Dataset Information

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^-1 creatinine, respectively), as well as strong correlations between the majority of analyzed compounds.

SUBMITTER: Rozalski R

PROVIDER: S-EPMC5167278 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Urinary Measurement of Epigenetic DNA Modifications: A Non-Invasive Assessment of the Whole-Body Epigenetic Status in Healthy Subjects and Colorectal Cancer Patients.

Rozalski Rafal R Gackowski Daniel D Siomek-Gorecka Agnieszka A Banaszkiewicz Zbigniew Z Olinski Ryszard R

ChemistryOpen 20161115 6

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). ...[more]

PMID: 28032024

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Project description:BackgroundMaximal inspiratory pressure (MIP) is currently the most commonly used measure for respiratory muscle strength (RMS) estimation, however, requires significant effort. Falsely low values are therefore common, especially in fatigue-prone subjects, such as neuromuscular disorder patients. In contrast, sniff nasal inspiratory pressure (SNIP) requires a short, sharp sniff; this is a natural manoeuvre, decreasing required effort. Consequently, it has been suggested that use of SNIP could confirm the accuracy of MIP measurements. However, no recent guidelines regarding the optimal method of SNIP measurement exist, and varied approaches have been described.ObjectivesWe compared SNIP values from three conditions, namely with 30, 60 or 90 s time intervals between repeats, the right (SNIPR) and left (SNIPL) nostril, and the contralateral nostril occluded (SNIPO) or non-occluded (SNIPNO). Additionally, we determined the optimal number of repeats for accurate SNIP measurement.Method52 healthy subjects (23 males) were recruited for this study, of which a subset of 10 subjects (5 males) completed tests comparing the time interval between repeats. SNIP was measured from functional residual capacity via a probe in one nostril, while MIP was measured from residual volume.ResultsThere was no significant difference in SNIP depending on the interval between repeats (P = 0.98); subjects preferred the 30 s. SNIPO was significantly higher than SNIPNO (P < 0.00001) but SNIPL and SNIPR did not significantly differ (P = 0.60). There was an initial learning effect for the first SNIP test; SNIP did not decline during 80 repeats (P = 0.64).ConclusionsWe conclude that SNIPO is a more reliable RMS indicator than SNIPNO, as there is reduced risk of RMS underestimation. Allowing subjects to choose which nostril to use is appropriate, as this did not significantly affect SNIP, but may increase ease of performance. We suggest that twenty repeats is sufficient to overcome any learning effect and that fatigue is unlikely after this number of repeats. We believe these results are important in aiding the accurate collection of SNIP reference value data in the healthy population.

Project description:We undertook this study to understand how the transcription factor Sox2 contributes to the malignant phenotype of glioblastoma multiforme (GBM), the most aggressive primary brain tumor. We initially looked for unbalanced genomic rearrangements in the Sox2 locus in 42 GBM samples and found that Sox2 was amplified in 11.5% and overexpressed in all the samples. These results prompted us to further investigate the mechanisms involved in Sox2 overexpression in GBM. We analyzed the methylation status of the Sox2 promoter because high CpG density promoters are associated with key developmental genes. The Sox2 promoter presented a CpG island that was hypomethylated in all the patient samples when compared to normal cell lines. Treatment of Sox2-negative glioma cell lines with 5-azacitidine resulted in the re-expression of Sox2 and in a change in the methylation status of the Sox2 promoter. We further confirmed these results by analyzing data from GBM cases generated by The Cancer Genome Atlas project. We observed Sox2 overexpression (86%; N = 414), Sox2 gene amplification (8.5%; N = 492), and Sox 2 promoter hypomethylation (100%; N = 258), suggesting the relevance of this factor in the malignant phenotype of GBMs. To further explore the role of Sox2, we performed in vitro analysis with brain tumor stem cells (BTSCs) and established glioma cell lines. Downmodulation of Sox2 in BTSCs resulted in the loss of their self-renewal properties. Surprisingly, ectopic expression of Sox2 in established glioma cells was not sufficient to support self-renewal, suggesting that additional factors are required. Furthermore, we observed that ectopic Sox2 expression was sufficient to induce invasion and migration of glioma cells, and knockdown experiments demonstrated that Sox2 was essential for maintaining these properties. Altogether, our data underscore the importance of a pleiotropic role of Sox2 and suggest that it could be used as a therapeutic target in GBM.

Dataset Information

Urinary Measurement of Epigenetic DNA Modifications: A Non-Invasive Assessment of the Whole-Body Epigenetic Status in Healthy Subjects and Colorectal Cancer Patients.

Publications

Urinary Measurement of Epigenetic DNA Modifications: A Non-Invasive Assessment of the Whole-Body Epigenetic Status in Healthy Subjects and Colorectal Cancer Patients.

Similar Datasets

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