: 667-676
        • Cantó C.
        • Gerhart-Hines Z.
        • Feige J.N.
        • Lagouge M.
        • Noriega L.
        • Milne J.C.
        • et al.
        AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity.
        Nature. 2009; 458: 1056-1060
        • Yeung F.
        • Hoberg J.E.
        • Ramsey C.S.
        • Keller M.D.
        • Jones D.R.
        • Frye R.A.
        • et al.
        Modulation of NF-κB-dependent transcription and cell survival by the SIRT1 deacetylase.
        EMBO J. 2004; 23: 2369-2380
        • Mihaylova M.M.
        • Shaw R.J.
        The AMPK signalling pathway coordinates cell growth, autophagy and metabolism.
        Nat Cell Biol. 2011; 13: 1016-1023
        • Mayrhofer P.
        • Schleef M.
        • Jechlinger W.
        Use of minicircle plasmids for gene therapy.
        Cancer Gene Ther: Springer. 2009; : 87-104
        • Dara L.
        The receptor interacting protein kinases in the liver.
        Semin Liver Dis. 2018; 2018: 073-086
        • Kamimura D.
        • Ishihara K.
        • Hirano T.
        IL-6 signal transduction and its physiological roles: the signal orchestration model.
        Rev Physiol Biochem Pharmacol: Springer. 2003; : 1-38
        • Luckett-Chastain L.
        • Gallucci R.
        Interleukin (IL)-6 modulates transforming growth factor-β expression in skin and dermal fibroblasts from IL-6-deficient mice.
        Br J Dermatol. 2009; 161: 237-248
        • Wang X.
        • Zheng Z.
        • Caviglia J.M.
        • Corey K.E.
        • Herfel T.M.
        • Cai B.
        • et al.
        Hepatocyte TAZ/WWTR1 promotes inflammation and fibrosis in nonalcoholic steatohepatitis.
        Cell Metab. 2016; 24: 848-862
        • Xu Z.
        • Chen L.
        • Leung L.
        • Yen T.B.
        • Lee C.
        • Chan J.Y.
        Liver-specific inactivation of the Nrf1 gene in adult mouse leads to nonalcoholic steatohepatitis and hepatic neoplasia.
        PNAS. 2005; 102: 4120-4125
        • Sahai A.
        • Malladi P.
        • Melin-Aldana H.
        • Green R.M.
        • Whitington P.F.
        Upregulation of osteopontin expression is involved in the development of nonalcoholic steatohepatitis in a dietary murine model.
        Am J Physiol Gastrointest Liver Physiol. 2004; 287: G264-G273
        • Matsumoto M.
        • Hada N.
        • Sakamaki Y.
        • Uno A.
        • Shiga T.
        • Tanaka C.
        • et al.
        An improved mouse model that rapidly develops fibrosis in non-alcoholic steatohepatitis.
        Int J Clin Exp Pathol. 2013; 94: 93-103
        • Smith B.K.
        • Marcinko K.
        • Desjardins E.M.
        • Lally J.S.
        • Ford R.J.
        • Steinberg G.R.
        Treatment of nonalcoholic fatty liver disease: role of AMPK.
        Am J Physiol Endocrinol Metab. 2016; 311: E730-E740