Exogenous loading of miRNAs into small extracellular vesicles.

Autor: de Abreu RC; CNC - Centro de Neurociências e Biologia Celular CIBB - Centro de Inovação em Biomedicina e Biotecnologia University of Coimbra Coimbra Portugal.; Faculty of Health, Medicine and Life Sciences CARIM School for Cardiovascular Diseases Maastricht University Maastricht The Netherlands.; Department of Molecular Genetics Faculty of Sciences and Engineering Maastricht University Maastricht The Netherlands., Ramos CV; Chemistry Department Faculty of Science and Technology Coimbra Chemistry Centre University of Coimbra Coimbra Portugal., Becher C; CNC - Centro de Neurociências e Biologia Celular CIBB - Centro de Inovação em Biomedicina e Biotecnologia University of Coimbra Coimbra Portugal., Lino M; CNC - Centro de Neurociências e Biologia Celular CIBB - Centro de Inovação em Biomedicina e Biotecnologia University of Coimbra Coimbra Portugal., Jesus C; CNC - Centro de Neurociências e Biologia Celular CIBB - Centro de Inovação em Biomedicina e Biotecnologia University of Coimbra Coimbra Portugal., da Costa Martins PA; Faculty of Health, Medicine and Life Sciences CARIM School for Cardiovascular Diseases Maastricht University Maastricht The Netherlands.; Department of Molecular Genetics Faculty of Sciences and Engineering Maastricht University Maastricht The Netherlands., Martins PAT; CNC - Centro de Neurociências e Biologia Celular CIBB - Centro de Inovação em Biomedicina e Biotecnologia University of Coimbra Coimbra Portugal.; Chemistry Department Faculty of Science and Technology Coimbra Chemistry Centre University of Coimbra Coimbra Portugal., Moreno MJ; Chemistry Department Faculty of Science and Technology Coimbra Chemistry Centre University of Coimbra Coimbra Portugal., Fernandes H; CNC - Centro de Neurociências e Biologia Celular CIBB - Centro de Inovação em Biomedicina e Biotecnologia University of Coimbra Coimbra Portugal.; Faculty of Medicine University of Coimbra Coimbra Portugal., Ferreira L; CNC - Centro de Neurociências e Biologia Celular CIBB - Centro de Inovação em Biomedicina e Biotecnologia University of Coimbra Coimbra Portugal.; Faculty of Medicine University of Coimbra Coimbra Portugal.
Jazyk: angličtina
Zdroj: Journal of extracellular vesicles [J Extracell Vesicles] 2021 Aug; Vol. 10 (10), pp. e12111. Date of Electronic Publication: 2021 Aug 02.
DOI: 10.1002/jev2.12111
Abstrakt: Small extracellular vesicles (sEVs), through their natural ability to interact with biological membranes and exploit endogenous processing pathways to convey biological information, are quintessential for the delivery of therapeutically relevant compounds, such as microRNAs (miRNAs) and proteins. Here, we used a fluorescently-labelled miRNA to quantify the efficiency of different methods to modulate the cargo of sEVs. Our results showed that, compared with electroporation, heat shock, permeation by a detergent-based compound (saponin) or cholesterol-modification of the miRNA, Exo-Fect was the most efficient method with > 50% transfection efficiency. Furthermore, qRT-PCR data showed that, compared with native sEVs, Exo-Fect modulation led to a > 1000-fold upregulation of the miRNA of interest. Importantly, this upregulation was observed for sEVs isolated from multiple sources. The modulated sEVs were able to delivery miR-155-5p into a reporter cell line, confirming the successful delivery of the miRNA to the target cell and, more importantly, its functionality. Finally, we showed that the membrane of Exo-Fect-loaded sEVs was altered compared with native sEVs and that enhanced the internalization of Exo-Fect-loaded sEVs within the target cells and decreased the interaction of those modulated sEVs with lysosomes.
(© 2021 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)
Databáze: MEDLINE
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