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Gasdermin D plays a key role as a pyroptosis executor of non-alcoholic steatohepatitis in humans and mice

  • Author Footnotes
    † These authors contributed equally to this work.
    Bing Xu
    Footnotes
    † These authors contributed equally to this work.
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Author Footnotes
    † These authors contributed equally to this work.
    Mingzuo Jiang
    Footnotes
    † These authors contributed equally to this work.
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Author Footnotes
    † These authors contributed equally to this work.
    Yi Chu
    Footnotes
    † These authors contributed equally to this work.
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Author Footnotes
    † These authors contributed equally to this work.
    Weijie Wang
    Footnotes
    † These authors contributed equally to this work.
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Di Chen
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Xiaowei Li
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Zhao Zhang
    Affiliations
    Lintong Aerial Medical Evaluation and Training Center of Air Force, PLA, Xi’an, Shaanxi, MI 710032, China
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  • Di Zhang
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Daiming Fan
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Yongzhan Nie
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Feng Shao
    Affiliations
    National Institute of Biological Sciences, Number 7 Science Park Road, Zhongguancun Life Science Park, Beijing, MI 102206, China
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  • Kaichun Wu
    Correspondence
    Corresponding authors. Address: State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Number 169 Changle West Road, Xi’an, Shaanxi Province 710032, China.
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
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  • Jie Liang
    Correspondence
    Corresponding authors. Address: State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Number 169 Changle West Road, Xi’an, Shaanxi Province 710032, China.
    Affiliations
    State Key Laboratory of Cancer Biology, National Clinical Research Center for Digestive Diseases and Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, Shaanxi, MI 710032, China
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally to this work.
Published:December 19, 2017DOI:https://doi.org/10.1016/j.jhep.2017.11.040

      Highlights

      • Hepatic N-terminal cleavage fragments of GSDMD (GSDMD-N) are associated with lobular inflammation and hepatic ballooning.
      • GSDMD-N is a potential biomarker for the diagnosis of non-alcoholic steatohepatitis.
      • GSDMD plays a key role in steatohepatitis by mediating macrophage infiltration, NF-ĸB activation and lipogenesis.

      Background & Aims

      Gasdermin D (GSDMD)-executed programmed necrosis is involved in inflammation and controls interleukin (IL)-1β release. However, the role of GSDMD in non-alcoholic steatohepatitis (NASH) remains unclear. We investigated the role of GSDMD in the pathogenesis of steatohepatitis.

      Methods

      Human liver tissues from patients with non-alcoholic fatty liver disease (NAFLD) and control individuals were obtained to evaluate GSDMD expression. Gsdmd knockout (Gsdmd−/−) mice, obese db/db mice and their wild-type (WT) littermates were fed with methionine-choline deficient (MCD) or control diet to induce steatohepatitis. The Gsdmd−/− and WT mice were also used in a high-fat diet (HFD)-induced NAFLD model. In addition, Alb-Cre mice were administered an adeno-associated virus (AAV) vector that expressed the gasdermin-N domain (AAV9-FLEX-GSDMD-N) and were fed with either MCD or control diet for 10 days.

      Results

      GSDMD and its pyroptosis-inducing fragment GSDMD-N were upregulated in liver tissues of human NAFLD/NASH. Importantly, hepatic GSDMD-N protein levels were significantly higher in human NASH and correlated with the NAFLD activity score and fibrosis. GSDMD-N remained a potential biomarker for the diagnosis of NASH. MCD-fed Gsdmd−/− mice exhibit decreased severity of steatosis and inflammation compared with WT littermates. GSDMD was associated with the secretion of pro-inflammatory cytokines (IL-1β, TNF-α, and MCP-1 [CCL2]) and persistent activation of the NF-ĸB signaling pathway. Gsdmd−/− mice showed lower steatosis, mainly because of reduced expression of the lipogenic gene Srebp1c (Srebf1) and upregulated expression of lipolytic genes, including Pparα, Aco [Klk15], Lcad [Acadl], Cyp4a10 and Cyp4a14. Alb-Cre mice administered with AAV9-FLEX-GSDMD-N showed significantly aggravated steatohepatitis when fed with MCD diet.

      Conclusion

      As an executor of pyroptosis, GSDMD plays a key role in the pathogenesis of steatohepatitis, by controlling cytokine secretion, NF-ĸB activation, and lipogenesis.

      Lay summary

      Non-alcoholic fatty liver disease has become one of the most feared chronic liver diseases, because it is the most rapidly growing indication for adult liver transplantation and a major cause of hepatocellular carcinoma. However, the mechanisms involved in the transformation of simple steatosis to steatohepatitis remain unclear. Herein, we show that gasdermin D driven pyroptosis is prominent in patients with non-alcoholic steatohepatitis (NASH), and gasdermin-N domain remains a potential biomarker for the diagnosis of NASH. Gasdermin D plays a key role in the pathogenesis of NASH by regulating lipogenesis, the inflammatory response, and the NF-ĸB signaling pathway, revealing potential treatment targets for NASH in humans.

      Graphical abstract

      Keywords

      Linked Article

      • Pyroptosis: An inflammatory link between NAFLD and NASH with potential therapeutic implications
        Journal of HepatologyVol. 68Issue 4
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          Non-alcoholic fatty liver disease (NAFLD) represents a major health problem worldwide because of its high and rising prevalence, its association with cardiovascular disease, and its link with an increased risk of developing cirrhosis and hepatocellular carcinoma (HCC). NAFLD is classified into different degrees, from simple steatosis (overall 20–30% prevalence), which is considered “benign”, to steatohepatitis (NASH: 2–5% prevalence) and fibrosis.1,2 The major risk factors of NAFLD include metabolic syndrome (i.e.
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