title: Methylation and gene expression responses to ethanol feeding and betaine supplementation in the cystathionine beta synthase-deficient mouse.
abstract: BACKGROUND
  Alcoholic steatohepatitis (ASH) is caused in part by the effects of ethanol (EtOH) on hepatic methionine metabolism.
  
  
  METHODS
  To investigate the phenotypic and epigenetic consequences of altered methionine metabolism in this disease, we studied the effects of 4-week intragastric EtOH feeding with and without the methyl donor betaine in cystathionine beta synthase (CβS) heterozygous C57BL/6J mice.
  
  
  RESULTS
  The histopathology of early ASH was induced by EtOH feeding and prevented by betaine supplementation, while EtOH feeding reduced and betaine supplementation maintained the hepatic methylation ratio of the universal methyl donor S-adenosylmethionine (SAM) to the methyltransferase inhibitor S-adenosylhomocysteine (SAH). MethylC-seq genomic sequencing of heterozygous liver samples from each diet group found 2 to 4% reduced methylation in gene bodies, but not promoter regions of all autosomes of EtOH-fed mice, each of which were normalized in samples from mice fed the betaine-supplemented diet. The transcript levels of nitric oxide synthase (Nos2) and DNA methyltransferase 1 (Dnmt1) were increased, while those of peroxisome proliferator receptor-α (Pparα) were reduced in EtOH-fed mice, and each was normalized in mice fed the betaine-supplemented diet. DNA pyrosequencing of CβS heterozygous samples found reduced methylation in a gene body of Nos2 by EtOH feeding that was restored by betaine supplementation and was correlated inversely with its expression and positively with SAM/SAH ratios.
  
  
  CONCLUSIONS
  The present study has demonstrated relationships among EtOH induction of ASH with aberrant methionine metabolism that was associated with gene body DNA hypomethylation in all autosomes and was prevented by betaine supplementation. The data imply that EtOH-induced changes in selected gene transcript levels and hypomethylation in gene bodies during the induction of ASH are a result of altered methionine metabolism that can be reversed through dietary supplementation of methyl donors.
authors.affiliation: Department of Internal Medicine, University of California Davis, Sacramento, California.
authors.forename: Valentina
authors.initials: V
authors.lastname: Medici
authors.forename: Diane I
authors.initials: DI
authors.lastname: Schroeder
authors.forename: Rima
authors.initials: R
authors.lastname: Woods
authors.forename: Janine M
authors.initials: JM
authors.lastname: LaSalle
authors.forename: Yongzhi
authors.initials: Y
authors.lastname: Geng
authors.forename: Noreene M
authors.initials: NM
authors.lastname: Shibata
authors.forename: Janet
authors.initials: J
authors.lastname: Peerson
authors.forename: Emir
authors.initials: E
authors.lastname: Hodzic
authors.forename: Sanjana
authors.initials: S
authors.lastname: Dayal
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Tsukamoto
authors.forename: Kusum K
authors.initials: KK
authors.lastname: Kharbanda
authors.forename: Brittany
authors.initials: B
authors.lastname: Tillman
authors.forename: Samuel W
authors.initials: SW
authors.lastname: French
authors.forename: Charles H
authors.initials: CH
authors.lastname: Halsted
mesh_terms: D000818:Animals; D001622:Betaine / Q000627:therapeutic use*; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000032:analysis; D019175:DNA Methylation / Q000187:drug effects*; D019587:Dietary Supplements; D000431:Ethanol / Q000494:pharmacology*; D005235:Fatty Liver, Alcoholic / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006712:Homocystinuria / Q000378:metabolism*; D008099:Liver / Q000737:chemistry / Q000187:drug effects; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D052247:Nitric Oxide Synthase Type II / Q000032:analysis; D047493:PPAR alpha / Q000032:analysis; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism
chemical_list: D047493:PPAR alpha; D000431:Ethanol; D001622:Betaine; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; D008715:Methionine; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619381:Dnmt1 protein, mouse; C000619382:Dnmt1 protein, rat
keywords: Alcohol; Cystathionine Beta Synthase; DNA Methylation; S-Adenosylhomocysteine; S-Adenosylmethionine
references.citation: Exp Mol Pathol. 2013 Feb;94(1):243-6
references.pmid: 23047067
references.citation: J Biol Chem. 2004 Nov 5;279(45):46954-61
references.pmid: 15308624
references.citation: Hepatology. 2004 May;39(5):1303-10
references.pmid: 15122759
references.citation: Biochem Pharmacol. 2005 Dec 5;70(12):1883-90
references.pmid: 16253211
references.citation: Science. 2010 May 14;328(5980):916-9
references.pmid: 20395474
references.citation: PLoS One. 2012;7(8):e41949
references.pmid: 22905112
references.citation: J Cell Mol Med. 2010 Jun;14(6A):1225-40
references.pmid: 20132414
references.citation: Nat Biotechnol. 2009 Apr;27(4):361-8
references.pmid: 19329998
references.citation: Can J Gastroenterol. 2000 Apr;14(4):327-32
references.pmid: 10799086
references.citation: Hepatology. 2010 Mar;51(3):932-41
references.pmid: 19957376
references.citation: Hum Mol Genet. 2011 Feb 15;20(4):670-80
references.pmid: 21112978
references.citation: Alcohol Clin Exp Res. 2007 Jul;31(7):1231-9
references.pmid: 17577393
references.citation: Nature. 2000 May 25;405(6785):421-4
references.pmid: 10839530
references.citation: Adv Nutr. 2011 Sep;2(5):421-7
references.pmid: 22332083
references.citation: Clin Chem. 2009 Apr;55(4):611-22
references.pmid: 19246619
references.citation: Alcohol Clin Exp Res. 2008 Sep;32(9):1525-34
references.pmid: 18616668
references.citation: Gastroenterology. 2003 May;124(5):1488-99
references.pmid: 12730887
references.citation: Alcohol Clin Exp Res. 2007 Nov;31(11):1934-43
references.pmid: 17850216
references.citation: Hepatology. 2004 Sep;40(3):565-73
references.pmid: 15349894
references.citation: Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1585-9
references.pmid: 7878023
references.citation: Blood. 2006 Oct 1;108(7):2237-43
references.pmid: 16804115
references.citation: BMC Bioinformatics. 2010 Apr 23;11:203
references.pmid: 20416082
references.citation: J Intern Med. 2007 May;261(5):488-99
references.pmid: 17444888
references.citation: Hepatology. 2013 Feb;57(2):555-65
references.pmid: 22945834
references.citation: Oncotarget. 2012 Apr;3(4):462-74
references.pmid: 22577155
references.citation: Nucleic Acids Res. 2008 Oct;36(18):e119
references.pmid: 18718928
references.citation: World J Gastroenterol. 2011 May 28;17(20):2456-64
references.pmid: 21633650
references.citation: Annu Rev Pharmacol Toxicol. 2001;41:203-36
references.pmid: 11264456
references.citation: World J Gastroenterol. 2006 Apr 21;12(15):2375-81
references.pmid: 16688828
references.citation: Semin Liver Dis. 2009 May;29(2):155-65
references.pmid: 19387915
references.citation: Proc Natl Acad Sci U S A. 2013 Apr 9;110(15):6037-42
references.pmid: 23530188
references.citation: Gastroenterology. 2012 Apr;142(4):824-833.e7
references.pmid: 22248660
references.citation: J Virol Methods. 2012 Feb;179(2):303-10
references.pmid: 22115785
references.citation: Am J Physiol Gastrointest Liver Physiol. 2000 Jul;279(1):G178-85
references.pmid: 10898761
references.citation: Nature. 2009 Nov 19;462(7271):315-22
references.pmid: 19829295
references.citation: Exp Mol Pathol. 2008 Apr;84(2):113-21
references.pmid: 18281034
references.citation: Gastroenterology. 2003 Dec;125(6):1834-44
references.pmid: 14724835
references.citation: Semin Liver Dis. 1993 May;13(2):154-69
references.pmid: 8393214
references.citation: J Biol Chem. 1998 Mar 6;273(10):5678-84
references.pmid: 9488698
references.citation: Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):10072-7
references.pmid: 12122204
references.citation: Hum Mol Genet. 2012 Jun 1;21(11):2399-411
references.pmid: 22343140
references.citation: Methods Mol Biol. 2008;447:33-48
references.pmid: 18369909
references.citation: J Biol Chem. 2002 Oct 4;277(40):37254-9
references.pmid: 12118000
references.citation: Int J Hepatol. 2012;2012:962183
references.pmid: 22187660
references.citation: J Hepatol. 2011 Sep;55(3):673-682
references.pmid: 21256905
references.citation: Am J Physiol Gastrointest Liver Physiol. 2005 Jul;289(1):G54-63
references.pmid: 15705656
references.citation: Genome Res. 2011 Oct;21(10):1583-91
references.pmid: 21784875
references.citation: Hepatology. 2007 May;45(5):1306-12
references.pmid: 17464973