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RIP1 kinase activity promotes steatohepatitis through mediating cell death and inflammation in macrophages

Cell Death & Differentiation volume 28, pages 1418–1433 (2021)Cite this article

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Abstract

Hepatocyte cell death and liver inflammation have been well recognized as central characteristics of nonalcoholic steatohepatitis (NASH), however, the underlying molecular basis remains elusive. The kinase receptor-interacting protein 1 (RIP1) is a multitasking molecule with distinct functions in regulating apoptosis, necroptosis, and inflammation. Dissecting the role of RIP1 distinct functions in different pathophysiology has absorbed huge research enthusiasm. Wild-type and RIP1 kinase-dead (Rip1K45A/K45A) mice were fed with high-fat diet (HFD) to investigate the role of RIP1 kinase activity in the pathogenesis of NASH. Rip1K45A/K45A mice exhibited significantly alleviated NASH phenotype of hepatic steatosis, liver damage, fibrosis as well as reduced hepatic cell death and inflammation compared to WT mice. Our results also indicated that both in vivo lipotoxicity and in vitro saturated fatty acids (palmitic acid) treatment were able to induce the kinase activation of RIP1 in liver macrophages. RIP1 kinase was required for mediating inflammasome activation, apoptotic and necrotic cell death induced by palmitic acid in both bone marrow-derived macrophage and mouse primary Kupffer cells. Results from chimeric mice established through lethal irradiation and bone marrow transplantation further confirmed that the RIP1 kinase in hematopoietic-derived macrophages contributed mostly to the disease progression in NASH. Consistent with murine models, we also found that RIP1 kinase was markedly activated in human NASH, and the kinase activation mainly occurred in liver macrophages as indicated by immunofluorescence double staining. In summary, our study indicated that RIP1 kinase was phosphorylated and activated mainly in liver macrophages in both experimental and clinical NASH. We provided direct genetic evidence that the kinase activity of RIP1 especially in hematopoietic-derived macrophages contributes to the pathogenesis of NASH, through mediating inflammasome activation and cell death induction. Macrophage RIP1 kinase represents a specific and potential therapeutic target for NASH.

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Fig. 1: RIP1 kinase prompted the liver injury, steatosis, and fibrosis in HFD-induced steatohepatitis.
Fig. 2: RIP kinase activity contributed to the hepatic cell death, inflammatory cell infiltration and liver inflammation in HFD-induced steatohepatitis.
Fig. 3: RIP1 kinase activity in hematopoietic-derived macrophages was crucial for the disease progression of steatohepatitis.
Fig. 4: RIP1 kinase activity in hematopoietic-derived macrophages contributed to the liver inflammation and hepatic cell death induction in steatohepatitis.
Fig. 5: RIP1 kinase mediated saturated fatty acid PA-induced cell death and inflammasome activation in both bone marrow-derived macrophages and primary Kupffer cells.
Fig. 6: RIP1 kinase was significantly activated in hepatic macrophages in human NASH.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China under Grant 31970897 and 21677076, Outstanding Youth Foundation of Jiangsu Province (BK20190069), the Fundamental Research Funds for the Central Universities No. 30919011102, Qing Lan Project of Jiangsu Province.

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Affiliations

  1. Center for Molecular Metabolism, Nanjing University of Science & Technology, 200 Xiaolingwei Street, 210094, Nanjing, China

    Liang Tao, Yuguo Yi, Jiapeng Jie, Weigao Zhang, Qian Xu, Yang Li, Zhao Ding, Junsong Wang, Jianfa Zhang & Dan Weng

  2. Department of Laboratory Medicine, Nanjing Drum Tower Hospital, Nanjing University Medical School, Nanjing, 210008, Jiangsu, China

    Yuxin Chen

  3. CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 100864, Shanghai, China

    Haibing Zhang & Qiurong Ding

  4. Program in Innate Immunity, Department of Medicine, Division of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, MA, 01605, USA

    Pontus Orning & Egil Lien

  5. Centre of Molecular Inflammation Research, Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, 7491, Trondheim, Norway

    Pontus Orning & Egil Lien

  6. Department of Infectious diseases, Nanjing Drum Tower Hospital, Nanjing University Medical School, Nanjing, 210008, Jiangsu, China

    Chao Wu

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Tao, L., Yi, Y., Chen, Y. et al. RIP1 kinase activity promotes steatohepatitis through mediating cell death and inflammation in macrophages. Cell Death Differ 28, 1418–1433 (2021). https://doi.org/10.1038/s41418-020-00668-w

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