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Neutrophil extracellular traps induced by IL-1β promote endothelial dysfunction and aggravate limb ischemia

Hypertension Research (2024)Cite this article

Abstract

Vascular inflammation and endothelial dysfunction contribute to vascular diseases. While neutrophil extracellular traps (NETs) participate in some vascular pathologies, their roles in lower limb ischemia remain poorly defined. This study investigated the functional significance of NETs in vascular inflammation and remodeling associated with limb ischemia. Single-cell RNA sequencing (scRNA-seq) and flow cytometry revealed neutrophil activation and upregulated NETs formation in human limb ischemia, with immunofluorescence confirming IL-1β-induced release of NETs for vascular inflammation. Endothelial cell activation was examined via scRNA-seq and western blotting, indicating enhanced proliferation, expression of adhesion molecules (VCAM-1, ICAM-1), inflammatory cytokines (IL-1β, IL-6) and decreased expression of VE-cadherin, that could be mediated by NETs to exacerbate endothelial inflammation. Mechanistically, NETs altered endothelial cell function via increased pSTAT1/STAT1 signaling. Vascular inflammation and subsequent ischemia were alleviated in vivo by NETosis or IL-1β inhibition in ischemic mice. IL-1β-NETs induce endothelial activation and inflammation in limb ischemia by stimulating STAT1 signaling. Targeting NETs may thus represent a novel therapeutic strategy for inflammatory vascular diseases associated with limb ischemia.

Graphical abstract of NETs regulation of the development of vascular inflammation in lower limb ischemia via pSTAT1/STAT1 signaling pathway.

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The data underlying this article will be shared on reasonable request to the corresponding author.

References

  1. Libby P. Inflammation in atherosclerosis. Nature. 2002;420:868–74.

    Article  CAS  PubMed  Google Scholar 

  2. Annex BH, Cooke JP. New directions in therapeutic angiogenesis and arteriogenesis in peripheral arterial disease. Circ Res. 2021;128:1944–57.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Xu S, Ilyas I, Little PJ, Li H, Kamato D, Zheng X, et al. Endothelial dysfunction in atherosclerotic cardiovascular diseases and beyond: from mechanism to pharmacotherapies. Pharmacol Rev. 2021;73:924–67.

    Article  CAS  PubMed  Google Scholar 

  4. Loukogeorgakis SP, Williams R, Panagiotidou AT, Kolvekar SK, Donald A, Cole TJ, et al. Transient limb ischemia induces remote preconditioning and remote postconditioning in humans by a K(ATP)-channel dependent mechanism. Circulation. 2007;116:1386–95.

    Article  CAS  PubMed  Google Scholar 

  5. Medzhitov R. The spectrum of inflammatory responses. Science. 2021;374:1070–5.

    Article  CAS  PubMed  Google Scholar 

  6. Lutgens E, Atzler D, Doring Y, Duchene J, Steffens S, Weber C. Immunotherapy for cardiovascular disease. Eur Heart J. 2019;40:3937–46.

    Article  CAS  PubMed  Google Scholar 

  7. Meher AK, Spinosa M, Davis JP, Pope N, Laubach VE, Su G, et al. Novel role of IL (Interleukin)-1beta in neutrophil extracellular trap formation and abdominal aortic aneurysms. Arteriosclerosis Thrombosis Vascular Biol. 2018;38:843–53.

    Article  CAS  Google Scholar 

  8. Liew PX, Kubes P. The neutrophil’s role during health and disease. Physiol Rev. 2019;99:1223–48.

    Article  CAS  PubMed  Google Scholar 

  9. Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, et al. Neutrophil extracellular traps kill bacteria. Science. 2004;303:1532–5.

    Article  CAS  PubMed  Google Scholar 

  10. Papayannopoulos V. Neutrophil extracellular traps in immunity and disease. Nat Rev Immunol. 2018;18:134–47.

    Article  CAS  PubMed  Google Scholar 

  11. Scheiermann C, Kunisaki Y, Frenette PS. Circadian control of the immune system. Nat Rev Immunol. 2013;13:190–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Man K, Loudon A, Chawla A. Immunity around the clock. Science. 2016;354:999–1003.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Adrover JM, Del Fresno C, Crainiciuc G, Cuartero MI, Casanova-Acebes M, Weiss LA, et al. A neutrophil timer coordinates immune defense and vascular protection. Immunity. 2019;50:390–402.e10.

    Article  CAS  PubMed  Google Scholar 

  14. Castanheira FVS, Kubes P. Neutrophils and NETs in modulating acute and chronic inflammation. Blood. 2019;133:2178–85.

    Article  CAS  PubMed  Google Scholar 

  15. Doring Y, Libby P, Soehnlein O. Neutrophil extracellular traps participate in cardiovascular diseases: recent experimental and clinical insights. Circ Res. 2020;126:1228–41.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Ridker PM. Anticytokine agents: targeting interleukin signaling pathways for the treatment of atherothrombosis. Circ Res. 2019;124:437–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Song P, Rudan D, Zhu Y, Fowkes FJI, Rahimi K, Fowkes FGR, et al. Global, regional, and national prevalence and risk factors for peripheral artery disease in 2015: an updated systematic review and analysis. Lancet Glob Health. 2019;7:e1020–e30.

    Article  PubMed  Google Scholar 

  18. Ridker PM. How common is residual inflammatory risk? Circ Res. 2017;120:617–9.

    Article  CAS  PubMed  Google Scholar 

  19. Unamuno X, Gomez-Ambrosi J, Ramirez B, Rodriguez A, Becerril S, Valenti V, et al. NLRP3 inflammasome blockade reduces adipose tissue inflammation and extracellular matrix remodeling. Cell Mol Immunol. 2021;18:1045–57.

    Article  CAS  PubMed  Google Scholar 

  20. Aarreberg LD, Esser-Nobis K, Driscoll C, Shuvarikov A, Roby JA, Gale M Jr. Interleukin-1beta Induces mtDNA Release to Activate Innate Immune Signaling via cGAS-STING. Mol Cell. 2019;74:801–15.e6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Jones SA, Jenkins BJ. Recent insights into targeting the IL-6 cytokine family in inflammatory diseases and cancer. Nat Rev Immunol. 2018;18:773–89.

    Article  CAS  PubMed  Google Scholar 

  22. Nathan C. Neutrophils and immunity: challenges and opportunities. Nat Rev Immunol. 2006;6:173–82.

    Article  CAS  PubMed  Google Scholar 

  23. Bhattacharya M, Berends ETM, Chan R, Schwab E, Roy S, Sen CK, et al. Staphylococcus aureus biofilms release leukocidins to elicit extracellular trap formation and evade neutrophil-mediated killing. Proc Natl Acad Sci USA. 2018;115:7416–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Etulain J, Martinod K, Wong SL, Cifuni SM, Schattner M, Wagner DD. P-selectin promotes neutrophil extracellular trap formation in mice. Blood. 2015;126:242–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Jung CJ, Yeh CY, Hsu RB, Lee CM, Shun CT, Chia JS. Endocarditis pathogen promotes vegetation formation by inducing intravascular neutrophil extracellular traps through activated platelets. Circulation. 2015;131:571–81.

    Article  CAS  PubMed  Google Scholar 

  26. Brinkmann V, Zychlinsky A. Neutrophil extracellular traps: is immunity the second function of chromatin? J Cell Biol. 2012;198:773–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Wang L, Zhou X, Yin Y, Mai Y, Wang D, Zhang X. Hyperglycemia induces neutrophil extracellular traps formation through an NADPH oxidase-dependent pathway in diabetic retinopathy. Front Immunol. 2018;9:3076.

    Article  CAS  PubMed  Google Scholar 

  28. Yousefi S, Mihalache C, Kozlowski E, Schmid I, Simon HU. Viable neutrophils release mitochondrial DNA to form neutrophil extracellular traps. Cell Death Differ. 2009;16:1438–44.

    Article  CAS  PubMed  Google Scholar 

  29. Wigerblad G, Kaplan MJ. Neutrophil extracellular traps in systemic autoimmune and autoinflammatory diseases. Nat Rev Immunol. 2023;23:274–88.

    Article  CAS  PubMed  Google Scholar 

  30. Xiao Y, Cong M, Li J, He D, Wu Q, Tian P, et al. Cathepsin C promotes breast cancer lung metastasis by modulating neutrophil infiltration and neutrophil extracellular trap formation. Cancer Cell. 2021;39:423–37.e7.

    Article  CAS  PubMed  Google Scholar 

  31. Tian Z, Zhang Y, Zheng Z, Zhang M, Zhang T, Jin J, et al. Gut microbiome dysbiosis contributes to abdominal aortic aneurysm by promoting neutrophil extracellular trap formation. Cell Host Microbe. 2022;30:1450–63.e8.

    Article  CAS  PubMed  Google Scholar 

  32. Kumar SV, Kulkarni OP, Mulay SR, Darisipudi MN, Romoli S, Thomasova D, et al. Neutrophil extracellular trap-related extracellular histones cause vascular necrosis in severe GN. J Am Soc Nephrol. 2015;26:2399–413.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Furchgott RF. The 1996 Albert Lasker Medical Research Awards. The discovery of endothelium-derived relaxing factor and its importance in the identification of nitric oxide. JAMA. 1996;276:1186–8.

    Article  CAS  PubMed  Google Scholar 

  34. Murad F. Shattuck Lecture. Nitric oxide and cyclic GMP in cell signaling and drug development. N Engl J Med. 2006;355:2003–11.

    Article  CAS  PubMed  Google Scholar 

  35. Al Rifai M, DeFilippis AP, McEvoy JW, Hall ME, Acien AN, Jones MR, et al. The relationship between smoking intensity and subclinical cardiovascular injury: The Multi-Ethnic Study of Atherosclerosis (MESA). Atherosclerosis. 2017;258:119–30.

    Article  CAS  PubMed  Google Scholar 

  36. DiStasi MR, Ley K. Opening the flood-gates: how neutrophil-endothelial interactions regulate permeability. Trends Immunol. 2009;30:547–56.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Hu G, Vogel SM, Schwartz DE, Malik AB, Minshall RD. Intercellular adhesion molecule-1-dependent neutrophil adhesion to endothelial cells induces caveolae-mediated pulmonary vascular hyperpermeability. Circ Res. 2008;102:e120–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Ridker PM, Everett BM, Thuren T, MacFadyen JG, Chang WH, Ballantyne C, et al. Antiinflammatory therapy with canakinumab for atherosclerotic disease. N Engl J Med. 2017;377:1119–31.

    Article  CAS  PubMed  Google Scholar 

  39. Wu B, Xu MM, Fan C, Feng CL, Lu QK, Lu HM, et al. STING inhibitor ameliorates LPS-induced ALI by preventing vascular endothelial cells-mediated immune cells chemotaxis and adhesion. Acta Pharmacol Sin. 2022;43:2055–66.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We appreciate for the technical support by the Core Facilities, Central Laboratory, the First Affiliated Hospital, Zhejiang University School of Medicine.

Funding

This work was supported by the National Natural Science Foundation of China (82370496, 81700420, 32300945) and the Zhejiang Provincial Natural Science Foundation of China under Grant No. LQ23H020006.

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Authors and Affiliations

  1. Department of Vascular Surgery, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

    Shigang Lin, Yujian Hu, Lirui Huang, Yangyan He & Hongkun Zhang

  2. Department of Cardiology, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China

    Pengwei Zhu & Liujun Jiang

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Contributions

HZ and YH conceived and designed the study. SL, PZ and LJ performed the experiments. YH and LH established the model of hind limb ischemic. PZ and LJ analyzed the data. SL wrote the manuscript.

Corresponding authors

Correspondence to Yangyan He or Hongkun Zhang.

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The authors declare no competing interests.

Ethics approval

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Research Ethics Committee of the First Afliated Hospital of Zhejiang University School of Medicine (approval No. 2020/98).

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Informed consent was obtained from all individual participants included in the study.

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Lin, S., Zhu, P., Jiang, L. et al. Neutrophil extracellular traps induced by IL-1β promote endothelial dysfunction and aggravate limb ischemia. Hypertens Res (2024). https://doi.org/10.1038/s41440-024-01661-3

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