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Glycogen synthase kinase 3β promotes liver innate immune activation by restraining AMP-activated protein kinase activation

  • Author Footnotes
    † These authors contributed equally to the work.
    Haoming Zhou
    Footnotes
    † These authors contributed equally to the work.
    Affiliations
    Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at the University of California, Los Angeles, CA, USA

    Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
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  • Author Footnotes
    † These authors contributed equally to the work.
    Han Wang
    Footnotes
    † These authors contributed equally to the work.
    Affiliations
    Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at the University of California, Los Angeles, CA, USA

    Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
    Search for articles by this author
  • Ming Ni
    Affiliations
    Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at the University of California, Los Angeles, CA, USA

    Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
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  • Shi Yue
    Affiliations
    Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
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  • Yongxiang Xia
    Affiliations
    Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
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  • Ronald W. Busuttil
    Affiliations
    Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at the University of California, Los Angeles, CA, USA
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  • Jerzy W. Kupiec-Weglinski
    Affiliations
    Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at the University of California, Los Angeles, CA, USA
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  • Ling Lu
    Affiliations
    Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
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  • Xuehao Wang
    Correspondence
    Corresponding authors. Addresses: Department of Liver Surgery, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing 210029, China. Tel.: +86 25 68136053; fax: +86 25 84630769 (X. Wang). Dumont-UCLA Transplant Center 77-120 CHS, 10833 Le Conte Ave, Los Angeles, CA 90095, USA. Tel.: +1 310 8259426; fax: +1 310 2672367 (Y. Zhai).
    Affiliations
    Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu Province, China
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  • Yuan Zhai
    Correspondence
    Corresponding authors. Addresses: Department of Liver Surgery, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing 210029, China. Tel.: +86 25 68136053; fax: +86 25 84630769 (X. Wang). Dumont-UCLA Transplant Center 77-120 CHS, 10833 Le Conte Ave, Los Angeles, CA 90095, USA. Tel.: +1 310 8259426; fax: +1 310 2672367 (Y. Zhai).
    Affiliations
    Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at the University of California, Los Angeles, CA, USA
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally to the work.
Published:February 13, 2018DOI:https://doi.org/10.1016/j.jhep.2018.01.036

      Highlights

      • Myeloid Gsk3β promotes liver pro-inflammatory immune responses to ischemia reperfusion.
      • Active Gsk3β inhibits AMPK activation and SHP induction in activated macrophages.
      • Gsk3β regulates AMPK/SHP independent of its S9 phosphorylation status.

      Background & Aims

      Glycogen synthase kinase 3β (Gsk3β [Gsk3b]) is a ubiquitously expressed kinase with distinctive functions in different types of cells. Although its roles in regulating innate immune activation and ischaemia and reperfusion injuries (IRIs) have been well documented, the underlying mechanisms remain ambiguous, in part because of the lack of cell-specific tools in vivo.

      Methods

      We created a myeloid-specific Gsk3b knockout (KO) strain to study the function of Gsk3β in macrophages in a murine liver partial warm ischaemia model.

      Results

      Compared with controls, myeloid Gsk3b KO mice were protected from IRI, with diminished proinflammatory but enhanced anti-inflammatory immune responses in livers. In bone marrow-derived macrophages, Gsk3β deficiency resulted in an early reduction of Tnf gene transcription but sustained increase of Il10 gene transcription on Toll-like receptor 4 stimulation in vitro. These effects were associated with enhanced AMP-activated protein kinase (AMPK) activation, which led to an accelerated and higher level of induction of the novel innate immune negative regulator small heterodimer partner (SHP [Nr0b2]). The regulatory function of Gsk3β on AMPK activation and SHP induction was confirmed in wild-type bone marrow-derived macrophages with a Gsk3 inhibitor. Furthermore, we found that this immune regulatory mechanism was independent of Gsk3β Ser9 phosphorylation and the phosphoinositide 3-kinase–Akt signalling pathway. In vivo, myeloid Gsk3β deficiency facilitated SHP upregulation by ischaemia–reperfusion in liver macrophages. Treatment of Gsk3b KO mice with either AMPK inhibitor or SHP small interfering RNA before the onset of liver ischaemia restored liver proinflammatory immune activation and IRI in these otherwise protected hosts. Additionally, pharmacological activation of AMPK protected wild-type mice from liver IRI, with reduced proinflammatory immune activation. Inhibition of the AMPK–SHP pathway by liver ischaemia was demonstrated in tumour resection patients.

      Conclusions

      Gsk3β promotes innate proinflammatory immune activation by restraining AMPK activation.

      Lay summary

      Glycogen synthase kinase 3β promotes macrophage inflammatory activation by inhibiting the immune regulatory signalling of AMP-activated protein kinase and the induction of small heterodimer partner. Therefore, therapeutic targeting of glycogen synthase kinase 3β enhances innate immune regulation and protects liver from ischaemia and reperfusion injury.

      Graphical abstract

      Keywords

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