title: Retinol and ascorbate drive erasure of epigenetic memory and enhance reprogramming to naïve pluripotency by complementary mechanisms.
abstract: 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 Fe2+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.
authors.affiliation: Epigenetics Programme, Babraham Institute, Cambridge CB22 3AT, United Kingdom; Department of Anatomy, University of Otago, Dunedin 9016, New Zealand; tim.hore@otago.ac.nz tomasz.jurkowski@ibc.uni-stuttgart.de wolf.reik@babraham.ac.uk.
authors.forename: Timothy Alexander
authors.identifier: 0000-0002-6735-225X
authors.initials: TA
authors.lastname: Hore
authors.affiliation: Epigenetics Programme, Babraham Institute, Cambridge CB22 3AT, United Kingdom.
authors.forename: Ferdinand
authors.initials: F
authors.lastname: von Meyenn
authors.affiliation: Institute of Biochemistry, University of Stuttgart, 70569 Stuttgart, Germany.
authors.forename: Mirunalini
authors.initials: M
authors.lastname: Ravichandran
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom; Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, United Kingdom.
authors.forename: Martin
authors.identifier: 0000-0003-3903-6492
authors.initials: M
authors.lastname: Bachman
authors.affiliation: Barts Cancer Institute, Queen Mary University of London, London EC1M 6BQ, United Kingdom.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Ficz
authors.affiliation: Mass Spectrometry Facility, Babraham Institute, Cambridge CB22 3AT, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Oxley
authors.affiliation: Epigenetics Programme, Babraham Institute, Cambridge CB22 3AT, United Kingdom.
authors.forename: Fátima
authors.initials: F
authors.lastname: Santos
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom; Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge CB2 0RE, United Kingdom.
authors.forename: Shankar
authors.initials: S
authors.lastname: Balasubramanian
authors.affiliation: Institute of Biochemistry, University of Stuttgart, 70569 Stuttgart, Germany; tim.hore@otago.ac.nz tomasz.jurkowski@ibc.uni-stuttgart.de wolf.reik@babraham.ac.uk.
authors.forename: Tomasz P
authors.initials: TP
authors.lastname: Jurkowski
authors.affiliation: Epigenetics Programme, Babraham Institute, Cambridge CB22 3AT, United Kingdom; Wellcome Trust Sanger Institute, Hinxton CB10 1SA, United Kingdom tim.hore@otago.ac.nz tomasz.jurkowski@ibc.uni-stuttgart.de wolf.reik@babraham.ac.uk.
authors.forename: Wolf
authors.initials: W
authors.lastname: Reik
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D019175:DNA Methylation / Q000187:drug effects / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D049308:Dioxygenases / Q000235:genetics*; D044127:Epigenesis, Genetic / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D044968:Regenerative Medicine; D014212:Tretinoin / Q000494:pharmacology; D014801:Vitamin A / Q000494:pharmacology
chemical_list: D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D014801:Vitamin A; D014212:Tretinoin; D044503:5-Methylcytosine; C578792:TET3 protein, human; D049308:Dioxygenases; C541081:TET2 protein, human; D001205:Ascorbic Acid
keywords: DNA methylation; TET; epigenetic memory; naive pluripotency; vitamin A/C