Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise
| Autor: | Eva-Maria Krämer-Albers, Elmo W. I. Neuberger, Perikles Simon, Katharina Mayr, Alexandra Brahmer, Barlo Hillen |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Adult
Male lcsh:QH426-470 Kinetics exosomes Extracellular vesicles Polymerase Chain Reaction Article 796 Athletic and outdoor sports and games 570 Life sciences cell-free DNA chemistry.chemical_compound Extracellular Vesicles Young Adult physical exercise Humans Exercise CD63 human plasma 796 Sport Chemistry Healthy Volunteers Cell biology lcsh:Genetics Long Interspersed Nucleotide Elements Human plasma Chromatography Gel Circulating DNA Female corona vesicular genomic DNA Cell-Free Nucleic Acids DNA CD81 extracellular DNA intraluminal 570 Biowissenschaften |
| Zdroj: | Genes Volume 12 Issue 4 Genes, Vol 12, Iss 522, p 522 (2021) |
| ISSN: | 2073-4425 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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