title: Urinary Measurement of Epigenetic DNA Modifications: A Non-Invasive Assessment of the Whole-Body Epigenetic Status in Healthy Subjects and Colorectal Cancer Patients.
abstract: Active mechanism of DNA demethylation can be responsible for the activation of previously silenced genes. Products of 5-methylcytosine oxidation are released into the bloodstream and eventually excreted with urine. Therefore, whole-body epigenetic status can be assessed non-invasively on the basis of the urinary excretion of a broad spectrum of epigenetic modifications: 5-hydroxymethylcytosine (5-hmCyt), 5-formylcytosine (5-fCyt), 5-carboxycytosine (5-caCyt), and 5-hydroxymethyluracil (5-hmUra). We have developed a specific and sensitive, isotope-dilution, automated, online, two-dimensional ultra-performance liquid chromatography system with tandem mass spectrometry (2D UPLC-MS/MS) to measure 5-hmCyt, 5-fCyt, 5-caCyt, and their deoxynucleosides in the same urine sample. Human urine contains all of the modifications except from 5-formyl-2'-deoxycytidine (5-fdC) and 5-carboxy-2'-deoxycytidine (5-cadC). A highly significant difference in the urinary excretion of 5-(hydroxymethyl)-2'-deoxycytidine (5-hmdC) was found between healthy subjects and colorectal cancer patients (3.5 vs. 7.8 nmol mmol-1creatinine, respectively), as well as strong correlations between the majority of analyzed compounds.
authors.affiliation: Department of Clinical Biochemistry Faculty of Pharmacy Nicolaus Copernicus University in Toruń Collegium Medicum in Bydgoszcz Karlowicza 24 85-092 Bydgoszcz Poland.
authors.forename: Rafal
authors.identifier: 0000-0003-2067-9981
authors.initials: R
authors.lastname: Rozalski
authors.affiliation: Department of Clinical Biochemistry Faculty of Pharmacy Nicolaus Copernicus University in Toruń Collegium Medicum in Bydgoszcz Karlowicza 24 85-092 Bydgoszcz Poland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Gackowski
authors.affiliation: Department of Clinical Biochemistry Faculty of Pharmacy Nicolaus Copernicus University in Toruń Collegium Medicum in Bydgoszcz Karlowicza 24 85-092 Bydgoszcz Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Siomek-Gorecka
authors.affiliation: Department of Surgery Faculty of Medicine Nicolaus Copernicus University in Toruń Collegium Medicum in Bydgoszcz Ujejskiego 75 85-168 Bydgoszcz Poland.
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Banaszkiewicz
authors.affiliation: Department of Clinical Biochemistry Faculty of Pharmacy Nicolaus Copernicus University in Toruń Collegium Medicum in Bydgoszcz Karlowicza 24 85-092 Bydgoszcz Poland.
authors.forename: Ryszard
authors.initials: R
authors.lastname: Olinski
keywords: 2D UPLC–MS/MS; DNA damage; DNA methylation; epigenetics; urine