Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles
Uremic toxins exert pathophysiological effects on cells and tissues, such as the generation of a pro-calcifying subtype of exosome-like extracellular vesicles (EVs) in vascular cells. Little is known about the effects of the toxins on the surface structure of EVs. Thus, we studied the effects of ure...
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MDPI AG
2023-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/18/14253 |
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| author | Christian Freise Andreas Zappe Norbert Löwa Jörg Schnorr Kevin Pagel Frank Wiekhorst Matthias Taupitz |
| author_facet | Christian Freise Andreas Zappe Norbert Löwa Jörg Schnorr Kevin Pagel Frank Wiekhorst Matthias Taupitz |
| author_sort | Christian Freise |
| collection | DOAJ |
| description | Uremic toxins exert pathophysiological effects on cells and tissues, such as the generation of a pro-calcifying subtype of exosome-like extracellular vesicles (EVs) in vascular cells. Little is known about the effects of the toxins on the surface structure of EVs. Thus, we studied the effects of uremic toxins on the abundance of sulfated glycosaminoglycans (GAGs) in EVs, and the implications for binding of ligands such as very small superparamagnetic iron oxide particles (VSOPs) which could be of relevance for radiological EV-imaging. Vascular cells were treated with the uremic toxins NaH<sub>2</sub>PO<sub>4</sub> and a mixture of urea and indoxyl sulfate. Uremia in rats was induced by adenine feeding. EVs were isolated from culture supernatants and plasma of rats. By proton T1-relaxometry, magnetic particle spectroscopy, and analysis of genes, proteins, and GAG-contents, we analyzed the roles of GAGs in the ligand binding of EVs. By influencing GAG-associated genes in host cells, uremic toxins induced higher GAG contents in EVs, particularly of sulfated chondroitin sulfate and heparan sulfate chains. EVs with high GAG content interacted stronger with VSOPs compared to control ones. This was confirmed by experiments with GAG-depleted EVs from genetically modified CHO cells and with uremic rat-derived EVs. Mechanistically, uremic toxin-induced PI3K/AKT-signaling and expression of the sulfate transporter SLC26A2 in host cells contributed to high GAG contents in EVs. In conclusion, uremic conditions induce enhanced GAG contents in EVs, which entails a stronger interaction with VSOPs. VSOPs might be suitable for radiological imaging of EVs rich in GAGs. |
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| last_indexed | 2024-03-10T22:39:15Z |
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| spelling | doaj.art-82f00ca30b7940cfb2a80cc53145c39c2023-11-19T11:10:03ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-09-0124181425310.3390/ijms241814253Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide ParticlesChristian Freise0Andreas Zappe1Norbert Löwa2Jörg Schnorr3Kevin Pagel4Frank Wiekhorst5Matthias Taupitz6Department of Radiology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Charitéplatz 1, 10117 Berlin, GermanyDepartment of Biology, Chemistry and Pharmacy, Freie Universität Berlin, Altensteinstraße 23A, 14195 Berlin, GermanyMetrology for Magnetic Nanoparticles Berlin, Physikalisch-Technische Bundesanstalt Berlin, Abbestr. 2, 10587 Berlin, GermanyDepartment of Radiology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Charitéplatz 1, 10117 Berlin, GermanyDepartment of Biology, Chemistry and Pharmacy, Freie Universität Berlin, Altensteinstraße 23A, 14195 Berlin, GermanyMetrology for Magnetic Nanoparticles Berlin, Physikalisch-Technische Bundesanstalt Berlin, Abbestr. 2, 10587 Berlin, GermanyDepartment of Radiology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Charitéplatz 1, 10117 Berlin, GermanyUremic toxins exert pathophysiological effects on cells and tissues, such as the generation of a pro-calcifying subtype of exosome-like extracellular vesicles (EVs) in vascular cells. Little is known about the effects of the toxins on the surface structure of EVs. Thus, we studied the effects of uremic toxins on the abundance of sulfated glycosaminoglycans (GAGs) in EVs, and the implications for binding of ligands such as very small superparamagnetic iron oxide particles (VSOPs) which could be of relevance for radiological EV-imaging. Vascular cells were treated with the uremic toxins NaH<sub>2</sub>PO<sub>4</sub> and a mixture of urea and indoxyl sulfate. Uremia in rats was induced by adenine feeding. EVs were isolated from culture supernatants and plasma of rats. By proton T1-relaxometry, magnetic particle spectroscopy, and analysis of genes, proteins, and GAG-contents, we analyzed the roles of GAGs in the ligand binding of EVs. By influencing GAG-associated genes in host cells, uremic toxins induced higher GAG contents in EVs, particularly of sulfated chondroitin sulfate and heparan sulfate chains. EVs with high GAG content interacted stronger with VSOPs compared to control ones. This was confirmed by experiments with GAG-depleted EVs from genetically modified CHO cells and with uremic rat-derived EVs. Mechanistically, uremic toxin-induced PI3K/AKT-signaling and expression of the sulfate transporter SLC26A2 in host cells contributed to high GAG contents in EVs. In conclusion, uremic conditions induce enhanced GAG contents in EVs, which entails a stronger interaction with VSOPs. VSOPs might be suitable for radiological imaging of EVs rich in GAGs.https://www.mdpi.com/1422-0067/24/18/14253extracellular vesiclesuremic toxinsVSOPglycosaminoglycansexosomes |
| spellingShingle | Christian Freise Andreas Zappe Norbert Löwa Jörg Schnorr Kevin Pagel Frank Wiekhorst Matthias Taupitz Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles International Journal of Molecular Sciences extracellular vesicles uremic toxins VSOP glycosaminoglycans exosomes |
| title | Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles |
| title_full | Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles |
| title_fullStr | Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles |
| title_full_unstemmed | Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles |
| title_short | Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles |
| title_sort | uremic toxin induced exosome like extracellular vesicles contain enhanced levels of sulfated glycosaminoglycans which facilitate the interaction with very small superparamagnetic iron oxide particles |
| topic | extracellular vesicles uremic toxins VSOP glycosaminoglycans exosomes |
| url | https://www.mdpi.com/1422-0067/24/18/14253 |
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