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. 2021 Nov 1;11(1):21345.
doi: 10.1038/s41598-021-00880-9.

The urinary excretion of epigenetically modified DNA as a marker of pediatric ALL status and chemotherapy response

Rafal Rozalski  1 , Daniel Gackowski  2 , Aleksandra Skalska-Bugala  2 , Marta Starczak  2 , Agnieszka Siomek-Gorecka  2 , Ewelina Zarakowska  2 , Martyna Modrzejewska  2 , Tomasz Dziaman  2 , Anna Szpila  2 , Kinga Linowiecka  2   3 , Jolanta Guz  2 , Justyna Szpotan  2   3 , Maciej Gawronski  2 , Anna Labejszo  2   4 , Lidia Gackowska  5 , Marek Foksinski  2 , Elwira Olinska  6 , Aleksandra Wasilow  2 , Andrzej Koltan  7 , Jan Styczynski  7 , Ryszard Olinski  8
Affiliations

The urinary excretion of epigenetically modified DNA as a marker of pediatric ALL status and chemotherapy response

Rafal Rozalski et al. Sci Rep. .

Abstract

The active DNA demethylation process may be linked to aberrant methylation and may be involved in leukemogenesis. We investigated the role of epigenetic DNA modifications in childhood acute lymphoblastic leukemia (ALL) diagnostics and therapy monitoring. We analyzed the levels of 5-methyl-2'-deoxycytidine (5-mdC) oxidation products in the cellular DNA and urine of children with ALL (at diagnosis and during chemotherapy, n = 55) using two-dimensional ultra-performance liquid chromatography with tandem mass spectrometry (2D UPLC-MS/MS). Moreover, the expression of Ten Eleven Translocation enzymes (TETs) at the mRNA and protein levels was determined. Additionally, the ascorbate level in the blood plasma was analyzed. Before treatment, the ALL patients had profoundly higher levels of the analyzed modified DNA in their urine than the controls. After chemotherapy, we observed a statistically significant decrease in active demethylation products in urine, with a final level similar to the level characteristic of healthy children. The level of 5-hmdC in the DNA of the leukocytes in blood of the patient group was significantly lower than that of the control group. Our data suggest that urinary excretion of epigenetic DNA modification may be a marker of pediatric ALL status and a reliable marker of chemotherapy response.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
The cytosine methylation and active demethylation pathway. (DNMT4: DNA methyltransferase, TETs: ten eleven translocation enzymes, AID: activation-induced cytosine deaminase, APOBEC: apolipoprotein B mRNA editing enzyme, TDG: thymine DNA glycosylase, SMUG1: single-strand-selective monofunctional uracil-DNA glycosylase 1, UNG: uracil-DNA glycosylase, MBD4: methyl-CpG-binding domain protein 4, BER: base excision repair). Reprinted from Biochimica et Biophysica Acta—Reviews on Cancer, 1869 (1), Olinski R, Gackowski D, Cooke M. Endogenously generated DNA nucleobase modifications source, and significance as possible biomarkers of malignant transformation risk, and role in anticancer therapy, 29–41 (2018), with permission from Elsevier.
Figure 2
Figure 2
Receiver operating characteristic (ROC) curves for epigenetically modified DNA markers in urine (∗p<0.05). AUC area under the curve.
Figure 3
Figure 3
Correlation between 5-hydroxymethylcytosine and % blasts at diagnosis.
Figure 4
Figure 4
Urinary levels of DNA damage markers and active demethylation products of 5-methylcytosine in healthy controls and ALL patients before therapy (A), 33 days after treatment (B) and six months after treatment (C). The results are presented as median values (∗p<0.05).
Figure 5
Figure 5
Ascorbic acid in plasma.
See this image and copyright information in PMC

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