Advertisement

IL-6/STAT3 axis dictates the PNPLA3-mediated susceptibility to non-alcoholic fatty liver disease

  • Author Footnotes
    † These authors contributed equally.
    Jiwoon Park
    Footnotes
    † These authors contributed equally.
    Affiliations
    Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY, USA

    Department of Physiology, Biophysics and Systems Biology, Weill Cornell Medicine, New York, NY, USA

    The HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medicine, New York, NY, USA
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally.
    Yuanyuan Zhao
    Footnotes
    † These authors contributed equally.
    Affiliations
    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally.
    Fan Zhang
    Footnotes
    † These authors contributed equally.
    Affiliations
    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally.
    Shaoyan Zhang
    Footnotes
    † These authors contributed equally.
    Affiliations
    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally.
    Andrew C. Kwong
    Footnotes
    † These authors contributed equally.
    Affiliations
    Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY, USA

    The Biological and Biomedical Sciences Program, Harvard Medical School, Boston, MA, USA
    Search for articles by this author
  • Yujie Zhang
    Affiliations
    Department of Biomedical Sciences, College of Medicine, Florida State University, Tallahassee, FL, USA
    Search for articles by this author
  • Hans-Heinrich Hoffmann
    Affiliations
    Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY, USA
    Search for articles by this author
  • Leila Bushweller
    Affiliations
    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Xin Wu
    Affiliations
    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Alison W. Ashbrook
    Affiliations
    Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY, USA
    Search for articles by this author
  • Branko Stefanovic
    Affiliations
    Department of Biomedical Sciences, College of Medicine, Florida State University, Tallahassee, FL, USA
    Search for articles by this author
  • Shuyang Chen
    Affiliations
    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Andrea D. Branch
    Affiliations
    Division of Liver Diseases, Icahn School of Medicine at Mount Sinai, New York, NY, USA
    Search for articles by this author
  • Christopher E. Mason
    Affiliations
    Department of Physiology, Biophysics and Systems Biology, Weill Cornell Medicine, New York, NY, USA

    The HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medicine, New York, NY, USA
    Search for articles by this author
  • Jae U. Jung
    Affiliations
    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Charles M. Rice
    Correspondence
    Corresponding authors. Addresses: 1230 York Ave, New York, NY 10065, USA. Tel.: +1-212-327-7046; Fax: +1-212-327-7048
    Affiliations
    Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY, USA
    Search for articles by this author
  • Xianfang Wu
    Correspondence
    9500 Euclid Avenue/NE20, Cleveland, OH 44195, USA. Tel.: +1-216-445-1458; Fax: +1-216-444-0512.
    Affiliations
    Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY, USA

    Infection Biology Program and Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally.
Published:August 29, 2022DOI:https://doi.org/10.1016/j.jhep.2022.08.022

      Highlights

      • A hPSC-derived multicellular liver culture mimicking liver composition was developed.
      • Multicellular liver culture recapitulates many key features of NAFLD development.
      • Multicellular liver cultures harbouring PNPLA3I148M enhance susceptibility to NAFLD.
      • Elevating IL-6/STAT3 signalling promotes PNPLA3I148M-induced NAFLD progression.
      • Blocking trans-signalling by sgp130Fc protects against PNPLA3I148M-induced NAFLD progression.

      Background & Aims

      A number of genetic polymorphisms have been associated with susceptibility to or protection against non-alcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain unknown. Here, we focused on the rs738409 C>G single nucleotide polymorphism (SNP), which produces the I148M variant of patatin-like phospholipase domain-containing protein 3 (PNPLA3) and is strongly associated with NAFLD.

      Methods

      To enable mechanistic dissection, we developed a human pluripotent stem cell (hPSC)-derived multicellular liver culture by incorporating hPSC-derived hepatocytes, hepatic stellate cells, and macrophages. We first applied this liver culture to model NAFLD by utilising a lipotoxic milieu reflecting the circulating levels of disease risk factors in affected individuals. We then created an isogenic pair of liver cultures differing only at rs738049 and compared NAFLD phenotype development.

      Results

      Our hPSC-derived liver culture recapitulated many key characteristics of NAFLD development and progression including lipid accumulation and oxidative stress, inflammatory response, and stellate cell activation. Under the lipotoxic conditions, the I148M variant caused the enhanced development of NAFLD phenotypes. These differences were associated with elevated IL-6/signal transducer and activator of transcription 3 (STAT3) activity in liver cultures, consistent with transcriptomic data of liver biopsies from individuals carrying the rs738409 SNP. Dampening IL-6/STAT3 activity alleviated the I148M-mediated susceptibility to NAFLD, whereas boosting it in wild-type liver cultures enhanced NAFLD development. Finally, we attributed this elevated IL-6/STAT3 activity in liver cultures carrying the rs738409 SNP to increased NF-κB activity.

      Conclusions

      Our study thus reveals a potential causal link between elevated IL-6/STAT3 activity and 148M-mediated susceptibility to NAFLD.

      Impact and implications

      An increasing number of genetic variants manifest in non-alcoholic fatty liver disease (NAFLD) development and progression; however, the underlying mechanisms remain elusive. To study these variants in human-relevant systems, we developed an induced pluripotent stem cell-derived multicellular liver culture and focused on a common genetic variant (i.e. rs738409 in PNPLA3). Our findings not only provide mechanistic insight, but also a potential therapeutic strategy for NAFLD driven by this genetic variant in PNPLA3. Our liver culture is therefore a useful platform for exploring genetic variants in NAFLD development.

      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

        • Loomba R.
        • Friedman S.L.
        • Shulman G.I.
        Mechanisms and disease consequences of nonalcoholic fatty liver disease.
        Cell. 2021; 184: 2537-2564
        • Trépo E.
        • Valenti L.
        Update on NAFLD genetics: from new variants to the clinic.
        J Hepatol. 2020; 72: 1196-1209
        • Friedman S.L.
        • Neuschwander-Tetri B.A.
        • Rinella M.
        • Sanyal A.J.
        Mechanisms of NAFLD development and therapeutic strategies.
        Nat Med. 2018; 24: 908-922
        • Romeo S.
        • Kozlitina J.
        • Xing C.
        • Pertsemlidis A.
        • Cox D.
        • Pennacchio L.A.
        • et al.
        Genetic variation in PNPLA3 confers susceptibility to nonalcoholic fatty liver disease.
        Nat Genet. 2008; 40: 1461-1465
        • Abul-Husn N.S.
        • Cheng X.
        • Li A.H.
        • Xin Y.
        • Schurmann C.
        • Stevis P.
        • et al.
        A protein-truncating HSD17B13 variant and protection from chronic liver disease.
        N Engl J Med. 2018; 378: 1096-1106
        • Chitturi S.
        • Abeygunasekera S.
        • Farrell G.C.
        • Holmes-Walker J.
        • Hui J.M.
        • Fung C.
        • et al.
        NASH and insulin resistance: insulin hypersecretion and specific association with the insulin resistance syndrome.
        Hepatology. 2002; 35: 373-379
        • Tavares De Almeida I.
        • Cortez-Pinto H.
        • Fidalgo G.
        • Rodrigues D.
        • Camilo M.E.
        Plasma total and free fatty acids composition in human non-alcoholic steatohepatitis.
        Clin Nutr. 2002; 21: 219-223
        • Sookoian S.
        • Pirola C.J.
        Meta-analysis of the influence of I148M variant of patatin-like phospholipase domain containing 3 gene (PNPLA3) on the susceptibility and histological severity of nonalcoholic fatty liver disease.
        Hepatology. 2011; 53: 1883-1894
        • Govaere O.
        • Cockell S.
        • Tiniakos D.
        • Queen R.
        • Younes R.
        • Vacca M.
        • et al.
        Transcriptomic profiling across the nonalcoholic fatty liver disease spectrum reveals gene signatures for steatohepatitis and fibrosis.
        Sci Transl Med. 2020; 12eaba4448
        • Tilson S.G.
        • Morell C.M.
        • Lenaerts A.-S.
        • Park S.B.
        • Hu Z.
        • Jenkins B.
        • et al.
        Modeling PNPLA3-associated NAFLD using human-induced pluripotent stem cells.
        Hepatology. 2021; 74: 2998-3017
        • Arab J.P.
        • Arrese M.
        • Trauner M.
        Recent insights into the pathogenesis of nonalcoholic fatty liver disease.
        Annu Rev Pathol. 2018; 13: 321-350
        • Coll M.
        • Perea L.
        • Boon R.
        • Leite S.B.
        • Vallverdú J.
        • Mannaerts I.
        • et al.
        Generation of hepatic stellate cells from human pluripotent stem cells enables in vitro modeling of liver fibrosis.
        Cell Stem Cell. 2018; 23: 101-113.e107
        • El Taghdouini A.
        • Najimi M.
        • Sancho-Bru P.
        • Sokal E.
        • van Grunsven L.A.
        In vitro reversion of activated primary human hepatic stellate cells.
        Fibrogenesis Tissue Repair. 2015; 8: 14
        • Si-Tayeb K.
        • Lemaigre F.P.
        • Duncan S.A.
        Organogenesis and development of the liver.
        Dev Cel. 2010; 18: 175-189
        • Bilzer M.
        • Roggel F.
        • Gerbes A.L.
        Role of Kupffer cells in host defense and liver disease.
        Liver Int. 2006; 26: 1175-1186
        • Sharifnia T.
        • Antoun J.
        • Verriere T.G.C.
        • Suarez G.
        • Wattacheril J.
        • Wilson K.T.
        • et al.
        Hepatic TLR4 signaling in obese NAFLD.
        Am J Physiol Gastrointest Liver Physiol. 2015; 309: G270-G278
        • Lin H.V.
        • Accili D.
        Hormonal regulation of hepatic glucose production in health and disease.
        Cell Metabol. 2011; 14: 9-19
        • Feaver R.E.
        • Cole B.K.
        • Lawson M.J.
        • Hoang S.A.
        • Marukian S.
        • Blackman B.R.
        • et al.
        Development of an in vitro human liver system for interrogating nonalcoholic steatohepatitis.
        JCI Insight. 2016; 1e90954
        • Johansen J.S.
        • Christoffersen P.
        • Møller S.
        • Price P.A.
        • Henriksen J.H.
        • Garbarsch C.
        • et al.
        Serum YKL-40 is increased in patients with hepatic fibrosis.
        J.Hepatol. 2000; 32: 911-920
        • Tanaka T.
        • Narazaki M.
        • Kishimoto T.
        IL-6 in inflammation, immunity, and disease.
        Cold Spring Harb Perspect Biol. 2014; 6: a016295
        • Wieckowska A.
        • Papouchado B.G.
        • Li Z.
        • Lopez R.
        • Zein N.N.
        • Feldstein A.E.
        Increased hepatic and circulating interleukin-6 levels in human nonalcoholic steatohepatitis.
        Am J Gastroenterol. 2008; 103: 1372-1379
        • Pirazzi C.
        • Valenti L.
        • Motta B.M.
        • Pingitore P.
        • Hedfalk K.
        • Mancina R.M.
        • et al.
        PNPLA3 has retinyl-palmitate lipase activity in human hepatic stellate cells.
        Hum Mol Genet. 2014; 23: 4077-4085
        • BasuRay S.
        • Smagris E.
        • Cohen J.C.
        • Hobbs H.H.
        The PNPLA3 variant associated with fatty liver disease (I148M) accumulates on lipid droplets by evading ubiquitylation.
        Hepatology. 2017; 66: 1111-1124
        • Suppli M.P.
        • Rigbolt K.T.G.
        • Veidal S.S.
        • Heebøll S.
        • Eriksen P.L.
        • Demant M.
        • et al.
        Hepatic transcriptome signatures in patients with varying degrees of nonalcoholic fatty liver disease compared with healthy normal-weight individuals.
        Am J Physiol Gastrointest Liver Physiol. 2019; 316: G462-G472
        • Luukkonen P.K.
        • Nick A.
        • Hölttä-Vuori M.
        • Thiele C.
        • Isokuortti E.
        • Lallukka-Brück S.
        • et al.
        Human PNPLA3-I148M variant increases hepatic retention of polyunsaturated fatty acids.
        JCI Insight. 2019; 4e127902
        • Giraldez M.D.
        • Carneros D.
        • Garbers C.
        • Rose-John S.
        • Bustos M.
        New insights into IL-6 family cytokines in metabolism, hepatology and gastroenterology.
        Nat Rev Gastroenterol Hepatol. 2021; 18: 787-803
        • Jones S.A.
        • Jenkins B.J.
        Recent insights into targeting the IL-6 cytokine family in inflammatory diseases and cancer.
        Nat Rev Immunol. 2018; 18: 773-789
        • Garbers C.
        • Heink S.
        • Korn T.
        • Rose-John S.
        Interleukin-6: designing specific therapeutics for a complex cytokine.
        Nat Rev Drug Discov. 2018; 17: 395-412
        • Collin de l’Hortet A.
        • Takeishi K.
        • Guzman-Lepe J.
        • Morita K.
        • Achreja A.
        • Popovic B.
        • et al.
        Generation of human fatty livers using custom-engineered induced pluripotent stem cells with modifiable SIRT1 metabolism.
        Cell Metabol. 2019; 30: 385-401.e389
        • Matthews V.B.
        • Allen T.L.
        • Risis S.
        • Chan M.H.S.
        • Henstridge D.C.
        • Watson N.
        • et al.
        Interleukin-6-deficient mice develop hepatic inflammation and systemic insulin resistance.
        Diabetologia. 2010; 53: 2431-2441
        • Schreiber S.
        • Aden K.
        • Bernardes J.P.
        • Conrad C.
        • Tran F.
        • Höper H.
        • et al.
        Therapeutic interleukin-6 trans-signaling inhibition by olamkicept (sgp130Fc) in patients with active inflammatory bowel disease.
        Gastroenterology. 2021; 160: 2354-2366 e2311
        • Bergmann J.
        • Müller M.
        • Baumann N.
        • Reichert M.
        • Heneweer C.
        • Bolik J.
        • et al.
        IL-6 trans-signaling is essential for the development of hepatocellular carcinoma in mice.
        Hepatology. 2017; 65: 89-103
        • Bekkering S.
        • Quintin J.
        • Joosten L.A.
        • van der Meer J.W.
        • Netea M.G.
        • Riksen N.P.
        Oxidized low-density lipoprotein induces long-term proinflammatory cytokine production and foam cell formation via epigenetic reprogramming of monocytes.
        Arterioscler Thromb Vasc Biol. 2014; 34: 1731-1738
        • Colak Y.
        • Senates E.
        • Ozturk O.
        • Doganay H.L.
        • Coskunpinar E.
        • Oltulu Y.M.
        • et al.
        Association of serum lipoprotein-associated phospholipase A2 level with nonalcoholic fatty liver disease.
        Metab Syndr Relat Disord. 2012; 10: 103-109
        • Triggiani M.
        • Granata F.
        • Oriente A.
        • Gentile M.
        • Petraroli A.
        • Balestrieri B.
        • et al.
        Secretory phospholipases A2 induce cytokine release from blood and synovial fluid monocytes.
        Eur J Immunol. 2002; 32: 67-76