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Pseudomonas aeruginosa N-3-Oxododecanoyl Homoserine Lactone Disrupts Endothelial Integrity by Activating the Angiopoietin-Tie System

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Abstract

The activation of the angiopoietin (Angpt)-Tie system is linked to endothelial dysfunction during sepsis. Bacterial quorum-sensing molecules function as pathogen-associated molecular patterns. However, their impact on the endothelium and the Angpt-Tie system remains unclear. Therefore, this study investigated whether treatment with N-3-oxododecanoyl homoserine lactone (3OC12-HSL), a quorum-sensing molecule derived from Pseudomonas aeruginosa, impaired endothelial function in human umbilical vein endothelial cells. 3OC12-HSL treatment impaired tube formation even at sublethal concentrations, and immunocytochemistry analysis revealed that it seemed to reduce vascular endothelial-cadherin expression at the cell−cell interface. Upon assessing the mRNA expression patterns of genes associated with the Angpt-Tie axis, the expressions of Angpt2, Forkhead box protein O1, Tie1, and vascular endothelial growth factor 2 were found to be upregulated in the 3OC12-HSL-treated cells. Moreover, western blot analysis revealed that 3OC12-HSL treatment increased Angpt2 expression. A co-immunoprecipitation assay was conducted to assess the effect of 3OC12-HSL on the IQ motif containing GTPase activating protein 1 (IQGAP1) and Rac1 complex and the interaction between these proteins was consistently maintained regardless of 3OC12-HSL treatment. Next, recombinant human (rh)-Angpt1 was added to assess whether it modulated the effects of 3OC12-HSL treatment. rh-Angpt1 addition increased cellular viability, improved endothelial function, and reversed the overall patterns of mRNA and protein expression in endothelial cells treated with 3OC12-HSL. Additionally, it was related to the increased expression of phospho-Akt and the IQGAP1 and Rac1 complex. Collectively, our findings indicated that 3OC12-HSL from Pseudomonas aeruginosa can impair endothelial integrity via the activation of the Angpt-Tie axis, which appeared to be reversed by rh-Angpt1 treatment.

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References

  1. Singer, M., Deutschman, C. S., & Seymour, C. W., et al. (2016). The Third International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3). JAMA, 315(8), 801–810. https://doi.org/10.1001/jama.2016.0287.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Blanco, J., Muriel-Bombin, A., & Sagredo, V., et al. (2008). Incidence, organ dysfunction and mortality in severe sepsis: a Spanish multicentre study. Critical Care, 12(6), R158 https://doi.org/10.1186/cc7157.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Sakr, Y., Lobo, S. M., & Moreno, R. P., et al. (2012). Patterns and early evolution of organ failure in the intensive care unit and their relation to outcome. Critical Care, 16(6), R222. https://doi.org/10.1186/cc11868.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Ince, C., Mayeux, P. R., & Nguyen, T., et al. (2016). The endothelium in Sepsis. Shock, 45(3), 259–270. https://doi.org/10.1097/SHK.0000000000000473.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Lelubre, C., & Vincent, J. L. (2018). Mechanisms and treatment of organ failure in sepsis. Nature Reviews Nephrology, 14(7), 417–427. https://doi.org/10.1038/s41581-018-0005-7.

    Article  PubMed  Google Scholar 

  6. Koh, G. Y. (2013). Orchestral actions of angiopoietin-1 in vascular regeneration. Trends in Molecular Medicine, 19(1), 31–39. https://doi.org/10.1016/j.molmed.2012.10.010.

    Article  CAS  PubMed  Google Scholar 

  7. Kim, I., Kim, H. G., So, J. N., Kim, J. H., Kwak, H. J., & Koh, G. Y. (2000). Angiopoietin-1 regulates endothelial cell survival through the phosphatidylinositol 3’-Kinase/Akt signal transduction pathway. Circulation Research, 86(1), 24–29. https://doi.org/10.1161/01.res.86.1.24.

    Article  CAS  PubMed  Google Scholar 

  8. Kontos, C. D., Cha, E. H., York, J. D., & Peters, K. G. (2002). The endothelial receptor tyrosine kinase Tie1 activates phosphatidylinositol 3-kinase and Akt to inhibit apoptosis. Molecular and Cellular Biology, 22(6), 1704–1713. https://doi.org/10.1128/MCB.22.6.1704-1713.2002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Daly, C., Wong, V., & Burova, E., et al. (2004). Angiopoietin-1 modulates endothelial cell function and gene expression via the transcription factor FKHR (FOXO1). Genes Development, 18(9), 1060–1071. https://doi.org/10.1101/gad.1189704.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Gavard, J., Patel, V., & Gutkind, J. S. (2008). Angiopoietin-1 prevents VEGF-induced endothelial permeability by sequestering Src through mDia. Developmental Cells, 14(1), 25–36. https://doi.org/10.1016/j.devcel.2007.10.019.

    Article  CAS  Google Scholar 

  11. Maisonpierre, P. C., Suri, C., & Jones, P. F., et al. (1997). Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science, 277(5322), 55–60. https://doi.org/10.1126/science.277.5322.55.

    Article  CAS  PubMed  Google Scholar 

  12. Saharinen, P., Eklund, L., & Alitalo, K. (2017). Therapeutic targeting of the angiopoietin-TIE pathway. Nature Reviews Drug Discovery, 16(9), 635–661. https://doi.org/10.1038/nrd.2016.278.

    Article  CAS  PubMed  Google Scholar 

  13. Parikh, S. M., Mammoto, T., & Schultz, A., et al. (2006). Excess circulating angiopoietin-2 may contribute to pulmonary vascular leak in sepsis in humans. PLoS Medicine, 3(3), e46. https://doi.org/10.1371/journal.pmed.0030046.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. David, S., Mukherjee, A., & Ghosh, C. C., et al. (2012). Angiopoietin-2 may contribute to multiple organ dysfunction and death in sepsis. Critical Care Medicine, 40(11), 3034–3041. https://doi.org/10.1097/CCM.0b013e31825fdc31.

    Article  CAS  PubMed  Google Scholar 

  15. Lymperopoulou, K., Velissaris, D., & Kotsaki, A., et al. (2015). Angiopoietin-2 associations with the underlying infection and sepsis severity. Cytokine, 73(1), 163–168. https://doi.org/10.1016/j.cyto.2015.01.022.

    Article  CAS  PubMed  Google Scholar 

  16. Fang, Y., Li, C., Shao, R., Yu, H., Zhang, Q., & Zhao, L. (2015). Prognostic significance of the angiopoietin-2/angiopoietin-1 and angiopoietin-1/Tie-2 ratios for early sepsis in an emergency department. Critical Care, 19, 367. https://doi.org/10.1186/s13054-015-1075-6.

    Article  PubMed  PubMed Central  Google Scholar 

  17. Hughes, D. T., & Sperandio, V. (2008). Inter-kingdom signalling: communication between bacteria and their hosts. Nature Reviews Microbiology, 6(2), 111–120. https://doi.org/10.1038/nrmicro1836.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Whiteley, M., Diggle, S. P., & Greenberg, E. P. (2017). Progress in and promise of bacterial quorum sensing research. Nature, 551(7680), 313–320. https://doi.org/10.1038/nature24624.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Shiner, E. K., Terentyev, D., & Bryan, A., et al. (2006). Pseudomonas aeruginosa autoinducer modulates host cell responses through calcium signalling. Cellular Microbiology, 8(10), 1601–1610. https://doi.org/10.1111/j.1462-5822.2006.00734.x.

    Article  CAS  PubMed  Google Scholar 

  20. Skindersoe, M. E., Zeuthen, L. H., & Brix, S., et al. (2009). Pseudomonas aeruginosa quorum-sensing signal molecules interfere with dendritic cell-induced T-cell proliferation. FEMS Immunology and Medical Microbiology, 55(3), 335–345. https://doi.org/10.1111/j.1574-695X.2008.00533.x.

    Article  CAS  PubMed  Google Scholar 

  21. Vikstrom, E., Bui, L., Konradsson, P., & Magnusson, K. E. (2010). Role of calcium signalling and phosphorylations in disruption of the epithelial junctions by Pseudomonas aeruginosa quorum sensing molecule. European Journal of Cell Biology, 89(8), 584–597. https://doi.org/10.1016/j.ejcb.2010.03.002.

    Article  CAS  PubMed  Google Scholar 

  22. Karlsson, T., Turkina, M. V., Yakymenko, O., Magnusson, K. E., & Vikstrom, E. (2012). The Pseudomonas aeruginosa N-acylhomoserine lactone quorum sensing molecules target IQGAP1 and modulate epithelial cell migration. PLoS Pathogens, 8(10), e1002953. https://doi.org/10.1371/journal.ppat.1002953.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Schwarzer, C., Fu, Z., & Patanwala, M., et al. (2012). Pseudomonas aeruginosa biofilm-associated homoserine lactone C12 rapidly activates apoptosis in airway epithelia. Cellular Microbiology, 14(5), 698–709. https://doi.org/10.1111/j.1462-5822.2012.01753.x.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Glucksam-Galnoy, Y., Sananes, R., & Silberstein, N., et al. (2013). The bacterial quorum-sensing signal molecule N-3-oxo-dodecanoyl-L-homoserine lactone reciprocally modulates pro- and anti-inflammatory cytokines in activated macrophages. Journal of Immunology, 191(1), 337–344. https://doi.org/10.4049/jimmunol.1300368.

    Article  CAS  Google Scholar 

  25. Song, D., Meng, J., & Cheng, J., et al. (2019). Pseudomonas aeruginosa quorum-sensing metabolite induces host immune cell death through cell surface lipid domain dissolution. Nature Microbiology, 4(1), 97–111. https://doi.org/10.1038/s41564-018-0290-8.

    Article  CAS  PubMed  Google Scholar 

  26. Shin, J., Ahn, S. H., Kim, S. H., & Oh, D. J. (2021). N-3-oxododecanoyl homoserine lactone exacerbates endothelial cell death by inducing receptor-interacting protein kinase 1-dependent apoptosis. American Journal of Physiology-Cell Physiology, 321(4), C644–C653. https://doi.org/10.1152/ajpcell.00094.2021.

    Article  CAS  PubMed  Google Scholar 

  27. Brindle, N. P., Saharinen, P., & Alitalo, K. (2006). Signaling and functions of angiopoietin-1 in vascular protection. Circulation Research, 98(8), 1014–1023. https://doi.org/10.1161/01.RES.0000218275.54089.12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Smith, R. S., Harris, S. G., Phipps, R., & Iglewski, B. (2002). The Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxododecanoyl)homoserine lactone contributes to virulence and induces inflammation in vivo. Journal of Bacteriology, 184(4), 1132–1139. https://doi.org/10.1128/jb.184.4.1132-1139.2002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Vincent, J. L., Sakr, Y., & Sprung, C. L., et al. (2006). Sepsis in European intensive care units: results of the SOAP study. Critical Care Medicine, 34(2), 344–353. https://doi.org/10.1097/01.ccm.0000194725.48928.3a.

    Article  PubMed  Google Scholar 

  30. Blomquist, K. C., & Nix, D. E. (2021). A critical evaluation of newer beta-lactam antibiotics for treatment of Pseudomonas aeruginosa Infections. Annals of Pharmacotherapy, 55(8), 1010–1024. https://doi.org/10.1177/1060028020974003.

    Article  CAS  PubMed  Google Scholar 

  31. Menden, H., Welak, S., Cossette, S., Ramchandran, R., & Sampath, V. (2015). Lipopolysaccharide (LPS)-mediated angiopoietin-2-dependent autocrine angiogenesis is regulated by NADPH oxidase 2 (Nox2) in human pulmonary microvascular endothelial cells. Journal of Biological Chemistry, 290(9), 5449–5461. https://doi.org/10.1074/jbc.M114.600692.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Simons, M., Gordon, E., & Claesson-Welsh, L. (2016). Mechanisms and regulation of endothelial VEGF receptor signalling. Nature Reviews Molecular Cell Biology, 17(10), 611–625. https://doi.org/10.1038/nrm.2016.87.

    Article  CAS  PubMed  Google Scholar 

  33. Kawai, T., & Akira, S. (2007). Signaling to NF-kappaB by Toll-like receptors. Trends in Molecular Medicine, 13(11), 460–469. https://doi.org/10.1016/j.molmed.2007.09.002.

    Article  CAS  PubMed  Google Scholar 

  34. Ding, J., Song, D., Ye, X., & Liu, S. F. (2009). A pivotal role of endothelial-specific NF-kappaB signaling in the pathogenesis of septic shock and septic vascular dysfunction. Journal of Immunology, 183(6), 4031–4038. https://doi.org/10.4049/jimmunol.0900105.

    Article  CAS  Google Scholar 

  35. Kravchenko, V. V., Kaufmann, G. F., & Mathison, J. C., et al. (2006). N-(3-oxo-acyl)homoserine lactones signal cell activation through a mechanism distinct from the canonical pathogen-associated molecular pattern recognition receptor pathways. Journal of Biological Chemistry, 281(39), 28822–28830. https://doi.org/10.1074/jbc.M606613200.

    Article  CAS  PubMed  Google Scholar 

  36. Kravchenko, V. V., Kaufmann, G. F., & Mathison, J. C., et al. (2008). Modulation of gene expression via disruption of NF-kappaB signaling by a bacterial small molecule. Science, 321(5886), 259–263. https://doi.org/10.1126/science.1156499.

    Article  CAS  PubMed  Google Scholar 

  37. Vikstrom, E., Bui, L., Konradsson, P., & Magnusson, K. E. (2009). The junctional integrity of epithelial cells is modulated by Pseudomonas aeruginosa quorum sensing molecule through phosphorylation-dependent mechanisms. Experimental Cell Research, 315(2), 313–326. https://doi.org/10.1016/j.yexcr.2008.10.044.

    Article  CAS  PubMed  Google Scholar 

  38. Watanabe, T., Wang, S., & Noritake, J., et al. (2004). Interaction with IQGAP1 links APC to Rac1, Cdc42, and actin filaments during cell polarization and migration. Developmental Cells, 7(6), 871–883. https://doi.org/10.1016/j.devcel.2004.10.017.

    Article  CAS  Google Scholar 

  39. Noritake, J., Fukata, M., & Sato, K., et al. (2004). Positive role of IQGAP1, an effector of Rac1, in actin-meshwork formation at sites of cell-cell contact. Molecular Biology of the Cell, 15(3), 1065–1076. https://doi.org/10.1091/mbc.e03-08-0582.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Mataraza, J. M., Briggs, M. W., Li, Z., Entwistle, A., Ridley, A. J., & Sacks, D. B. (2003). IQGAP1 promotes cell motility and invasion. Journal of Biological Chemistry, 278(42), 41237–41245. https://doi.org/10.1074/jbc.M304838200.

    Article  CAS  PubMed  Google Scholar 

  41. David, S., Ghosh, C. C., Mukherjee, A., & Parikh, S. M. (2011). Angiopoietin-1 requires IQ domain GTPase-activating protein 1 to activate Rac1 and promote endothelial barrier defense. Arteriosclerosis, Thrombosis, and Vascular Biology, 31(11), 2643–2652. https://doi.org/10.1161/ATVBAHA.111.233189.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Korhonen, E. A., Lampinen, A., & Giri, H., et al. (2016). Tie1 controls angiopoietin function in vascular remodeling and inflammation. Journal of Clinical Investigation, 126(9), 3495–3510. https://doi.org/10.1172/JCI84923.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Cho, C. H., Kim, K. E., & Byun, J., et al. (2005). Long-term and sustained COMP-Ang1 induces long-lasting vascular enlargement and enhanced blood flow. Circulation Research, 97(1), 86–94. https://doi.org/10.1161/01.RES.0000174093.64855.a6.

    Article  CAS  PubMed  Google Scholar 

  44. David, S., Park, J. K., & Meurs, M., et al. (2011). Acute administration of recombinant Angiopoietin-1 ameliorates multiple-organ dysfunction syndrome and improves survival in murine sepsis. Cytokine, 55(2), 251–259. https://doi.org/10.1016/j.cyto.2011.04.005.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the National Research Foundation of Korea grant funded by the Korean government (MSIP) (No. 2020R1F1A1072498) and by departmental funding granted by Ma’am Sangchun Jung.

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  1. Department of Internal Medicine, Chung-Ang University College of Medicine, Seoul, South Korea

    Jungho Shin & Sun Hee Ahn

  2. Department of Internal Medicine, Myongji Hospital, Hanyang University College of Medicine, Goyang, South Korea

    Dong-Jin Oh

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Conceptualization, J.S. and D.J.O.; methodology, J.S., S.H.A., and D.J.O.; formal analysis and investigation, J.S. and S.H.A.; writing—original draft preparation, J.S. and D.J.O. All authors have read and agreed to the final version of the manuscript.

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Shin, J., Ahn, S.H. & Oh, DJ. Pseudomonas aeruginosa N-3-Oxododecanoyl Homoserine Lactone Disrupts Endothelial Integrity by Activating the Angiopoietin-Tie System. Cell Biochem Biophys (2024). https://doi.org/10.1007/s12013-024-01307-8

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