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Oxidative Medicine and Cellular Longevity
Volume 2019, Article ID 8156592, 11 pages
https://doi.org/10.1155/2019/8156592
Research Article

Decreased Nuclear Ascorbate Accumulation Accompanied with Altered Genomic Methylation Pattern in Fibroblasts from Arterial Tortuosity Syndrome Patients

1Department of Medical Chemistry, Molecular Biology, and Pathobiochemistry, Semmelweis University, Budapest 1094, Hungary
2Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest 1117, Hungary
3Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Budapest 1117, Hungary
4Center for Medical Genetics, Ghent University, Ghent B-9000, Belgium
5Division of Biology and Genetics, Department of Molecular and Translational Medicine, University of Brescia, Brescia 25123, Italy
6Pathobiochemistry Research Group of Hungarian Academy of Sciences & Semmelweis University, Budapest 1094, Hungary
7Institute of Clinical Experimental Research, Semmelweis University, Budapest 1094, Hungary

Correspondence should be addressed to Éva Margittai; uh.vinu-siewlemmes.dem@ave.iattigram

Received 26 July 2018; Accepted 28 November 2018; Published 13 January 2019

Academic Editor: Olga Pechanova

Copyright © 2019 Csilla E. Németh et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Ascorbate requiring Fe2+/2-oxoglutarate-dependent dioxygenases located in the nucleoplasm have been shown to participate in epigenetic regulation of gene expression via histone and DNA demethylation. Transport of dehydroascorbic acid is impaired in the endomembranes of fibroblasts from arterial tortuosity syndrome (ATS) patients, due to the mutation in the gene coding for glucose transporter GLUT10. We hypothesized that altered nuclear ascorbate concentration might be present in ATS fibroblasts, affecting dioxygenase activity and DNA demethylation. Therefore, our aim was to characterize the subcellular distribution of vitamin C, the global and site-specific changes in 5-methylcytosine and 5-hydroxymethylcytosine levels, and the effect of ascorbate supplementation in control and ATS fibroblast cultures. Diminished nuclear accumulation of ascorbate was found in ATS fibroblasts upon ascorbate or dehydroascorbic acid addition. Analyzing DNA samples of cultured fibroblasts from controls and ATS patients, a lower global 5-hydroxymethylcytosine level was found in ATS fibroblasts, which could not be significantly modified by ascorbate addition. Investigation of the (hydroxy)methylation status of specific regions in six candidate genes related to ascorbate metabolism and function showed that ascorbate addition could stimulate hydroxymethylation and active DNA demethylation at the PPAR-γ gene region in control fibroblasts only. The altered DNA hydroxymethylation patterns in patient cells both at the global level and at specific gene regions accompanied with decreased nuclear accumulation of ascorbate suggests the epigenetic role of vitamin C in the pathomechanism of ATS. The present findings represent the first example for the role of vitamin C transport in epigenetic regulation suggesting that ATS is a compartmentalization disease.