nach oben

European Journal of Applied Physiology

Erschienen in:

01.11.2015 | Original Article

Release of bulk cell free DNA during physical exercise occurs independent of extracellular vesicles

verfasst von: Susanne Helmig, Carsten Frühbeis, Eva-Maria Krämer-Albers, Perikles Simon, Suzan Tug

Erschienen in: European Journal of Applied Physiology | Ausgabe 11/2015

Einloggen, um Zugang zu erhalten

Abstract

Purpose

Strenuous exercise induces a rapid and transient elevation of cell free DNA (cfDNA) concentration in blood plasma. The detection of cfDNA in the presence of plasma nucleases could indicate an association of cfDNA with protective vesicular structures. Several cell types release extracellular vesicles (EVs), including exosomes and shedding microvesicles, which are known to mediate the exchange of proteins and nucleic acids (largely RNA) between cells. Here, we assessed whether EVs play a role in the exercise-dependent release of cfDNA in blood plasma.

Methods

Venous blood collected from healthy volunteers before and after incremental treadmill exercise was separated into vesicular (EV) and soluble fractions. Nuclear and mitochondrial DNA content in plasma supernatants and EV fractions was determined by quantitative real-time PCR (qPCR).

Results

We show that the majority of cfDNA is located in the plasma supernatants. Only minute amounts of DNA were observed in the EV-associated fractions including microvesicles and exosomes. Nuclear and mitochondrial DNA species differ in terms of their quantities in the several plasma fractions.

Conclusions

Our results indicate that cfDNA liberated in response to acute physical exercise is not released by vesicular means and circulates in a soluble form in blood plasma which could indicate different biological functions exerted by cfDNA and EVs. The different nature of DNA species in plasma has major implications for the preparation of plasma and other bodily fluids prior to analysis.

Nur mit Berechtigung zugänglich

Aatonen M, Gronholm M, Siljander PR (2012) Platelet-derived microvesicles: multitalented participants in intercellular communication. Semin Thromb Hemost 38:102–113. doi:10.1055/s-0031-1300956 CrossRefPubMed

Adeva-Andany M, Lopez-Ojen M, Funcasta-Calderon R, Ameneiros-Rodriguez E, Donapetry-Garcia C, Vila-Altesor M, Rodriguez-Seijas J (2014) Comprehensive review on lactate metabolism in human health. Mitochondrion 17C:76–100. doi:10.1016/j.mito.2014.05.007 CrossRef

Atamaniuk J, Vidotto C, Tschan H, Bachl N, Stuhlmeier KM, Muller MM (2004) Increased concentrations of cell-free plasma DNA after exhaustive exercise. Clin Chem 50:1668–1670. doi:10.1373/clinchem.2004.034553 CrossRefPubMed

Atamaniuk J, Stuhlmeier KM, Vidotto C, Tschan H, Dossenbach-Glaninger A, Mueller MM (2008) Effects of ultra-marathon on circulating DNA and mRNA expression of pro- and anti-apoptotic genes in mononuclear cells. Eur J Appl Physiol 104:711–717. doi:10.1007/s00421-008-0827-2 CrossRefPubMed

Atamaniuk J, Vidotto C, Kinzlbauer M, Bachl N, Tiran B, Tschan H (2010) Cell-free plasma DNA and purine nucleotide degradation markers following weightlifting exercise. Eur J Appl Physiol 110:695–701. doi:10.1007/s00421-010-1532-5 CrossRefPubMed

Balaj L, Lessard R, Dai L, Cho YJ, Pomeroy SL, Breakefield XO, Skog J (2011) Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences. Nature Commun 2:180. doi:10.1038/ncomms1180 CrossRef

Beiter T, Fragasso A, Hudemann J, Niess AM, Simon P (2011) Short-term treadmill running as a model for studying cell-free DNA kinetics in vivo. Clin Chem 57:633–636. doi:10.1373/clinchem.2010.158030 CrossRefPubMed

Beiter T, Fragasso A, Hudemann J, Schild M, Steinacker J, Mooren FC, Niess AM (2014) Neutrophils release extracellular DNA traps in response to exercise. J Appl Physiol 117:325–333. doi:10.1152/japplphysiol.00173.2014 CrossRefPubMed

Bellingham SA, Guo BB, Coleman BM, Hill AF (2012) Exosomes: vehicles for the transfer of toxic proteins associated with neurodegenerative diseases? Front Physiol 3:124. doi:10.3389/fphys.2012.00124 PubMedCentralCrossRefPubMed

Blagosklonny MV (2000) Cell death beyond apoptosis Leukemia 14:1502–1508PubMed

Bobrie A, Colombo M, Raposo G, Thery C (2011) Exosome secretion: molecular mechanisms and roles in immune responses. Traffic 12:1659–1668. doi:10.1111/j.1600-0854.2011.01225.x CrossRefPubMed

Breitbach S, Tug S, Simon P (2012) Circulating cell-free DNA: an up-coming molecular marker in exercise physiology. Sports Med 42:565–586. doi:10.2165/11631380-000000000-00000 CrossRefPubMed

Breitbach S, Sterzing B, Magallanes C, Tug S, Simon P (2014) Direct measurement of cell-free DNA from serially collected capillary plasma during incremental exercise. J Appl Physiol 117:119–130. doi:10.1152/japplphysiol.00002.2014 CrossRefPubMed

Budnik LT, Kloth S, Baur X, Preisser AM, Schwarzenbach H (2013) Circulating mitochondrial DNA as biomarker linking environmental chemical exposure to early preclinical lesions elevation of mtDNA in human serum after exposure to carcinogenic halo-alkane-based pesticides. PLoS One 8:e64413. doi:10.1371/journal.pone.0064413 PubMedCentralCrossRefPubMed

Ceci R et al (2013) Oxidative stress responses to a graded maximal exercise test in older adults following explosive-type resistance training. Redox Biol 2C:65–72. doi:10.1016/j.redox.2013.12.004 PubMed

Chaar V et al (2011) Effect of strenuous physical exercise on circulating cell-derived microparticles. Clin Hemorheol Microcirc 47:15–25. doi:10.3233/CH-2010-1361 PubMed

Chiu RW, Chan LY, Lam NY, Tsui NB, Ng EK, Rainer TH, Lo YM (2003) Quantitative analysis of circulating mitochondrial DNA in plasma. Clin Chem 49:719–726CrossRefPubMed

Dickhuth HH, Yin L, Niess A, Rocker K, Mayer F, Heitkamp HC, Horstmann T (1999) Ventilatory, lactate-derived and catecholamine thresholds during incremental treadmill running: relationship and reproducibility. Int J Sports Med 20:122–127. doi:10.1055/s-2007-971105 PubMed

El Messaoudi S, Rolet F, Mouliere F, Thierry AR (2013) Circulating cell free DNA: preanalytical considerations. Clin Chim Acta 424:222–230. doi:10.1016/j.cca.2013.05.022 CrossRefPubMed

Fatouros IG et al (2010) Time of sampling is crucial for measurement of cell-free plasma DNA following acute aseptic inflammation induced by exercise. Clin Biochem 43:1368–1370. doi:10.1016/j.clinbiochem.2010.08.020 CrossRefPubMed

Fruhbeis C, Frohlich D, Kuo WP, Kramer-Albers EM (2013) Extracellular vesicles as mediators of neuron-glia communication. Front Cell Neurosci 7:182. doi:10.3389/fncel.2013.00182 PubMedCentralCrossRefPubMed

Frühbeis C, Helmig S, Tug S, Simon P, Krämer-Albers EM (2015) Physical exercise induces rapid release of small extracellular vesicles into the circulation. J Extracell Vesicles 4:28239. doi:10.3402/jev.v4.28239 CrossRefPubMed

Galeazzi M, Morozzi G, Piccini M, Chen J, Bellisai F, Fineschi S, Marcolongo R (2003) Dosage and characterization of circulating DNA: present usage and possible applications in systemic autoimmune disorders. Autoimmun Rev 2:50–55CrossRefPubMed

Gould SJ, Raposo G (2013) As we wait: coping with an imperfect nomenclature for extracellular vesicles. J Extracell Vesicles. doi:10.3402/jev.v2i0.20389 PubMedCentralPubMed

Harding CV, Heuser JE, Stahl PD (2013) Exosomes: looking back three decades and into the future. J Cell Biol 200:367–371. doi:10.1083/jcb.201212113 PubMedCentralCrossRefPubMed

Hergenreider E et al (2012) Atheroprotective communication between endothelial cells and smooth muscle cells through miRNAs. Nat Cell Biol 14:249–256. doi:10.1038/ncb2441 CrossRefPubMed

Holdenrieder S, Stieber P (2009) Clinical use of circulating nucleosomes. Crit Rev Clin Lab Sci 46:1–24. doi:10.1080/10408360802485875 CrossRefPubMed

Horstman LL, Jy W, Minagar A, Bidot CJ, Jimenez JJ, Alexander JS, Ahn YS (2007) Cell-derived microparticles and exosomes in neuroinflammatory disorders. Int Rev Neurobiol 79:227–268. doi:10.1016/S0074-7742(07)79010-4 CrossRefPubMed

Huang X et al (2013) Characterization of human plasma-derived exosomal RNAs by deep sequencing. BMC Genom 14:319. doi:10.1186/1471-2164-14-319 CrossRef

Jahr S, Hentze H, Englisch S, Hardt D, Fackelmayer FO, Hesch RD, Knippers R (2001) DNA fragments in the blood plasma of cancer patients: quantitations and evidence for their origin from apoptotic and necrotic cells. Cancer Res 61:1659–1665PubMed

Kahlert C et al (2014) Identification of double-stranded genomic DNA spanning all chromosomes with mutated KRAS and p53 DNA in the serum exosomes of patients with pancreatic cancer. J Biol Chem 289:3869–3875. doi:10.1074/jbc.C113.532267 PubMedCentralCrossRefPubMed

Kalra H et al (2012) Vesiclepedia: a compendium for extracellular vesicles with continuous community annotation. PLoS Biol 10:e1001450. doi:10.1371/journal.pbio.1001450 PubMedCentralCrossRefPubMed

Kowal J, Tkach M, Thery C (2014) Biogenesis and secretion of exosomes. Curr Opin Cell Biol 29C:116–125. doi:10.1016/j.ceb.2014.05.004 CrossRef

Krämer-Albers EM et al (2007) Oligodendrocytes secrete exosomes containing major myelin and stress-protective proteins: trophic support for axons? Proteomics Clin Appl 1:1446–1461CrossRefPubMed

Lazaro-Ibanez E, Sanz-Garcia A, Visakorpi T, Escobedo-Lucea C, Siljander P, Ayuso-Sacido A, Yliperttula M (2014) Different gDNA content in the subpopulations of prostate cancer extracellular vesicles: apoptotic bodies, microvesicles, and exosomes. Prostate. doi:10.1002/pros.22853 PubMedCentralPubMed

Lee TH, Chennakrishnaiah S, Audemard E, Montermini L, Meehan B, Rak J (2014) Oncogenic ras-driven cancer cell vesiculation leads to emission of double-stranded DNA capable of interacting with target cells. Biochem Biophys Res Commun. doi:10.1016/j.bbrc.2014.07.109

Luga V et al (2012) Exosomes mediate stromal mobilization of autocrine Wnt-PCP signaling in breast cancer cell migration. Cell 151:1542–1556. doi:10.1016/j.cell.2012.11.024 CrossRefPubMed

Lui YY, Chik KW, Chiu RW, Ho CY, Lam CW, Lo YM (2002) Predominant hematopoietic origin of cell-free DNA in plasma and serum after sex-mismatched bone marrow transplantation. Clin Chem 48:421–427PubMed

Miranda KC et al (2010) Nucleic acids within urinary exosomes/microvesicles are potential biomarkers for renal disease. Kidney Int 78:191–199. doi:10.1038/ki.2010.106 PubMedCentralCrossRefPubMed

Nagata S, Hanayama R, Kawane K (2010) Autoimmunity and the clearance of dead cells. Cell 140:619–630. doi:10.1016/j.cell.2010.02.014 CrossRefPubMed

Peinado H et al (2012) Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nat Med 18:883–891. doi:10.1038/nm.2753 PubMedCentralCrossRefPubMed

Pilsczek FH et al (2010) A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus. J Immunol 185:7413–7425. doi:10.4049/jimmunol.1000675 CrossRefPubMed

Pisetsky DS (2009) Microparticles as biomarkers in autoimmunity: from dust bin to center stage. Arthritis Res Ther 11:135. doi:10.1186/ar2856 PubMedCentralCrossRefPubMed

Raposo G, Stoorvogel W (2013) Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol 200:373–383. doi:10.1083/jcb.201211138 PubMedCentralCrossRefPubMed

Revenfeld AL, Baek R, Nielsen MH, Stensballe A, Varming K, Jorgensen M (2014) Diagnostic and prognostic potential of extracellular vesicles in peripheral blood. Clin Ther 36:830–846. doi:10.1016/j.clinthera.2014.05.008 CrossRefPubMed

Rhead B et al (2010) The UCSC Genome Browser database: update. Nucleic Acids Res 38:D613–D619. doi:10.1093/nar/gkp939 PubMedCentralCrossRefPubMed

Robbins PD, Morelli AE (2014) Regulation of immune responses by extracellular vesicles. Nat Rev Immunol 14:195–208. doi:10.1038/nri3622 PubMedCentralCrossRefPubMed

Schwarzenbach H, Hoon DS, Pantel K (2011) Cell-free nucleic acids as biomarkers in cancer patients. Nat Rev Cancer 11:426–437. doi:10.1038/nrc3066 CrossRefPubMed

Simons M, Raposo G (2009) Exosomes–vesicular carriers for intercellular communication. Curr Opin Cell Biol 21:575–581. doi:10.1016/j.ceb.2009.03.007 CrossRefPubMed

Swarup V, Rajeswari MR (2007) Circulating (cell-free) nucleic acids–a promising, non-invasive tool for early detection of several human diseases. FEBS Lett 581:795–799. doi:10.1016/j.febslet.2007.01.051 CrossRefPubMed

Thakur BK et al (2014) Double-stranded DNA in exosomes: a novel biomarker in cancer detection. Cell Res 24:766–769. doi:10.1038/cr.2014.44 PubMedCentralCrossRefPubMed

Thery C (2011) Exosomes: secreted vesicles and intercellular communications. F1000 Biol Rep 3:15. doi:10.3410/B3-15 PubMedCentralCrossRefPubMed

Thery C, Ostrowski M, Segura E (2009) Membrane vesicles as conveyors of immune responses. Nat Rev Immunol 9:581–593. doi:10.1038/nri2567 CrossRefPubMed

Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO (2007) Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9:654–659. doi:10.1038/ncb1596 CrossRefPubMed

Walsh NP et al (2011) Position statement. Part one: Immune function and exercise. Exerc Immunol Rev 17:6–63PubMed

Yi J et al (2014) Increased plasma cell-free DNA level during HTNV infection: correlation with disease severity and virus load. Viruses 6:2723–2734. doi:10.3390/v6072723 PubMedCentralCrossRefPubMed

Ziegler A, Zangemeister-Wittke U, Stahel RA (2002) Circulating DNA: a new diagnostic gold mine? Cancer Treat Rev 28:255–271CrossRefPubMed

Titel: Release of bulk cell free DNA during physical exercise occurs independent of extracellular vesicles
verfasst von: Susanne Helmig
Carsten Frühbeis
Eva-Maria Krämer-Albers
Perikles Simon
Suzan Tug
Publikationsdatum: 01.11.2015
Verlag: Springer Berlin Heidelberg
Erschienen in: European Journal of Applied Physiology / Ausgabe 11/2015
Print ISSN: 1439-6319
Elektronische ISSN: 1439-6327
DOI: https://doi.org/10.1007/s00421-015-3207-8

Neu im Fachgebiet Arbeitsmedizin

Elterliches Belastungserleben, Unaufmerksamkeits‑/Hyperaktivitätssymptome und elternberichtete ADHS bei Kindern und Jugendlichen: Ergebnisse aus der KiGGS-Studie

Open Access ADHS Leitthema

Die Aufmerksamkeitsdefizit‑/Hyperaktivitätsstörung (ADHS) ist eine der häufigsten psychischen Störungen im Kindes- und Jugendalter [ 1 ]. In Deutschland beträgt die Prävalenz einer elternberichteten ADHS-Diagnose bei Kindern und Jugendlichen 4,4 % …

Substanzkonsum und Nutzung von sozialen Medien, Computerspielen und Glücksspielen unter Auszubildenden an beruflichen Schulen

Open Access Leitthema

Die Begrenzung von Schäden durch Substanzkonsum und andere abhängige Verhaltensweisen von Jugendlichen und jungen Erwachsenen ist ein wichtiges Anliegen der öffentlichen Gesundheit. Der Übergang von der Adoleszenz zum jungen Erwachsenenalter ist …

Berufsbelastung und Stressbewältigung von weiblichen und männlichen Auszubildenden

Leitthema

In der Öffentlichkeit wird die berufliche Ausbildung oftmals unter ökonomischen Gesichtspunkten diskutiert: Mit den geburtenstarken Jahrgängen gehen erfahrene Fachkräfte in Rente und von nachfolgenden Generationen rücken zu wenige Arbeitskräfte …

Rauschtrinken in der frühen Adoleszenz

COVID-19 Leitthema

Alkohol ist in Deutschland die mit Abstand am häufigsten konsumierte psychoaktive Substanz. Mehr als 2 Drittel aller Erwachsenen im Alter von 18 bis 64 Jahren (70,5 %) hat 2021 in den letzten 30 Tagen Alkohol konsumiert [ 1 ]. Von diesen …

Springer Medizin

Abstract

Purpose

Methods

Results

Conclusions

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

Weitere Artikel der Ausgabe 11/2015

Effects of breathing frequency on the heart rate deceleration capacity and heart rate acceleration capacity

New Zealand blackcurrant extract improves cycling performance and fat oxidation in cyclists

Acute response to cluster sets in trained and untrained men

Haemodynamics of aerobic and resistance blood flow restriction exercise in young and older adults

The effect of transcranial direct current stimulation of the motor cortex on exercise-induced pain

Local and remote thermoregulatory changes affect NIRS measurement in forearm muscles

Neu im Fachgebiet Arbeitsmedizin

Elterliches Belastungserleben, Unaufmerksamkeits‑/Hyperaktivitätssymptome und elternberichtete ADHS bei Kindern und Jugendlichen: Ergebnisse aus der KiGGS-Studie

Substanzkonsum und Nutzung von sozialen Medien, Computerspielen und Glücksspielen unter Auszubildenden an beruflichen Schulen

Berufsbelastung und Stressbewältigung von weiblichen und männlichen Auszubildenden

Rauschtrinken in der frühen Adoleszenz