Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes

Ori Fridlich; Ayelet Peretz; Ilana Fox-Fisher; Sheina Pyanzin; Ziv Dadon; Eilon Shcolnik; Ronen Sadeh; Gavriel Fialkoff; Israa Sharkia; Joshua Moss; Ludovica Arpinati; Shachar Nice; Christopher D. Nogiec; Samuel Terkper Ahuno; Rui Li; Eddie Taborda; Sonia Dunkelbarger; Zvi G. Fridlender; Paz Polak; Tommy Kaplan; Nir Friedman; Benjamin Glaser; Ruth Shemer; Naama Constantini; Yuval Dor

doi:10.1016/j.xcrm.2023.101074

Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes

Ori Fridlich, Ayelet Peretz, Ilana Fox-Fisher, Sheina Pyanzin, Ziv Dadon, Eilon Shcolnik, Ronen Sadeh, Gavriel Fialkoff, Israa Sharkia, Joshua Moss, Ludovica Arpinati, Shachar Nice, Christopher D. Nogiec, Samuel Terkper Ahuno, Rui Li, Eddie Taborda, Sonia Dunkelbarger, Zvi G. Fridlender, Paz Polak, Tommy KaplanNir Friedman, Benjamin Glaser, Ruth Shemer, Naama Constantini, Yuval Dor^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Strenuous physical exercise causes a massive elevation in the concentration of circulating cell-free DNA (cfDNA), which correlates with effort intensity and duration. The cellular sources and physiological drivers of this phenomenon are unknown. Using methylation patterns of cfDNA and associated histones, we show that cfDNA in exercise originates mostly in extramedullary polymorphonuclear neutrophils. Strikingly, cardiomyocyte cfDNA concentration increases after a marathon, consistent with elevated troponin levels and indicating low-level, delayed cardiac cell death. Physical impact, low oxygen levels, and elevated core body temperature contribute to neutrophil cfDNA release, while muscle contraction, increased heart rate, β-adrenergic signaling, or steroid treatment fail to cause elevation of cfDNA. Physical training reduces neutrophil cfDNA release after a standard exercise, revealing an inverse relationship between exercise-induced cfDNA release and training level. We speculate that the release of cfDNA from neutrophils in exercise relates to the activation of neutrophils in the context of exercise-induced muscle damage.

Original language	American English
Article number	101074
Journal	Cell Reports Medicine
Volume	4
Issue number	6
DOIs	https://doi.org/10.1016/j.xcrm.2023.101074
State	Published - 20 Jun 2023

Bibliographical note

Funding Information:
This work was supported by grants from the Beutler Research Program of Excellence in Genomic Medicine, The Israel Science Foundation , the Waldholtz/Pakula family, the Helmsley Charitable Trust and Grail (to Y.D.), and the European Research Council (cfChIP, to N.F.). Y.D. holds the Walter and Greta Stiel Chair and Research Grant in Heart Studies. N.C. received financial support for this project from Shaare Zedek Medical Center .

Funding Information:
We thank Keren Constantini for stimulating discussions and data collection, Dana and Ibrahim Deeb for help in blood collection, Elior Shaked and Netta Kasher for graphic visualization, and Miriam Ravins and Emanuel Hanski for advice and generous support in exercise plasma experiments. We thank Idit Shiff and Abed Nasereddin from the Core Research Facility (CRF) at the Hebrew University Faculty of Medicine. We are grateful to the volunteers who donated blood. This work was supported by grants from the Beutler Research Program of Excellence in Genomic Medicine, The Israel Science Foundation, the Waldholtz/Pakula family, the Helmsley Charitable Trust and Grail (to Y.D.), and the European Research Council (cfChIP, to N.F.). Y.D. holds the Walter and Greta Stiel Chair and Research Grant in Heart Studies. N.C. received financial support for this project from Shaare Zedek Medical Center. Y.D. N.C. R.S. O.F. B.G. and Z.D. designed experiments. T.K. and J.M. performed computational analysis of cfDNA methylomes. I.F.-F. established immune cell methylation markers. O.F. A.P. S.P. Z.D. E.S. and S.N. performed experiments. G.F. R. Sadeh, I.S. and N.F. performed and analyzed cfCHIP-seq experiments. L.A. and Z.G.F. contributed to NETs experiments. C.N. S.T.A. R.L. E.T. S.D. and P.P. designed and performed endurance versus resistance experiments. O.F. N.C. and Y.D. wrote the paper. R. Sadeh, G.F. I.S. and N.F. are founders and/or employees of Senseera Inc. J.M. B.G. R. Shemer, and Y.D. have filed patents on cfDNA methylation analysis.

Publisher Copyright:
© 2023 The Author(s)

Keywords

ChIP-seq
chromatin
circulating cell-free DNA
exercise biology
fitness
inflammation
methylation
neutrophil extracellular traps
neutrophils
polymorphonuclear cells

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10.1016/j.xcrm.2023.101074

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Fridlich, O., Peretz, A., Fox-Fisher, I., Pyanzin, S., Dadon, Z., Shcolnik, E., Sadeh, R., Fialkoff, G., Sharkia, I., Moss, J., Arpinati, L., Nice, S., Nogiec, C. D., Ahuno, S. T., Li, R., Taborda, E., Dunkelbarger, S., Fridlender, Z. G., Polak, P., ... Dor, Y. (2023). Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes. Cell Reports Medicine, 4(6), Article 101074. https://doi.org/10.1016/j.xcrm.2023.101074

@article{ed26aed09d47458798579e67da1e8d15,

title = "Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes",

abstract = "Strenuous physical exercise causes a massive elevation in the concentration of circulating cell-free DNA (cfDNA), which correlates with effort intensity and duration. The cellular sources and physiological drivers of this phenomenon are unknown. Using methylation patterns of cfDNA and associated histones, we show that cfDNA in exercise originates mostly in extramedullary polymorphonuclear neutrophils. Strikingly, cardiomyocyte cfDNA concentration increases after a marathon, consistent with elevated troponin levels and indicating low-level, delayed cardiac cell death. Physical impact, low oxygen levels, and elevated core body temperature contribute to neutrophil cfDNA release, while muscle contraction, increased heart rate, β-adrenergic signaling, or steroid treatment fail to cause elevation of cfDNA. Physical training reduces neutrophil cfDNA release after a standard exercise, revealing an inverse relationship between exercise-induced cfDNA release and training level. We speculate that the release of cfDNA from neutrophils in exercise relates to the activation of neutrophils in the context of exercise-induced muscle damage.",

keywords = "ChIP-seq, chromatin, circulating cell-free DNA, exercise biology, fitness, inflammation, methylation, neutrophil extracellular traps, neutrophils, polymorphonuclear cells",

author = "Ori Fridlich and Ayelet Peretz and Ilana Fox-Fisher and Sheina Pyanzin and Ziv Dadon and Eilon Shcolnik and Ronen Sadeh and Gavriel Fialkoff and Israa Sharkia and Joshua Moss and Ludovica Arpinati and Shachar Nice and Nogiec, {Christopher D.} and Ahuno, {Samuel Terkper} and Rui Li and Eddie Taborda and Sonia Dunkelbarger and Fridlender, {Zvi G.} and Paz Polak and Tommy Kaplan and Nir Friedman and Benjamin Glaser and Ruth Shemer and Naama Constantini and Yuval Dor",

note = "Funding Information: This work was supported by grants from the Beutler Research Program of Excellence in Genomic Medicine, The Israel Science Foundation , the Waldholtz/Pakula family, the Helmsley Charitable Trust and Grail (to Y.D.), and the European Research Council (cfChIP, to N.F.). Y.D. holds the Walter and Greta Stiel Chair and Research Grant in Heart Studies. N.C. received financial support for this project from Shaare Zedek Medical Center . Funding Information: We thank Keren Constantini for stimulating discussions and data collection, Dana and Ibrahim Deeb for help in blood collection, Elior Shaked and Netta Kasher for graphic visualization, and Miriam Ravins and Emanuel Hanski for advice and generous support in exercise plasma experiments. We thank Idit Shiff and Abed Nasereddin from the Core Research Facility (CRF) at the Hebrew University Faculty of Medicine. We are grateful to the volunteers who donated blood. This work was supported by grants from the Beutler Research Program of Excellence in Genomic Medicine, The Israel Science Foundation, the Waldholtz/Pakula family, the Helmsley Charitable Trust and Grail (to Y.D.), and the European Research Council (cfChIP, to N.F.). Y.D. holds the Walter and Greta Stiel Chair and Research Grant in Heart Studies. N.C. received financial support for this project from Shaare Zedek Medical Center. Y.D. N.C. R.S. O.F. B.G. and Z.D. designed experiments. T.K. and J.M. performed computational analysis of cfDNA methylomes. I.F.-F. established immune cell methylation markers. O.F. A.P. S.P. Z.D. E.S. and S.N. performed experiments. G.F. R. Sadeh, I.S. and N.F. performed and analyzed cfCHIP-seq experiments. L.A. and Z.G.F. contributed to NETs experiments. C.N. S.T.A. R.L. E.T. S.D. and P.P. designed and performed endurance versus resistance experiments. O.F. N.C. and Y.D. wrote the paper. R. Sadeh, G.F. I.S. and N.F. are founders and/or employees of Senseera Inc. J.M. B.G. R. Shemer, and Y.D. have filed patents on cfDNA methylation analysis. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = jun,

day = "20",

doi = "10.1016/j.xcrm.2023.101074",

language = "American English",

volume = "4",

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Fridlich, O, Peretz, A, Fox-Fisher, I, Pyanzin, S, Dadon, Z, Shcolnik, E, Sadeh, R, Fialkoff, G, Sharkia, I, Moss, J, Arpinati, L, Nice, S, Nogiec, CD, Ahuno, ST, Li, R, Taborda, E, Dunkelbarger, S, Fridlender, ZG, Polak, P, Kaplan, T, Friedman, N, Glaser, B, Shemer, R, Constantini, N & Dor, Y 2023, 'Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes', Cell Reports Medicine, vol. 4, no. 6, 101074. https://doi.org/10.1016/j.xcrm.2023.101074

TY - JOUR

T1 - Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes

AU - Fridlich, Ori

AU - Peretz, Ayelet

AU - Fox-Fisher, Ilana

AU - Pyanzin, Sheina

AU - Dadon, Ziv

AU - Shcolnik, Eilon

AU - Sadeh, Ronen

AU - Fialkoff, Gavriel

AU - Sharkia, Israa

AU - Moss, Joshua

AU - Arpinati, Ludovica

AU - Nice, Shachar

AU - Nogiec, Christopher D.

AU - Ahuno, Samuel Terkper

AU - Li, Rui

AU - Taborda, Eddie

AU - Dunkelbarger, Sonia

AU - Fridlender, Zvi G.

AU - Polak, Paz

AU - Kaplan, Tommy

AU - Friedman, Nir

AU - Glaser, Benjamin

AU - Shemer, Ruth

AU - Constantini, Naama

AU - Dor, Yuval

N1 - Funding Information: This work was supported by grants from the Beutler Research Program of Excellence in Genomic Medicine, The Israel Science Foundation , the Waldholtz/Pakula family, the Helmsley Charitable Trust and Grail (to Y.D.), and the European Research Council (cfChIP, to N.F.). Y.D. holds the Walter and Greta Stiel Chair and Research Grant in Heart Studies. N.C. received financial support for this project from Shaare Zedek Medical Center . Funding Information: We thank Keren Constantini for stimulating discussions and data collection, Dana and Ibrahim Deeb for help in blood collection, Elior Shaked and Netta Kasher for graphic visualization, and Miriam Ravins and Emanuel Hanski for advice and generous support in exercise plasma experiments. We thank Idit Shiff and Abed Nasereddin from the Core Research Facility (CRF) at the Hebrew University Faculty of Medicine. We are grateful to the volunteers who donated blood. This work was supported by grants from the Beutler Research Program of Excellence in Genomic Medicine, The Israel Science Foundation, the Waldholtz/Pakula family, the Helmsley Charitable Trust and Grail (to Y.D.), and the European Research Council (cfChIP, to N.F.). Y.D. holds the Walter and Greta Stiel Chair and Research Grant in Heart Studies. N.C. received financial support for this project from Shaare Zedek Medical Center. Y.D. N.C. R.S. O.F. B.G. and Z.D. designed experiments. T.K. and J.M. performed computational analysis of cfDNA methylomes. I.F.-F. established immune cell methylation markers. O.F. A.P. S.P. Z.D. E.S. and S.N. performed experiments. G.F. R. Sadeh, I.S. and N.F. performed and analyzed cfCHIP-seq experiments. L.A. and Z.G.F. contributed to NETs experiments. C.N. S.T.A. R.L. E.T. S.D. and P.P. designed and performed endurance versus resistance experiments. O.F. N.C. and Y.D. wrote the paper. R. Sadeh, G.F. I.S. and N.F. are founders and/or employees of Senseera Inc. J.M. B.G. R. Shemer, and Y.D. have filed patents on cfDNA methylation analysis. Publisher Copyright: © 2023 The Author(s)

PY - 2023/6/20

Y1 - 2023/6/20

N2 - Strenuous physical exercise causes a massive elevation in the concentration of circulating cell-free DNA (cfDNA), which correlates with effort intensity and duration. The cellular sources and physiological drivers of this phenomenon are unknown. Using methylation patterns of cfDNA and associated histones, we show that cfDNA in exercise originates mostly in extramedullary polymorphonuclear neutrophils. Strikingly, cardiomyocyte cfDNA concentration increases after a marathon, consistent with elevated troponin levels and indicating low-level, delayed cardiac cell death. Physical impact, low oxygen levels, and elevated core body temperature contribute to neutrophil cfDNA release, while muscle contraction, increased heart rate, β-adrenergic signaling, or steroid treatment fail to cause elevation of cfDNA. Physical training reduces neutrophil cfDNA release after a standard exercise, revealing an inverse relationship between exercise-induced cfDNA release and training level. We speculate that the release of cfDNA from neutrophils in exercise relates to the activation of neutrophils in the context of exercise-induced muscle damage.

AB - Strenuous physical exercise causes a massive elevation in the concentration of circulating cell-free DNA (cfDNA), which correlates with effort intensity and duration. The cellular sources and physiological drivers of this phenomenon are unknown. Using methylation patterns of cfDNA and associated histones, we show that cfDNA in exercise originates mostly in extramedullary polymorphonuclear neutrophils. Strikingly, cardiomyocyte cfDNA concentration increases after a marathon, consistent with elevated troponin levels and indicating low-level, delayed cardiac cell death. Physical impact, low oxygen levels, and elevated core body temperature contribute to neutrophil cfDNA release, while muscle contraction, increased heart rate, β-adrenergic signaling, or steroid treatment fail to cause elevation of cfDNA. Physical training reduces neutrophil cfDNA release after a standard exercise, revealing an inverse relationship between exercise-induced cfDNA release and training level. We speculate that the release of cfDNA from neutrophils in exercise relates to the activation of neutrophils in the context of exercise-induced muscle damage.

KW - ChIP-seq

KW - chromatin

KW - circulating cell-free DNA

KW - exercise biology

KW - fitness

KW - inflammation

KW - methylation

KW - neutrophil extracellular traps

KW - neutrophils

KW - polymorphonuclear cells

UR - http://www.scopus.com/inward/record.url?scp=85162165886&partnerID=8YFLogxK

U2 - 10.1016/j.xcrm.2023.101074

DO - 10.1016/j.xcrm.2023.101074

M3 - Article

C2 - 37290439

AN - SCOPUS:85162165886

SN - 2666-3791

VL - 4

JO - Cell Reports Medicine

JF - Cell Reports Medicine

IS - 6

M1 - 101074

ER -

Elevated cfDNA after exercise is derived primarily from mature polymorphonuclear neutrophils, with a minor contribution of cardiomyocytes

Abstract

Bibliographical note

Keywords

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