Cargando…

N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells

Mesenchymal stromal cell-derived extracellular vesicles (MSC-EV) are widely considered as a cell-free therapeutic alternative to MSC cell administration, due to their immunomodulatory and regenerative properties. However, the interaction mechanisms between EV and target cells are not fully understoo...

Descripción completa

Detalles Bibliográficos
Autores principales: Clos-Sansalvador, Marta, Garcia, Sergio G., Morón-Font, Miriam, Williams, Charles, Reichardt, Niels-Christian, Falcón-Pérez, Juan M., Bayes-Genis, Antoni, Roura, Santiago, Franquesa, Marcella, Monguió-Tortajada, Marta, Borràs, Francesc E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9455930/
https://www.ncbi.nlm.nih.gov/pubmed/36076936
http://dx.doi.org/10.3390/ijms23179539
_version_ 1784785686532980736
author Clos-Sansalvador, Marta
Garcia, Sergio G.
Morón-Font, Miriam
Williams, Charles
Reichardt, Niels-Christian
Falcón-Pérez, Juan M.
Bayes-Genis, Antoni
Roura, Santiago
Franquesa, Marcella
Monguió-Tortajada, Marta
Borràs, Francesc E.
author_facet Clos-Sansalvador, Marta
Garcia, Sergio G.
Morón-Font, Miriam
Williams, Charles
Reichardt, Niels-Christian
Falcón-Pérez, Juan M.
Bayes-Genis, Antoni
Roura, Santiago
Franquesa, Marcella
Monguió-Tortajada, Marta
Borràs, Francesc E.
author_sort Clos-Sansalvador, Marta
collection PubMed
description Mesenchymal stromal cell-derived extracellular vesicles (MSC-EV) are widely considered as a cell-free therapeutic alternative to MSC cell administration, due to their immunomodulatory and regenerative properties. However, the interaction mechanisms between EV and target cells are not fully understood. The surface glycans could be key players in EV–cell communication, being specific molecular recognition patterns that are still little explored. In this study, we focused on the role of N-glycosylation of MSC-EV as mediators of MSC-EV and endothelial cells’ interaction for subsequent EV uptake and the induction of cell migration and angiogenesis. For that, EV from immortalized Wharton’s Jelly MSC (iWJ-MSC-EV) were isolated by size exclusion chromatography (SEC) and treated with the glycosidase PNGase-F in order to remove wild-type N-glycans. Then, CFSE-labelled iWJ-MSC-EV were tested in the context of in vitro capture, agarose-spot migration and matrigel-based tube formation assays, using HUVEC. As a result, we found that the N-glycosylation in iWJ-MSC-EV is critical for interaction with HUVEC cells. iWJ-MSC-EV were captured by HUVEC, stimulating their tube-like formation ability and promoting their recruitment. Conversely, the removal of N-glycans through PNGase-F treatment reduced all of these functional activities induced by native iWJ-MSC-EV. Finally, comparative lectin arrays of iWJ-MSC-EV and PNGase-F-treated iWJ-MSC-EV found marked differences in the surface glycosylation pattern, particularly in N-acetylglucosamine, mannose, and fucose-binding lectins. Taken together, our results highlight the importance of N-glycans in MSC-EV to permit EV–cell interactions and associated functions.
format Online
Article
Text
id pubmed-9455930
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-94559302022-09-09 N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells Clos-Sansalvador, Marta Garcia, Sergio G. Morón-Font, Miriam Williams, Charles Reichardt, Niels-Christian Falcón-Pérez, Juan M. Bayes-Genis, Antoni Roura, Santiago Franquesa, Marcella Monguió-Tortajada, Marta Borràs, Francesc E. Int J Mol Sci Article Mesenchymal stromal cell-derived extracellular vesicles (MSC-EV) are widely considered as a cell-free therapeutic alternative to MSC cell administration, due to their immunomodulatory and regenerative properties. However, the interaction mechanisms between EV and target cells are not fully understood. The surface glycans could be key players in EV–cell communication, being specific molecular recognition patterns that are still little explored. In this study, we focused on the role of N-glycosylation of MSC-EV as mediators of MSC-EV and endothelial cells’ interaction for subsequent EV uptake and the induction of cell migration and angiogenesis. For that, EV from immortalized Wharton’s Jelly MSC (iWJ-MSC-EV) were isolated by size exclusion chromatography (SEC) and treated with the glycosidase PNGase-F in order to remove wild-type N-glycans. Then, CFSE-labelled iWJ-MSC-EV were tested in the context of in vitro capture, agarose-spot migration and matrigel-based tube formation assays, using HUVEC. As a result, we found that the N-glycosylation in iWJ-MSC-EV is critical for interaction with HUVEC cells. iWJ-MSC-EV were captured by HUVEC, stimulating their tube-like formation ability and promoting their recruitment. Conversely, the removal of N-glycans through PNGase-F treatment reduced all of these functional activities induced by native iWJ-MSC-EV. Finally, comparative lectin arrays of iWJ-MSC-EV and PNGase-F-treated iWJ-MSC-EV found marked differences in the surface glycosylation pattern, particularly in N-acetylglucosamine, mannose, and fucose-binding lectins. Taken together, our results highlight the importance of N-glycans in MSC-EV to permit EV–cell interactions and associated functions. MDPI 2022-08-23 /pmc/articles/PMC9455930/ /pubmed/36076936 http://dx.doi.org/10.3390/ijms23179539 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Clos-Sansalvador, Marta
Garcia, Sergio G.
Morón-Font, Miriam
Williams, Charles
Reichardt, Niels-Christian
Falcón-Pérez, Juan M.
Bayes-Genis, Antoni
Roura, Santiago
Franquesa, Marcella
Monguió-Tortajada, Marta
Borràs, Francesc E.
N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells
title N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells
title_full N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells
title_fullStr N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells
title_full_unstemmed N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells
title_short N-Glycans in Immortalized Mesenchymal Stromal Cell-Derived Extracellular Vesicles Are Critical for EV–Cell Interaction and Functional Activation of Endothelial Cells
title_sort n-glycans in immortalized mesenchymal stromal cell-derived extracellular vesicles are critical for ev–cell interaction and functional activation of endothelial cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9455930/
https://www.ncbi.nlm.nih.gov/pubmed/36076936
http://dx.doi.org/10.3390/ijms23179539
work_keys_str_mv AT clossansalvadormarta nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT garciasergiog nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT moronfontmiriam nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT williamscharles nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT reichardtnielschristian nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT falconperezjuanm nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT bayesgenisantoni nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT rourasantiago nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT franquesamarcella nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT monguiotortajadamarta nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells
AT borrasfrancesce nglycansinimmortalizedmesenchymalstromalcellderivedextracellularvesiclesarecriticalforevcellinteractionandfunctionalactivationofendothelialcells