Release of Mitochondrial and Nuclear DNA During On-Pump Heart Surgery: Kinetics and Relation to Extracellular Vesicles

  • Anton Baysa
  • Anton Fedorov
  • Kirill Kondratov
  • Arno Ruusalepp
  • Sarkis Minasian
  • Michael Galagudza
  • Maxim Popov
  • Dmitry Kurapeev
  • Alexey Yakovlev
  • Guro Valen
  • Anna Kostareva
  • Jarle Vaage
  • Kåre-Olav StensløkkenEmail author
Original Article


During heart surgery with cardiopulmonary bypass (CPB), the release of mitochondrial (mtDNA) and nuclear DNA (nDNA) and their association to extracellular vesicles were investigated. In patients undergoing elective coronary artery bypass grafting (CABG, n = 12), blood was sampled before, during, and after surgery from peripheral artery, pulmonary artery, and the coronary sinus. Plasma was separated in three fractions: microvesicles, exosomes, and supernatant. mtDNA and nDNA were measured by qPCR. mtDNA and nDNA levels increased after start of surgery, but before CPB, and increased further during CPB. mtDNA copy number was about 1000-fold higher than nDNA. mtDNA was predominantly localized to the vesicular fractions in plasma, whereas nDNA was predominantly in the supernatant. The amount of free mtDNA increased after surgery. There was no net release or disappearance of DNAs across the pulmonary, systemic, or coronary circulation. Extracellular DNAs, in particular mtDNA, may be important contributors to the whole-body inflammation during CPB.


Extracellular DNA Cardiac surgery Exosomes Microvesicles 



We gratefully acknowledge Torun Flatebø for technical support with qPCR and Maksim Puzanov for assistance in sample collection. Low-voltage scanning electron microscopy was performed using equipment of the Interdisciplinary Resource Center for Nanotechnology of Saint Petersburg State University.


This study was funded by the Norwegian Research Council (#214557), Gjensidige stiftelsen, Anders Jahres fund, Russian Science Foundation (#17-75-30052), and the University of Oslo.

Compliance with Ethical Standards

The studies were performed in agreement with the ethical standards stated in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. The studies were approved by the Regional Ethics Committee in Oslo, Norway; Tartu University Hospital, Estonia; or the Institutional Ethics Committee at Almazov National Medical Research Centre, Saint Petersburg, Russia. Written, informed consent was obtained from all patients.

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12265_2018_9848_MOESM1_ESM.docx (16 kb)
ESM 1 (DOCX 15 kb)


  1. 1.
    Atamaniuk, J., Ruzicka, K., Stuhlmeier, K. M., Karimi, A., Eigner, M., & Mueller, M. M. (2006). Cell-free plasma DNA: a marker for apoptosis during hemodialysis. Clinical Chemistry, 52, 523–526. Scholar
  2. 2.
    Bakhle, Y. S., & Vane, J. R. (1974). Pharmacokinetic function of the pulmonary circulation. Physiological Reviews, 54, 1007–1045. Scholar
  3. 3.
    Bhagirath, V. C., Dwivedi, D. J., & Liaw, P. C. (2015). Comparison of the proinflammatory and procoagulant properties of nuclear, mitochondrial, and bacterial DNA. Shock, 44, 265–271. Scholar
  4. 4.
    Bliksoen, M., Mariero, L. H., Ohm, I. K., Haugen, F., Yndestad, A., Solheim, S., Seljeflot, I., Ranheim, T., Andersen, G. O., Aukrust, P., Valen, G., & Vinge, L. E. (2012). Increased circulating mitochondrial DNA after myocardial infarction. International Journal of Cardiology, 158, 132–134. Scholar
  5. 5.
    Bliksoen, M., Mariero, L. H., Torp, M. K., Baysa, A., Ytrehus, K., Haugen, F., Seljeflot, I., Vaage, J., Valen, G., & Stenslokken, K. O. (2016). Extracellular mtDNA activates NF-kappaB via toll-like receptor 9 and induces cell death in cardiomyocytes. Basic Research in Cardiology, 111, 42. Scholar
  6. 6.
    Burger, G., Gray, M. W., & Lang, B. F. (2003). Mitochondrial genomes: anything goes. Trends in Genetics, 19, 709–716. Scholar
  7. 7.
    Chiu, R. W., Chan, L. Y., Lam, N. Y., Tsui, N. B., Ng, E. K., Rainer, T. H., & Lo, Y. M. (2003). Quantitative analysis of circulating mitochondrial DNA in plasma. Clinical Chemistry, 49, 719–726.CrossRefGoogle Scholar
  8. 8.
    Edmunds, L. H., Jr. (1998). Inflammatory response to cardiopulmonary bypass. The Annals of Thoracic Surgery, 66, S12–S16; discussion S25-18. Scholar
  9. 9.
    Ellefsen, S., Stenslokken, K. O., Sandvik, G. K., Kristensen, T. A., & Nilsson, G. E. (2008). Improved normalization of real-time reverse transcriptase polymerase chain reaction data using an external RNA control. Analytical Biochemistry, 376, 83–93. Scholar
  10. 10.
    Frangogiannis, N. G. (2014). The inflammatory response in myocardial injury, repair, and remodelling. Nature Reviews. Cardiology, 11, 255–265. Scholar
  11. 11.
    Gray, M. W., Burger, G., & Lang, B. F. (1999). Mitochondrial evolution. Science, 283, 1476–1481. Scholar
  12. 12.
    Gyorgy, B., Modos, K., Pallinger, E., Paloczi, K., Pasztoi, M., Misjak, P., Deli, M. A., Sipos, A., Szalai, A., Voszka, I., Polgar, A., Toth, K., Csete, M., Nagy, G., Gay, S., Falus, A., Kittel, A., & Buzas, E. I. (2011). Detection and isolation of cell-derived microparticles are compromised by protein complexes resulting from shared biophysical parameters. Blood, 117, e39–e48. Scholar
  13. 13.
    Hessvik, N. P., & Llorente, A. (2017). Current knowledge on exosome biogenesis and release. Cellular and Molecular Life Sciences, 75(2), 193-208.
  14. 14.
    Hezel, M. E. V., Nieuwland, R., Bruggen, R. V., & Juffermans, N. P. (2017). The ability of extracellular vesicles to induce a pro-inflammatory host response. International. Journal of Molecular Sciences, 18, 1285.
  15. 15.
    Iraci, N., Leonardi, T., Gessler, F., Vega, B., & Pluchino, S. (2016). Focus on extracellular vesicles: physiological role and signalling properties of extracellular membrane vesicles. International Journal of Molecular Sciences, 17, 171. Scholar
  16. 16.
    Kaza, A. K., Wamala, I., Friehs, I., Kuebler, J. D., Rathod, R. H., Berra, I., Ericsson, M., Yao, R., Thedsanamoorthy, J. K., Zurakowski, D., Levitsky, S., Del Nido, P. J., Cowan, D. B., & McCully, J. D. (2017). Myocardial rescue with autologous mitochondrial transplantation in a porcine model of ischemia/reperfusion. The Journal of Thoracic and Cardiovascular Surgery, 153, 934–943. Scholar
  17. 17.
    Kondratov, K., Kurapeev, D., Popov, M., Sidorova, M., Minasian, S., Galagudza, M., Kostareva, A., & Fedorov, A. (2016). Heparinase treatment of heparin-contaminated plasma from coronary artery bypass grafting patients enables reliable quantification of microRNAs. Biomolecular Detection and Quantification, 8, 9–14. Scholar
  18. 18.
    Kondratov, K. A., Petrova, T. A., Mikhailovskii, V. Y., Ivanova, A. N., Kostareva, A. A., & Fedorov, A. V. (2017). A study of extracellular vesicles isolated from blood plasma conducted by low-voltage scanning electron microscopy. Cell Tissue Biology, 11, 181–190. Scholar
  19. 19.
    McIlroy, D. J., Jarnicki, A. G., Au, G. G., Lott, N., Smith, D. W., Hansbro, P. M., & Balogh, Z. J. (2014). Mitochondrial DNA neutrophil extracellular traps are formed after trauma and subsequent surgery. Journal of Critical Care, 29(1133), e1131–e1135. Scholar
  20. 20.
    Miller, F. J., Rosenfeldt, F. L., Zhang, C., Linnane, A. W., & Nagley, P. (2003). Precise determination of mitochondrial DNA copy number in human skeletal and cardiac muscle by a PCR-based assay: lack of change of copy number with age. Nucleic Acids Research, 31, e61.CrossRefGoogle Scholar
  21. 21.
    Ong, S. B., Samangouei, P., Kalkhoran, S. B., & Hausenloy, D. J. (2015). The mitochondrial permeability transition pore and its role in myocardial ischemia reperfusion injury. Journal of Molecular and Cellular Cardiology, 78, 23–34. Scholar
  22. 22.
    Paunel-Gorgulu, A., Wacker, M., El Aita, M., Hassan, S., Schlachtenberger, G., Deppe, A., Choi, Y. H., Kuhn, E., Mehler, T. O., & Wahlers, T. (2017). cfDNA correlates with endothelial damage after cardiac surgery with prolonged cardiopulmonary bypass and amplifies NETosis in an intracellular TLR9-independent manner. Scientific Reports, 7, 17421. Scholar
  23. 23.
    Qin, C., Gu, J., Hu, J., Qian, H., Fei, X., Li, Y., Liu, R., & Meng, W. (2016). Platelets activation is associated with elevated plasma mitochondrial DNA during cardiopulmonary bypass. Journal of Cardiothoracic Surgery, 11, 90. Scholar
  24. 24.
    Raposo, G., & Stoorvogel, W. (2013). Extracellular vesicles: exosomes, microvesicles, and friends. The Journal of Cell Biology, 200, 373–383. Scholar
  25. 25.
    Rykova, E. Y., Morozkin, E. S., Ponomaryova, A. A., Loseva, E. M., Zaporozhchenko, I. A., Cherdyntseva, N. V., Vlassov, V. V., & Laktionov, P. P. (2012). Cell-free and cell-bound circulating nucleic acid complexes: mechanisms of generation, concentration and content. Expert Opinion on Biological Therapy, 12(Suppl 1), S141–S153. Scholar
  26. 26.
    Sandler, N., Kaczmarek, E., Itagaki, K., Zheng, Y., Otterbein, L., Khabbaz, K., Liu, D., Senthilnathan, V., Gruen, R. L., & Hauser, C. J. (2017). Mitochondrial DAMPs are released during cardiopulmonary bypass surgery and are associated with postoperative atrial fibrillation. Heart, Lung & Circulation, 27, 122–129. Scholar
  27. 27.
    Sluijter, J. P. G., Davidson, S. M., Boulanger, C. M., Buzas, E. I., de Kleijn, D. P. V., Engel, F. B., Giricz, Z., Hausenloy, D. J., Kishore, R., Lecour, S., Leor, J., Madonna, R., Perrino, C., Prunier, F., Sahoo, S., Schiffelers, R. M., Schulz, R., Van Laake, L. W., Ytrehus, K., & Ferdinandy, P. (2018). Extracellular vesicles in diagnostics and therapy of the ischaemic heart: Position Paper from the Working Group on Cellular Biology of the Heart of the European Society of Cardiology. Cardiovascular Research, 114, 19–34. Scholar
  28. 28.
    Svennerholm, K., Rodsand, P., Hellman, U., Waldenstrom, A., Lundholm, M., Ahren, D., Biber, B., Ronquist, G., & Haney, M. (2016). DNA content in extracellular vesicles isolated from porcine coronary venous blood directly after myocardial ischemic preconditioning. PLoS One, 11, e0159105. Scholar
  29. 29.
    van der Vaart, M., & Pretorius, P. J. (2008). Circulating DNA. Its origin and fluctuation. Annals of the New York Academy of Sciences, 1137, 18–26. Scholar
  30. 30.
    Vyas, N., Walvekar, A., Tate, D., Lakshmanan, V., Bansal, D., Lo Cicero, A., Raposo, G., Palakodeti, D., & Dhawan, J. (2014). Vertebrate hedgehog is secreted on two types of extracellular vesicles with different signaling properties. Scientific Reports, 4, 7357. Scholar
  31. 31.
    Waldenstrom, A., Genneback, N., Hellman, U., & Ronquist, G. (2012). Cardiomyocyte microvesicles contain DNA/RNA and convey biological messages to target cells. PLoS One, 7, e34653. Scholar
  32. 32.
    Witwer, K. W., Buzas, E. I., Bemis, L. T., Bora, A., Lasser, C., Lotvall, J., Nolte-‘t Hoen, E. N., Piper, M. G., Sivaraman, S., Skog, J., Thery, C., Wauben, M. H., & Hochberg, F. (2013). Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. Journal of Extracellular Vesicles, 2, 20360.
  33. 33.
    Zhang, Q., Itagaki, K., & Hauser, C. J. (2010). Mitochondrial DNA is released by shock and activates neutrophils via p38 map kinase. Shock, 34, 55–59. Scholar
  34. 34.
    Zhang, Q., Raoof, M., Chen, Y., Sumi, Y., Sursal, T., Junger, W., Brohi, K., Itagaki, K., & Hauser, C. J. (2010). Circulating mitochondrial DAMPs cause inflammatory responses to injury. Nature, 464, 104–107. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Anton Baysa
    • 1
    • 2
  • Anton Fedorov
    • 3
    • 4
  • Kirill Kondratov
    • 3
  • Arno Ruusalepp
    • 5
  • Sarkis Minasian
    • 3
  • Michael Galagudza
    • 3
  • Maxim Popov
    • 3
  • Dmitry Kurapeev
    • 3
  • Alexey Yakovlev
    • 3
  • Guro Valen
    • 1
    • 2
  • Anna Kostareva
    • 3
  • Jarle Vaage
    • 6
  • Kåre-Olav Stensløkken
    • 1
    • 2
    Email author
  1. 1.Division of Physiology, Department of Molecular Medicine, Institute of Basic Medical ScienceUniversity of OsloOsloNorway
  2. 2.Center for Heart Failure ResearchUniversity of OsloOsloNorway
  3. 3.Almazov National Medical Research CentreSaint PetersburgRussia
  4. 4.Department of Cytology and HistologySaint Petersburg State UniversitySaint PetersburgRussia
  5. 5.Department of Cardiac SurgeryTartu University HospitalTartuEstonia
  6. 6.Institute of Clinical Medicine, Oslo University HospitalUniversity of OsloOsloNorway

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