3D visualization of extracellular vesicle uptake by endothelial cells

• Autorzy: Durak-Kozica M. , Baster Z. , Kubat K. , Stepien E.
• Języki publikacji: EN

Abstrakty

EN

Background: Extracellular vesicles are small vesicles that contain cytoplasmic and membrane components from their paternal cells. They enter target cells through uptake to transfer their biological cargo. In this study, we investigated the process of endothelial EV internalization and created a 3D visualization of their intracellular distribution. Methods and results: Two immortalized endothelial cell lines that express h-TERT (human telomerase) were used for EV release: microvascular TIME and macrovascular HUVEC. EVs were isolated from the cell culture medium via differential centrifugation and used for the uptake experiments. The size distribution of the EVs was measured using TRPS technology on a qNano instrument. Internalization of EVs was observed using a Zeiss LSM 710 confocal laser microscope after staining of the EVs with PKH26. EVs were observed intracellularly and distributed in the perinuclear region of the target cells. The distribution patterns were similar in both cell lines. Conclusion: The perinuclear localization of the internalized EVs shows their biological stability after their uptake to the endothelial cells. The 3D visualization allows the determination of a more accurate location of EVs relative to the donor cell nucleus.

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2018
• Tom: 23
• Opis fizyczny: p.1-9,fig.,ref.

Twórcy

autor

Durak-Kozica M.

• Department of Medical Physics, Institute of Physics, Jagiellonian University, Lojasiewicza 11, Kraków, Poland

autor

Baster Z.

• Department of Medical Physics, Institute of Physics, Jagiellonian University, Lojasiewicza 11, Kraków, Poland

autor

Kubat K.

• Department of Medical Physics, Institute of Physics, Jagiellonian University, Lojasiewicza 11, Kraków, Poland

autor

Stepien E.

• Department of Medical Physics, Institute of Physics, Jagiellonian University, Lojasiewicza 11, Kraków, Poland

Bibliografia

• 1. Choi DS, Kim DK, Kim YK, Ys G. Proteomics, transcriptomics and lipidomics of exosomes and Ectosomes. Proteomics. 2013;13:1554–71.
• 2. Raposo G, Stoorvogel W. Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol. 2013;200:373–83.
• 3. Kim DK, Lee J, Kim SR, Choi DS, Yoon YJ, Kim JH, et al. EVpedia: a community web portal for extracellular vesicles research. Bioinformatics. 2015;31:933–9.
• 4. Gajos K, Kamińska K, Awsiuk K, Bajor A, Gruszczyński K, Pawlak A, et al. Immobilization and detection of platelet-derived extracellular vesicles on functionalized silicon substrate: cytometric and spectrometric approach. Anal Bioanal Chem. 2017;409:1109–19.
• 5. Montecalvo A, Larregina AT, Shufesky WJ, Stolz DB, Sullivan ML, Karlsson JM, et al. Mechanism of transfer of functional microRNAs between mouse dendritic cells via exosomes. Blood. 2012;119:756–66.
• 6. Nazarenko I, Rana S, Baumann A, McAlear J, Hellwig A, Trendelenburg M, et al. Cell Surface tetraspanin Tspan8 contributes to molecular pathways of exosome-induced Endothelial cell activation. Cancer Res. 2010;70:1668–78.
• 7. Deregibus MC, Cantaluppi V, Calogero R, Lo Iacono M, Tetta C, Biancone L, et al. Endothelial progenitor cell derived microvesicles activate an angiogenic program in endothelial cells by a horizontal transfer of mRNA. Blood. 2007;110:2440–8.
• 8. Kirchhausen T. Clathrin. Annu Rev Biochem. 2000;69:699–727.
• 9. Wang LH, Rothberg KG, Anderson RG. Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol. 1993;123:1107–17.
• 10. Doherty GJ, McMahon HT. Mechanisms of endocytosis. Annu Rev Biochem. 2009;78:857–902.
• 11. Ahram M, Sameni M, Qiu RG, Linebaugh B, Kirn D, Sloane BF. Rac1-induced endocytosis is associated with intracellular proteolysis during migration through a three-dimensional matrix. Exp Cell Res. 2000;260:292–303.
• 12. Ridley AJ. Rho GTPases and actin dynamics in membrane protrusions and vesicle trafficking. Trends Cell Biol. 2006;16:522–9.
• 13. Swanson JA. Shaping cups into phagosomes and macropinosomes. Nat Rev Mol Cell Biol. 2008;9:639–49.
• 14. Barre’ SC, Blanc L, Bette-Bobillo P, Andre’ S, Mamoun R, Gabius HJ, et al. Galectin-5 is bound onto the surface of rat reticulocyte exosomes and modulates vesicle uptake by macrophages. Blood. 2010;115:696–705.
• 15. Nanbo A, Kawanishi E, Yoshida R, Yoshiyama H. Exosomes derived from Epstein-Barr virus-infected cells are internalized via caveola-dependent endocytosis and promote phenotypic modulation in target cells. J Virol. 2013;87:10334–47.
• 16. Menck K, Klemm F, Gross JC, Pukrop T, Wenzel D, Binder C. Induction and transport of Wnt 5a during macrophageinduced malignant invasion is mediated by two types of extracellular vesicles. Oncotarget. 2013;4:2057–66.
• 17. Nabi IR, Le PU. Caveolae/raft-dependent endocytosis. J Cell Biol. 2003;161:673–7.
• 18. Simons K, Ehehalt R. Cholesterol, lipid rafts, and disease. J Clin Invest. 2002;110:597–603.
• 19. Teissier E, EI P´c. Lipids as modulators of membrane fusion mediated by viral fusion proteins. Eur Biophys J. 2007;36: 887–99.
• 20. Christianson HC, Svensson KJ, van Kuppevelt TH, Li JP, Belting M. Cancer cell exosomes depend on cell-surface heparan sulfate proteoglycans for their internalization and functional activity. Proc Natl Acad Sci U S A. 2013;110:17380–5.
• 21. Hemler ME. Tetraspanin functions and associated microdomains. Nat Rev Mol Cell Biol. 2005;6:801–11.
• 22. Zoller M. Tetraspanins: push and pull in suppressing and promoting metastasis. Nat Rev Cancer. 2009;9:40–55.
• 23. Levy S, Shoham T. The tetraspanin web modulates immunesignalling complexes. Nat Rev Immunol. 2005;5:136–48.
• 24. Vjugina U, Evans JP. New insights into the molecular basis of mammalian sperm-egg membrane interactions. Front Biosci. 2008;13:462–76.
• 25. Morelli AE, Larregina AT, Shufesky WJ, Sullivan ML, Stolz DB, Papworth GD, et al. Endocytosis, intracellular sorting, and processing of exosomes by dendritic cells. Blood. 2004;104:3257–66.
• 26. Aplin AE, Howe A, Alahari SK, Juliano RL. Signal transduction and signal modulation by cell adhesion receptors: the role of integrins, cadherins, immunoglobulin-cell adhesion molecules. and selectins Pharmacol Rev. 1998;50:197–263.
• 27. Voyta JC, Via DP, Butterfield CE, Zetter RB. Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J Cell Biol. 1984;99:2034–40.
• 28. Costa Verdera H, Gitz-Francois JJ, Schiffelers RM, Vader P. Cellular uptake of extracellular vesicles is mediated by clathrinindependent endocytosis and macropinocytosis. J Control Release. 2017;266:100–8.
• 29. Sáez T, Salsoso R, Leiva A, Toledo F, de Vos P, Faas M, et al. Human umbilical vein endothelium-derived exosomes play a role in foetoplacental endothelial dysfunction in gestational diabetes mellitus. Biochim Biophys Acta. 2018;1864:499–508.
• 30. Lee JK, Park SR, Jung BK, Jeon YK, Lee YS, Kim MK, et al. Exosomes derived from mesenchymal stem cells suppress angiogenesis by down-regulating VEGF expression in breast cancer cells. PLoS One. 2013;8:84256.
• 31. Terrisse AD, Puech N, Allart S, Gourdy P, Xuereb JM, Payrastre B, et al. Internalization of microparticles by endothelial cells promotes platelet/endothelial cell interaction under flow. J Thromb Haemost. 2010;8:2810–9.
• 32. Kamińska A, Enguita FJ, Stępień EŁ. Lactadherin: An unappreciated haemostasis regulator and potential therapeutic agent. Vasc Pharmacol. 2018;101:21–8.
• 33. Mantel PY, Hjelmqvist D, Walch M, Kharoubi-Hess S, Nilsson S, Ravel D, et al. Infected erythrocyte-derived extracellular vesicles alter vascular function via regulatory Ago2-miRNA complexes in malaria. Nat Commun. 2016. https://doi.org/10.1038/ncomms12727.
• 34. Lombardo G, Dentelli P, Togliatto G, Rosso A, Gili M, Gallo S, et al. Activated Stat5 trafficking Via Endothelial Cell-derived Extracellular Vesicles Controls IL-3 Pro-angiogenic Paracrine Action. Sci Rep. 2016. https://doi.org/10.1038/srep25689.
• 35. Tian T, Wang Y, Wang H, Zhu Z, Xiao Z. Visualizing of the cellular uptake and intracellular trafficking of exosomes by live-cell microscopy. J Cell Biochem. 2010;111:488–96.
• 36. Tian T, Zhu YL, Hu FH, Wang YY, Huang NP, Xiao ZD. Dynamics of exosome internalization and trafficking. J Cell Physiol. 2012:228, 1487–1295.
• 37. Escrevente C, Keller S, Altevogt P, Costa J. Interaction and uptake of exosomes by ovarian cancer cells. BMC Cancer. 2011;11:108.
• 38. Stefani C, Lacy-Hulbert A, Skillman T. Confocal VR: Immersive Visualization for Confocal Microscopy. J Mol Biol. 2018. https://doi.org/10.1016/j.jmb.2018.06.035.
• 39. Servier. Servier Medical Art licensed under CC BY 3.0. 2018. https://smart.servier.com/category/general-items/equipment/ laboratory-equipment. Accessed 22 Jul 2018.

Identyfikatory

DOI

10.1186/s11658-018-0123-z