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Autoren: Neuberger, Elmo W. I.
Hillen, Barlo
Mayr, Katharina
Simon, Perikles
Krämer-Albers, Eva Maria
Brahmer, Alexandra
Titel: Kinetics and topology of DNA associated with circulating extracellular vesicles released during exercise
Online-Publikationsdatum: 12-Nov-2021
Sprache des Dokuments: Englisch
Zusammenfassung/Abstract: Although it is widely accepted that cancer-derived extracellular vesicles (EVs) carry DNA cargo, the association of cell-free circulating DNA (cfDNA) and EVs in plasma of healthy humans remains elusive. Using a physiological exercise model, where EVs and cfDNA are synchronously released, we aimed to characterize the kinetics and localization of DNA associated with EVs. EVs were separated from human plasma using size exclusion chromatography or immuno-affinity capture for CD9+, CD63+, and CD81+ EVs. DNA was quantified with an ultra-sensitive qPCR assay targeting repetitive LINE elements, with or without DNase digestion. This model shows that a minute part of circulating cell-free DNA is associated with EVs. During rest and following exercise, only 0.12% of the total cfDNA occurs in association with CD9+/CD63+/CD81+EVs. DNase digestion experiments indicate that the largest part of EV associated DNA is sensitive to DNase digestion and only ~20% are protected within the lumen of the separated EVs. A single bout of running or cycling exercise increases the levels of EVs, cfDNA, and EV-associated DNA. While EV surface DNA is increasing, DNAse-resistant DNA remains at resting levels, indicating that EVs released during exercise (ExerVs) do not contain DNA. Consequently, DNA is largely associated with the outer surface of circulating EVs. ExerVs recruit cfDNA to their corona, but do not carry DNA in their lumen.
DDC-Sachgruppe: 570 Biowissenschaften
570 Life sciences
796 Sport
796 Athletic and outdoor sports and games
Veröffentlichende Institution: Johannes Gutenberg-Universität Mainz
Organisationseinheit: FB 02 Sozialwiss., Medien u. Sport
Veröffentlichungsort: Mainz
DOI: http://doi.org/10.25358/openscience-6493
Version: Published version
Publikationstyp: Zeitschriftenaufsatz
Nutzungsrechte: CC BY
Informationen zu den Nutzungsrechten: https://creativecommons.org/licenses/by/4.0/
Zeitschrift: Genes
12
4
Seitenzahl oder Artikelnummer: 522
Verlag: MDPI
Verlagsort: Basel
Erscheinungsdatum: 2021
ISSN: 2073-4425
URL der Originalveröffentlichung: https://doi.org/10.3390/genes12040522
DOI der Originalveröffentlichung: 10.3390/genes12040522
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