Ascorbate Induces Ten-Eleven Translocation (Tet) Methylcytosine Dioxygenase-mediated Generation of 5-Hydroxymethylcytosine

Minor, Emily A.; Court, Brenda L.; Young, Juan I.; Wang, Gaofeng

doi:10.1074/jbc.c113.464800

Cited by 354 publications

(336 citation statements)

References 22 publications

(31 reference statements)

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“…Ascorbate has been shown to drive DNA demethylation in cultured cells in a TET-dependent manner (19,21,22,25); however, this effect was not measured in a genome-wide or fully quantitative fashion. We determined cytosine modification levels in nESCs using mass spectrometry and found that 50 μg/mL ascorbate supplementation over 72 h decreased 5mC by almost one half (1.7-fold reduction) and increased 5hmC by 2.8-fold (Fig.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Retinol and ascorbate drive erasure of epigenetic memory and enhance reprogramming to naïve pluripotency by complementary mechanisms

Hore

Meyenn

Ravichandran

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

148

139

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Epigenetic memory, in particular DNA methylation, is established during development in differentiating cells and must be erased to create naïve (induced) pluripotent stem cells. The ten-eleven translocation (TET) enzymes can catalyze the oxidation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and further oxidized derivatives, thereby actively removing this memory. Nevertheless, the mechanism by which the TET enzymes are regulated, and the extent to which they can be manipulated, are poorly understood. Here we report that retinoic acid (RA) or retinol (vitamin A) and ascorbate (vitamin C) act as modulators of TET levels and activity. RA or retinol enhances 5hmC production in naïve embryonic stem cells by activation of TET2 and TET3 transcription, whereas ascorbate potentiates TET activity and 5hmC production through enhanced Fe 2+ recycling, and not as a cofactor as reported previously. We find that both ascorbate and RA or retinol promote the derivation of induced pluripotent stem cells synergistically and enhance the erasure of epigenetic memory. This mechanistic insight has significance for the development of cell treatments for regenenerative medicine, and enhances our understanding of how intrinsic and extrinsic signals shape the epigenome.

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Section: Resultsmentioning

confidence: 99%

“…Nevertheless, the molecular signals that control TET activity in nESCs, and how they can be manipulated during reprogramming, are poorly characterized. For example, although ascorbate (vitamin C) is known to enhance 5hmC production in a TET-dependent manner (19)(20)(21)(22)(23), the mechanism by which this occurs is unclear (Fig. 1B).…”

mentioning

confidence: 99%

Retinol and ascorbate drive erasure of epigenetic memory and enhance reprogramming to naïve pluripotency by complementary mechanisms

Hore

Meyenn

Ravichandran

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

148

139

View full text Add to dashboard Cite

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“…Under this scenario, ascorbate would be better thought of as a salvager of TET catalysis following 'uncoupled' oxidation in the absence of substrate (which results in the production of Fe 3+ ), rather than a cofactor intrinsically required for its activity. Nevertheless, initial biochemical studies did not support this contention; reducing agents other than ascorbate were apparently insufficient to enhance TET activity in vitro [45,50,51,53]. Subsequent cell culture experiments using quinones as reducing agents [54,55], and further in vitro experiments at physiological pH [37] have now challenged this conclusion.…”

Section: Vitamin a Enhances Tet Expression Vitamin C Potentiates Tetmentioning

confidence: 93%

“…Ascorbate has been shown on numerous occasions to increase 5hmC and decrease 5mC in ESCs, however, in contrast to vitamin A, this occurs without any change in TET expression [37,45,[49][50][51]. Ascorbate is a well-characterized reducing agent which acts on Fe 3+ ions, and given that Fe 2+ is essential for TET activity, it was implicitly assumed that the antioxidative capacity of ascorbate is what accounts for increased 5hmC deposition in its presence [52].…”

Section: Vitamin a Enhances Tet Expression Vitamin C Potentiates Tetmentioning

confidence: 99%

Modulating Epigenetic Memory through Vitamins and TET: Implications for Regenerative Medicine and Cancer Treatment

Hore

2017

Epigenomics

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Vitamins A and C represent unrelated sets of small molecules that are essential to the human diet and have recently been shown to intensify erasure of epigenetic memory in naive embryonic stem cells. These effects are driven by complementary enhancement of the ten-eleven translocation (TET) demethylases -vitamin A stimulates TET expression, whereas vitamin C potentiates TET catalytic activity. Vitamin A and C cosupplementation synergistically enhances reprogramming of differentiated cells to the naive state, but overuse may exaggerate instability of imprinted genes. As such, optimizing their use in culture media will be important for regenerative medicine and mammalian transgenics. In addition, mechanistic perception of how these vitamins interact with the epigenome may be relevant for understanding cancer and improving patient treatment. Figure 1A), and both are associated with a long recorded history. Nightblindness, an early symptom of vitamin A deficiency, and its cure through liver consumption, was known to Egyptians in the prehistoric period [1]. Equally, the debilitating effects of scurvy (vitamin C deficiency) among sailors, and its treatment with oranges, was registered as far back as 1498 [2]. Vitamins A and C also have a lengthy and rich history in scientific literature; in 1753 James Lind described the first controlled clinical trial where various antiscorbutic treatments were tested on British seamen and in 1929, Frederick Gowland Hopkins was co-awarded the Nobel Prize in Medicine for the 'discovery of vitamins' following milk supplementation experiments in vitamin A deficient mice [3]. Eventual isolation of the respective compounds underlying both vitamins led to Nobel Prizes in Chemistry and Medicine in 1937 [4]. Since this time, a considerable amount of scientific progress (and at times conjecture) has been associated with vitamins A and C, and their respective biological effects touch disciplines as divergent as development and dermatology.One of the most recently discovered fields that vitamins A and C impact upon is epigenetics [5][6][7]. New research has shown that both vitamins drive active removal of DNA methylation in embryonic stem cells (ESCs) by enhancing the same family of ten-eleven translocation (TET) enzymes, and in doing so, help erase DNA methylation and the epigenetic memory encoded by it to improve the reprogramming of differentiated cells to an embryonic-like state. Here I review the effects of vitamins A and C on DNA methylation and epigenetic memory in stem cells, and outline their significance for the fields For reprint orders, please contact: reprints@futuremedicine.com

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“…1B). These results prompted us to search for unknown factors leading to the CNS2 demethylation of pTregs, and we tested vitamin C on the basis of reports that had showed a relationship between vitamin C and DNA demethylation (41)(42)(43)(44). Because 40260 mM (7.04210.56 mg/ml) vitamin C is detected in serum and RBCs (45,46), we cultured naive CD4 + T cells under iTreg conditions (anti-CD3/CD28, rIL2 and rTGF-b) with various doses of vitamin C (1, 10, and 100 mg/ml).…”

Section: Vitamin C Induced Cns2 Demethylation In Itregsmentioning

confidence: 99%

Vitamin C Facilitates Demethylation of the Foxp3 Enhancer in a Tet-Dependent Manner

Nair

Song

2016

The Journal of Immunology

147

100

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Demethylation of CpG motifs in the Foxp3 intronic element, conserved noncoding sequence 2 (CNS2), is indispensable for the stable expression of Foxp3 in regulatory T cells (Tregs). In this study, we found that vitamin C induces CNS2 demethylation in Tregs in a ten-eleven-translocation 2 (Tet2)-dependent manner. The CpG motifs of CNS2 in Tregs generated in vitro by TGF-β (iTregs), which were methylated originally, became demethylated after vitamin C treatment. The conversion of 5-methylcytosin into 5-hydroxymethylcytosin was more efficient, and the methyl group from the CpG motifs of Foxp3 CNS2 was erased rapidly in iTregs treated with vitamin C. The effect of vitamin C disappeared in Tet2−/− iTregs. Furthermore, CNS2 in peripheral Tregs in vivo, which were demethylated originally, became methylated after treatment with a sodium-dependent vitamin C transporter inhibitor, sulfinpyrazone. Finally, CNS2 demethylation in thymic Tregs was also impaired in Tet2−/− mice, but not in wild type mice, when they were treated with sulfinpyrazone. Collectively, vitamin C was required for the CNS2 demethylation mediated by Tet proteins, which was essential for Foxp3 expression. Our findings indicate that environmental factors, such as nutrients, could bring about changes in immune homeostasis through epigenetic mechanisms.

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Ascorbate Induces Ten-Eleven Translocation (Tet) Methylcytosine Dioxygenase-mediated Generation of 5-Hydroxymethylcytosine

Cited by 354 publications

References 22 publications

Retinol and ascorbate drive erasure of epigenetic memory and enhance reprogramming to naïve pluripotency by complementary mechanisms

Retinol and ascorbate drive erasure of epigenetic memory and enhance reprogramming to naïve pluripotency by complementary mechanisms

Modulating Epigenetic Memory through Vitamins and TET: Implications for Regenerative Medicine and Cancer Treatment

Vitamin C Facilitates Demethylation of the Foxp3 Enhancer in a Tet-Dependent Manner

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