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Lysosomal destabilization activates the NLRP3 inflammasome in human umbilical vein endothelial cells (HUVECs)

  • RESEARCH ARTICLE
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Journal of Cell Communication and Signaling Aims and scope

Abstract

Inflammation is a crucial component in the pathogenesis of many vascular diseases, such as atherosclerosis and diabetes. Inflammasomes are intracellular signalling complexes whose activation promotes inflammation. Nucleotide-binding domain and Leucine-rich repeat Receptor containing a Pyrin domain 3 (NLRP3) is a pattern recognition receptor (PRR) forming the best-known inflammasome. Disturbances in NLRP3 have been associated with multiple diseases. The purpose of this study was to explore the lysosomal destabilization-related NLRP3 inflammasome signaling pathway in human endothelial cells. In order to prime and activate NLRP3, human umbilical vein cells (HUVECs) were exposed to TNF-α and the lysosomal destructive agent Leusine-Leusine-O-Methylesther (Leu-Leu-OMe), respectively. A caspase-1 inhibitor was used to block caspase-1’s enzymatic function and an interleukin 1 receptor antagonist (IL-1RA) to prevent any possible secondary effects of IL-1β. Leu-Leu-OMe increased the expression of NLRP3, IL-1β, and IL-18 in HUVECs. Exposure to Leu-Leu-OMe significantly promoted the production of IL-6 and IL-8 in primed HUVECs; this effect was prevented by the pre-treatment of cells with an IL-1RA. Our results suggest that lysosomal destabilization activates the NLRP3 inflammasome pathway that promotes the production of IL-6 and IL-8 in an autocrine manner in HUVEC cells.

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Abbreviations

EC:

Endothelial cell

HUVEC:

Human umbilical vein endothelial cell

NLRP3:

Nucleotide-binding domain and leucine-rich repeat receptor containing a pyrin domain 3

IL:

Interleukin

IL-1RA:

Interleukin 1 receptor antagonist

PRR:

Pattern recognition receptor

Leu-Leu-OMe:

Leusine-Leusine-O-Methylesther

MVEC:

Microvascular endothelial cell

FBS:

Fetal bovine serum

TNF-α:

Tumor necrosis factor α

References

  • Abbate A, Kontos MC, Grizzard JD, Biondi-Zoccai GG, Van Tassell BW, Robati R et al (2010) Interleukin-1 blockade with anakinra to prevent adverse cardiac remodeling after acute myocardial infarction (virginia commonwealth university anakinra remodeling trial [VCU-ART] pilot study). Am J Cardiol 105:1371–1377.e1

    Article  CAS  PubMed  Google Scholar 

  • Aird WC (2007) Phenotypic heterogeneity of the endothelium: I. Structure, function, and mechanisms. Circ Res 100:158–173

    Article  CAS  PubMed  Google Scholar 

  • Bevilacqua MP, Pober JS, Wheeler ME, Cotran RS, Gimbrone MA Jr (1985) Interleukin-1 activation of vascular endothelium. Effects on procoagulant activity and leukocyte adhesion. Am J Pathol 121:394–403

    CAS  PubMed  PubMed Central  Google Scholar 

  • Chen Y, Li X, Boini KM, Pitzer AL, Gulbins E, Zhang Y et al (2015a) Endothelial Nlrp3 inflammasome activation associated with lysosomal destabilization during coronary arteritis. Biochim Biophys Acta 1853:396–408

    Article  CAS  PubMed  Google Scholar 

  • Chen Z, Wen L, Martin M, Hsu CY, Fang L, Lin FM et al (2015b) Oxidative stress activates endothelial innate immunity via sterol regulatory element binding protein 2 (SREBP2) transactivation of microRNA-92a. Circulation 131:805–814

    Article  CAS  PubMed  Google Scholar 

  • Chen Y, Wang L, Pitzer AL, Li X, Li PL, Zhang Y (2016) Contribution of redox-dependent activation of endothelial Nlrp3 inflammasomes to hyperglycemia-induced endothelial dysfunction. J Mol Med 94:1335–1347

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Duewell P, Kono H, Rayner KJ, Sirois CM, Vladimer G, Bauernfeind FG et al (2010) NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals. Nature 464:1357–1361

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ghasemi H, Ghazanfari T, Yaraee R, Faghihzadeh S, Hassan ZM (2011) Roles of IL-8 in ocular inflammations: a review. Ocul Immunol Inflamm 19:401–412

    Article  CAS  PubMed  Google Scholar 

  • Jaffe EA, Nachman RL, Becker CG, Minick CR (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52:2745–2756

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jo EK, Kim JK, Shin DM, Sasakawa C (2016) Molecular mechanisms regulating NLRP3 inflammasome activation. Cell Mol Immunol 13:148–159

    Article  CAS  PubMed  Google Scholar 

  • Kaarniranta K, Sinha D, Blasiak J, Kauppinen A, Vereb Z, Salminen A et al (2013) Autophagy and heterophagy dysregulation leads to retinal pigment epithelium dysfunction and development of age-related macular degeneration. Autophagy 9:973–984

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kauppinen A, Paterno JJ, Blasiak J, Salminen A, Kaarniranta K (2016) Inflammation and its role in age-related macular degeneration. Cell Mol Life Sci 73:1765–1786

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Larsen CM, Faulenbach M, Vaag A, Volund A, Ehses JA, Seifert B et al (2007) Interleukin-1-receptor antagonist in type 2 diabetes mellitus. N Engl J Med 356:1517–1526

    Article  CAS  PubMed  Google Scholar 

  • Lee HM, Kim JJ, Kim HJ, Shong M, Ku BJ, Jo EK (2013) Upregulated NLRP3 inflammasome activation in patients with type 2 diabetes. Diabetes 62:194–204

    Article  CAS  PubMed  Google Scholar 

  • Loukovaara S, Kinnunen K, Piippo N, Hytti M, Kaarniranta K, Kauppinen A (2017) NLRP3 inflammasome activation is associated with proliferative diabetic retinopathy. Acta Ophthalmol In press. doi:10.1111/aos.13427

  • Menu P, Pellegrin M, Aubert JF, Bouzourene K, Tardivel A, Mazzolai L et al (2011) Atherosclerosis in ApoE-deficient mice progresses independently of the NLRP3 inflammasome. Cell Death Dis 2:e137

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Minciullo PL, Catalano A, Mandraffino G, Casciaro M, Crucitti A, Maltese G et al (2016) Inflammaging and anti-inflammaging: the role of cytokines in extreme longevity. Arch Immunol Ther Exp 64:111–126

    Article  CAS  Google Scholar 

  • Nathan C (2002) Points of control in inflammation. Nature 420:846–852

    Article  CAS  Google Scholar 

  • Nixon RA (2013) The role of autophagy in neurodegenerative disease. Nat Med 19:983–997

    Article  CAS  PubMed  Google Scholar 

  • O’Connor W Jr, Harton JA, Zhu X, Linhoff MW, Ting JP (2003) Cutting edge: CIAS1/Cryopyrin/PYPAF1/NALP3/ CATERPILLER 1.1 is an inducible inflammatory mediator with NF-κB suppressive properties. J Immunol 171:6329–6333

    Article  PubMed  Google Scholar 

  • Piippo N, Korkmaz A, Hytti M, Kinnunen K, Salminen A, Atalay M et al (2014) Decline in cellular clearance systems induces inflammasome signaling in human ARPE-19 cells. Biochim Biophys Acta 1843:3038–3046

    Article  CAS  PubMed  Google Scholar 

  • Salminen A, Kaarniranta K, Kauppinen A (2012) Inflammaging: disturbed interplay between autophagy and inflammasomes. Aging 4:166–175

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sandanger O, Ranheim T, Vinge LE, Bliksoen M, Alfsnes K, Finsen AV et al (2013) The NLRP3 inflammasome is up-regulated in cardiac fibroblasts and mediates myocardial ischaemia-reperfusion injury. Cardiovasc Res 99:164–174

    Article  CAS  PubMed  Google Scholar 

  • Tseng WA, Thein T, Kinnunen K, Lashkari K, Gregory MS, D'Amore PA et al (2013) NLRP3 inflammasome activation in retinal pigment epithelial cells by lysosomal destabilization: implications for age-related macular degeneration. Invest Ophthalmol Vis Sci 54:110–120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Van Tassell BW, Arena RA, Toldo S, Mezzaroma E, Azam T, Seropian IM et al (2012) Enhanced interleukin-1 activity contributes to exercise intolerance in patients with systolic heart failure. PLoS One 7:e33438

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang L, Chen Y, Li X, Zhang Y, Gulbins E, Zhang Y (2016) Enhancement of endothelial permeability by free fatty acid through lysosomal cathepsin B-mediated Nlrp3 inflammasome activation. Oncotarget 7:73229–73241

    PubMed  PubMed Central  Google Scholar 

  • Webster BR, Scott I, Traba J, Han K, Sack MN (2014) Regulation of autophagy and mitophagy by nutrient availability and acetylation. Biochim Biophys Acta 1841:525–534

  • Xiao H, Lu M, Lin TY, Chen Z, Chen G, Wang WC et al (2013) Sterol regulatory element binding protein 2 activation of NLRP3 inflammasome in endothelium mediates hemodynamic-induced atherosclerosis susceptibility. Circulation 128:632–642

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We warmly acknowledge Prof. Seppo Ylä-Herttuala for providing cells for our study, Mrs. Anne Seppänen for technical support, Dr. Ewen MacDonald for the language revision, and Res. Dir. Emeritus Antero Salminen for the valuable collaboration, discussions, and critical review of the manuscript. This study was financially supported by Päivikki ja Sakari Sohlberg Foundation, Finnish Eye Foundation, Finnish Cultural Foundation (Central and North-Savo Regional Fund), Orion-Farmos Research Foundation, the Alfred Kordelin Foundation, Emil Aaltonen Foundation, Mary and Georg C Ehrnrooth Foundation, Kuopio University Hospital (VTR funding), the Academy of Finland (Health Research Council projects AK297267, AK307341, and KK5503743), and the Helsinki University Central Hospital (HUCH) Clinical Research Grant TYH2016230.

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Authors and Affiliations

  1. Department of Ophthalmology, Kuopio University Hospital, P.O. Box 100, FI-70029, Kuopio, Finland

    K. Kinnunen & K. Kaarniranta

  2. Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, P.O. Box 1627, FI-70211, Kuopio, Finland

    K. Kinnunen, N. Piippo & K. Kaarniranta

  3. School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, P.O.B. 1627, FI-70211, Kuopio, Finland

    N. Piippo, M. Hytti & A. Kauppinen

  4. Unit of Vitreoretinal Surgery, Department of Ophthalmology, University of Helsinki and Helsinki University Hospital, FI-00029, Helsinki, Finland

    S. Loukovaara

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  1. K. Kinnunen
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  2. N. Piippo
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  3. S. Loukovaara
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  4. M. Hytti
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  5. K. Kaarniranta
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  6. A. Kauppinen
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Correspondence to K. Kinnunen.

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Kinnunen, K., Piippo, N., Loukovaara, S. et al. Lysosomal destabilization activates the NLRP3 inflammasome in human umbilical vein endothelial cells (HUVECs). J. Cell Commun. Signal. 11, 275–279 (2017). https://doi.org/10.1007/s12079-017-0396-4

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  • DOI: https://doi.org/10.1007/s12079-017-0396-4

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