Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise
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...
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MDPI AG
2021-04-01
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| Series: | Genes |
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| Online Access: | https://www.mdpi.com/2073-4425/12/4/522 |
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| author | Elmo W. I. Neuberger Barlo Hillen Katharina Mayr Perikles Simon Eva-Maria Krämer-Albers Alexandra Brahmer |
| author_facet | Elmo W. I. Neuberger Barlo Hillen Katharina Mayr Perikles Simon Eva-Maria Krämer-Albers Alexandra Brahmer |
| author_sort | Elmo W. I. Neuberger |
| collection | DOAJ |
| description | 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<sup>+</sup>, CD63<sup>+</sup>, and CD81<sup>+</sup> 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<sup>+</sup>/CD63<sup>+</sup>/CD81<sup>+</sup>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. |
| first_indexed | 2024-03-10T12:38:29Z |
| format | Article |
| id | doaj.art-3d7754d6d40d4f4699fc1b067c45e8de |
| institution | Directory Open Access Journal |
| issn | 2073-4425 |
| language | English |
| last_indexed | 2024-03-10T12:38:29Z |
| publishDate | 2021-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Genes |
| spelling | doaj.art-3d7754d6d40d4f4699fc1b067c45e8de2023-11-21T14:04:20ZengMDPI AGGenes2073-44252021-04-0112452210.3390/genes12040522Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during ExerciseElmo W. I. Neuberger0Barlo Hillen1Katharina Mayr2Perikles Simon3Eva-Maria Krämer-Albers4Alexandra Brahmer5Department of Sports Medicine, Rehabilitation and Disease Prevention, Johannes Gutenberg University Mainz, 55099 Mainz, GermanyDepartment of Sports Medicine, Rehabilitation and Disease Prevention, Johannes Gutenberg University Mainz, 55099 Mainz, GermanyInstitute of Developmental Biology and Neurobiology, Extracellular Vesicles Research Group, Johannes Gutenberg University Mainz, 55099 Mainz, GermanyDepartment of Sports Medicine, Rehabilitation and Disease Prevention, Johannes Gutenberg University Mainz, 55099 Mainz, GermanyInstitute of Developmental Biology and Neurobiology, Extracellular Vesicles Research Group, Johannes Gutenberg University Mainz, 55099 Mainz, GermanyDepartment of Sports Medicine, Rehabilitation and Disease Prevention, Johannes Gutenberg University Mainz, 55099 Mainz, GermanyAlthough 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<sup>+</sup>, CD63<sup>+</sup>, and CD81<sup>+</sup> 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<sup>+</sup>/CD63<sup>+</sup>/CD81<sup>+</sup>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.https://www.mdpi.com/2073-4425/12/4/522extracellular vesiclesexosomescell-free DNAextracellular DNAcoronaintraluminal |
| spellingShingle | Elmo W. I. Neuberger Barlo Hillen Katharina Mayr Perikles Simon Eva-Maria Krämer-Albers Alexandra Brahmer Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise Genes extracellular vesicles exosomes cell-free DNA extracellular DNA corona intraluminal |
| title | Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise |
| title_full | Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise |
| title_fullStr | Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise |
| title_full_unstemmed | Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise |
| title_short | Kinetics and Topology of DNA Associated with Circulating Extracellular Vesicles Released during Exercise |
| title_sort | kinetics and topology of dna associated with circulating extracellular vesicles released during exercise |
| topic | extracellular vesicles exosomes cell-free DNA extracellular DNA corona intraluminal |
| url | https://www.mdpi.com/2073-4425/12/4/522 |
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