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Dynamics of leucocyte DNA thioguanine nucleotide levels during maintenance therapy of childhood acute lymphoblastic leukemia

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Abstract

Purpose

Methotrexate (MTX)/6-Mercaptopurine (6MP)-based maintenance therapy is crucial to cure childhood acute lymphoblastic leukemia (ALL). Cytotoxicity is mediated by incorporation of thioguanine nucleotides (TGN) into DNA (DNA-TG) with higher levels in leucocytes being associated with reduced relapse risk. To further understand the dynamics of DNA-TG formation, we measured DNA-TG levels in leucocyte subsets during maintenance therapy and in the months following its discontinuation.

Methods

DNA-TG levels were measured in leucocytes (DNA-TGTotal), polymorph nucleated granulocytes (neutrophils, eosinophils, basophils [DNA-TGPMN]) and mononucleated cells (lymphocytes, monocytes [DNA-TGMNC]) in 1013 samples from 52 patients on ALL maintenance therapy (951 samples during therapy and 62 samples after therapy discontinuation, respectively).

Results

Median DNA-TGTotal, DNA-TGPMN and DNA-TGMNC during maintenance therapy were 539, 563 and 384 fmol/µg DNA, respectively. DNA-TGPMN displayed more pronounced fluctuation than DNA-TGMNC (range 0–3084 [interquartile range IQR 271–881] versus 30–1411 [IQR 270–509] fmol/µg DNA). DNA-TGTotal was more strongly correlated with DNA-TGPMN (rS = 0.95, p < 0.0001) than DNA-TGMNC (rS = 0.73, p < 0.0001). DNA-TGPMN correlated less with DNA-TGMNC (rS = 0.64, p < 0.0001) and to a much lesser extent with absolute neutrophil count (rS = 0.35, p < 0.0001). Following discontinuation of therapy, DNA-TGPMN was rapidly eliminated, and not measurable beyond day 22 after discontinuation, whereas DNA-TGMNC was slowly eliminated, and five patients demonstrated a measurable DNA-TGMNC more than 365 days after therapy discontinuation.

Conclusion

Fluctuations in DNA-TGTotal are predominantly caused by corresponding fluctuations in DNA-TGPMN, thus DNA-TGTotal measures recent TGN incorporation in these short-lived cells. Measurement of DNA-TGTotal at 2–4 weeks intervals provides a reliable profile of DNA-TG levels.

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Acknowledgements

We thank the dedicated staff at the laboratory of Pediatric Oncology, Bonkolab, Copenhagen for their valuable work. This work is part of Childhood Oncology Network Targeting Research, Organisation & Life expectancy (CONTROL) and supported by Danish Cancer Society (R-257-A14720) and the Danish Childhood Cancer Foundation (2019-5934).

Funding

Danish Cancer Society, Danish Childhood Cancer Foundation, Novo Nordisk Foundation, Swedish Childhood Cancer Foundation, Nordic Cancer Union, The Capital Region of Denmark, and Rigshospitalet, University of Copenhagen.

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Authors and Affiliations

  1. Department of Pediatrics and Adolescent Medicine, Rigshospitalet, University of Copenhagen, Copenhagen, Denmark

    Rikke Hebo Larsen, Lisa Lyngsie Hjalgrim, Matilda Degn, Jacob Nersting, Bodil Als-Nielsen, Kathrine Grell & Kjeld Schmiegelow

  2. Section of Biostatistics, Department of Public Health, University of Copenhagen, Copenhagen, Denmark

    Kathrine Grell

  3. Institute of Clinical Medicine, Faculty of Medicine, University of Copenhagen, Copenhagen, Denmark

    Kjeld Schmiegelow

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  1. Rikke Hebo Larsen
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Contributions

RHL coordinated the study, compiled data, and drafted the manuscript. JN and MD performed DNA-TG analyses. KG supervised the study and performed the statistical analysis. LH and BAN supervised the study. KS initiated and supervised the study and had responsibility for the final submission for publication. All authors approved the final manuscript.

Corresponding author

Correspondence to Kjeld Schmiegelow.

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Conflict of interest

Kjeld Schmiegelow has received speaker and/or advisory board honoraria from Jazz Pharmaceuticals and Servier; speaker fee from Amgen and Medscape; Educational grant from Servier.

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The study was approved by the Ethical Committee of the Capital Region of Denmark (H-2.2010–002).

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Written informed consent was obtained from all participants or legal guardians.

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Larsen, R.H., Hjalgrim, L.L., Degn, M. et al. Dynamics of leucocyte DNA thioguanine nucleotide levels during maintenance therapy of childhood acute lymphoblastic leukemia. Cancer Chemother Pharmacol 88, 53–60 (2021). https://doi.org/10.1007/s00280-020-04219-5

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