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MLKL-mediated endothelial necroptosis drives vascular damage and mortality in systemic inflammatory response syndrome

Cellular & Molecular Immunology volume 21, pages 1309–1321 (2024)Cite this article

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Abstract

The hypersecretion of cytokines triggers life-threatening systemic inflammatory response syndrome (SIRS), leading to multiple organ dysfunction syndrome (MODS) and mortality. Although both coagulopathy and necroptosis have been identified as important factors in the pathogenesis of SIRS, the specific cell types that undergo necroptosis and the interrelationships between coagulopathy and necroptosis remain unclear. In this study, we utilized visualization analysis via intravital microscopy to demonstrate that both anticoagulant heparin and nonanticoagulant heparin (NAH) pretreatment protect mice against TNF-α-induced mortality in SIRS. Moreover, the deletion of Mlkl or Ripk3 resulted in decreased coagulation and reduced mortality in TNF-α-induced SIRS. These findings suggest that necroptosis plays a key role upstream of coagulation in SIRS-related mortality. Furthermore, using a genetic lineage tracing mouse model (Tie2-Cre;Rosa26-tdT), we tracked endothelial cells (ECs) and verified that EC necroptosis is responsible for the vascular damage observed in TNF-α-treated mice. Importantly, Mlkl deletion in vascular ECs in mice had a similar protective effect against lethal SIRS by blocking EC necroptosis to protect the integrity of the endothelium. Collectively, our findings demonstrated that RIPK3–MLKL-dependent necroptosis disrupted vascular integrity, resulting in coagulopathy and multiorgan failure, eventually leading to mortality in SIRS patients. These results highlight the importance of targeting vascular EC necroptosis for the development of effective treatments for SIRS patients.

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Acknowledgements

We thank Dr. Xiaodong Wang (National Institute of Biological Sciences, Beijing, China) for providing Ripk3−/− mice. We also thank Zhonghui Weng (Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences) for animal studies, Lin Qiu (Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences) for flow cytometry technical support, and Yanqing Qin (Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences) for complete blood count analysis.

Funding

This work was supported by grants from the National Key Research and Development Program of China (2022YFA0807300), the National Natural Science Foundation of China (32270803, 32300630, 82272181, T2293734), the Shanghai Excellent Academic/Technical Leader Program (22XD1404500), the Shanghai Science and Technology Commission (23141902800), and the China Postdoctoral Science Foundation (2020M671261). We also acknowledge support from the Shanghai Municipal Science and Technology Major Project and the GuangCi Professorship Program of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine.

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Authors and Affiliations

  1. CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, PR China

    Xiaoxia Wu, Xiaoming Zhao, Yangjing Ou, Haiwei Zhang, Xiaoming Li, Xuanhui Wu, Lingxia Wang, Ming Li, Jianling Liu, Mingyan Xing, Han Liu, Yangyang Wang, Yangyang Xie, Hanwen Zhang, Hong Li, Yu Li & Haibing Zhang

  2. Department of Anesthesiology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China

    Fang Li

  3. Department of Microbiology and Immunology, Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China

    Yang Wang & Jing Wang

  4. Department of Anesthesiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China

    Yue Zhang, Yongchang Tan & Yan Luo

  5. State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, PR China

    Liming Sun

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  1. Xiaoxia Wu
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Contributions

Xiaoxia Wu and Haibing Zhang designed the study and analyzed the data; Xiaoxia Wu performed all the experiments with assistance from XMZ, FL, YW, YJO, HWZ, XML, XHW, LXW, ML, JLL, MYX, HL, YYW, YYX, and HWZ; YW and JW provided technical support for SD-IVM. YZ and YCT assisted with mouse genotyping and analysis. Yan Luo, LMS, HL, and Yu Li provided resources and intellectual input. Xiaoxia Wu and Haibing Zhang assembled the figure panels and wrote the paper. Haibing Zhang supervised the project.

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Correspondence to Haibing Zhang.

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All animal experiments were conducted in accordance with the guidelines of the Institutional Animal Care and Use Committee of the Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences.

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Wu, X., Zhao, X., Li, F. et al. MLKL-mediated endothelial necroptosis drives vascular damage and mortality in systemic inflammatory response syndrome. Cell Mol Immunol 21, 1309–1321 (2024). https://doi.org/10.1038/s41423-024-01217-y

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