We analyzed 27 578 CpG sites that map to 14 495 genes in omental arteries of normal pregnant and preeclamptic women for DNA methylation status using the Illumina platform. We found 1685 genes with a significant difference in DNA methylation at a false discovery rate of <10% with many inflammatory genes having reduced methylation. Unsupervised hierarchical clustering revealed natural clustering by diagnosis and methylation status. Of the genes with significant methylation differences, 236 were significant at a false discovery rate of <5%. When data were analyzed more stringently to a false discovery rate of <5% and difference in methylation of >0.10, 65 genes were identified, all of which showed reduced methylation in preeclampsia. When these genes were mapped to gene ontology for molecular functions and biological processes, 75 molecular functions and 149 biological processes were overrepresented in the preeclamptic vessels. These included smooth muscle contraction, thrombosis, inflammation, redox homeostasis, sugar metabolism, and amino acid metabolism. We speculate that reduced methylation may contribute to the pathogenesis of preeclampsia and that alterations in DNA methylation resulting from preeclampsia may increase maternal risk of cardiovascular disease later in life.
PridjianGPuschettJB. Preeclampsia. Part 2: experimental and genetic considerations. Obstet Gynecol Surv. 2002;57(9):619–640.
4.
WilliamsPJPipkinFB. The genetics of pre-eclampsia and other hypertensive disorders of pregnancy. Best practice & research. Clin obstet Gynaecol. 2011;25(4):405–417.
5.
WilsonCBMakarKWShnyrevaMFitzpatrickDR. DNA methylation and the expanding epigenetics of T cell lineage commitment. Semin Immunol. 2005;17(2):105–119.
6.
FragaMFEstellerM. Epigenetics and aging: the targets and the marks. Trends Genet. 2007;23(8):413–418.
7.
TostJ. DNA methylation: an introduction to the biology and the disease-associated changes of a promising biomarker. Mol Biotechnol. 2010;44(1):71–81.
8.
FrancoRSchoneveldOGeorgakilasAGPanayiotidisMI. Oxidative stress, DNA methylation and carcinogenesis. Cancer Lett. 2008;266(1):6–11.
9.
ZainaSLindholmMWLundG. Nutrition and aberrant DNA methylation patterns in atherosclerosis: more than just hyperhomocysteinemia?J Nutr. 2005;135(1):5–8.
10.
PaulsenMFerguson-SmithAC. DNA methylation in genomic imprinting, development, and disease. J Pathol. 2001;195(1):97–110.
11.
AgrawalAMurphyRFAgrawalDK. DNA methylation in breast and colorectal cancers. Mod Pathol. 2007;20(7):711–721.
12.
ConnorCMAkbarianS. DNA methylation changes in schizophrenia and bipolar disorder. Epigenetics. 2008;3:55–58.
13.
van der LindenIJHeilSGvan Egmont PetersenMvan StraatenHWden HeijerMBlomHJ. Inhibition of methylation and changes in gene expression in relation to neural tube defects. Birth Defects Res A Clin Mol Teratol. 2008;82(10):676–683.
14.
DongCYoonWGoldschmidt-ClermontPJ.DNA methylation and atherosclerosis. J Nutr. 2002;132:2406S–2409 S.
15.
BaccarelliAWrightRBollatiVLitonjuaAZanobettiATarantiniLSparrowDVokonasPSchwartzJ. Ischemic heart disease and stroke in relation to blood DNA methylation. Epidemiology. 2010;21(6):819–828.
16.
HubelCA. Oxidative stress in the pathogenesis of preeclampsia. Proc Soc Exp Biol Med. 1999;222(3):222–235.
17.
WalshSW. Maternal-placental interactions of oxidative stress and antioxidants in preeclampsia. Semin Reprod Endocrinol. 1998;16(1):93–104.
18.
WalshSW. Obesity: a risk factor for preeclampsia. Trends in endocrinology and metabolism: TEM. 2007;18(10):365–370.
19.
WangJXKnottnerusAMSchuitGNormanRJChanADekkerGA. Surgically obtained sperm, and risk of gestational hypertension and pre-eclampsia. Lancet. 2002;359(9307):673–674.
20.
YuLChenMZhaoDThe H19 gene imprinting in normal pregnancy and pre-eclampsia. Placenta. 2009;30(5):443–447.
21.
ChelbiSTVaimanD. Genetic and epigenetic factors contribute to the onset of preeclampsia. Mol Cell Endocrinol. 2008;282(1-2):120–129.
22.
ChelbiSTMondonFJammesHExpressional and epigenetic alterations of placental serine protease inhibitors: SERPINA3 is a potential marker of preeclampsia. Hypertension. 2007;49(1):76–83.
23.
KanayamaNTakahashiKMatsuuraTDeficiency in p57Kip2 expression induces preeclampsia-like symptoms in mice. Mol Hum Reprod. 2002;8(12):1129–1135.
24.
YuenRKPenaherreraMSvon DadelszenPMcFaddenDERobinsonWP. DNA methylation profiling of human placentas reveals promoter hypomethylation of multiple genes in early-onset preeclampsia. Eur J Hum Genet. 2010;18(9):1006–1012.
25.
ClarkSJStathamAStirzakerCMolloyPLFrommerM. DNA methylation: bisulphite modification and analysis. Nat Protoc. 2006;1:2353–2364.
XiongZLairdPW. COBRA: a sensitive and quantitative DNA methylation assay. Nucleic Acids Res. 1997;25(12):2532–2534.
28.
WangHOgawaMWoodJRGenetic and epigenetic mechanisms combine to control MMP1 expression and its association with preterm premature rupture of membranes. Hum Mol Genet. 2008;17(8):1087–1096.
29.
BibikovaMLinZZhouLHigh-throughput DNA methylation profiling using universal bead arrays. Genome Res. 2006;16(3):383–393.
30.
StoreyJDTibshiraniR. Statistical significance for genomewide studies. Proc Natl Acad Sci U S A. 2003;100(16):9440–9445.
31.
WeisenbergerDJBergDVD.PanFBermanBPLairdPW. Comprehensive DNA Methylation Analysis on the Illumina Infinium Assay Platform. San Diego, CA: Illumina and Inc; 2008.
32.
LiBCareyMWorkmanJL. The role of chromatin during transcription. Cell. 2007;128(4):707–719.
33.
ArcherKJMasVRMalufDGFisherRA. High-throughput assessment of CpG site methylation for distinguishing between HCV-cirrhosis and HCV-associated hepatocellular carcinoma. Mol Genet Genomics. 2010;283(4):341–349.
WalshSW. Preeclampsia: an imbalance in placental prostacyclin and thromboxane production. Am J Obstet Gynecol. 1985;152(3):335–340.
36.
ChavarriaMELara-GonzalezLGonzalez-GleasonAGarcia-PaletaYVital-ReyesVSReyesA. Prostacyclin/thromboxane early changes in pregnancies that are complicated by preeclampsia. Am J Obstet Gynecol. 2003;188(4):986–992.
37.
MillsJLDerSimonianRRaymondEProstacyclin and thromboxane changes predating clinical onset of preeclampsia: a multicenter prospective study. JAMA. 1999;282(4):356–362.
38.
WalshSW. Eicosanoids in preeclampsia. Prostaglandins Leukot Essent Fatty Acids. 2004;70:223–232.
39.
MousaAAStraussIII JFWalshSW. Reduced methylation of thromboxane synthase gene is correlated with its increased vascular expression in preeclampsia. Hypertension. 2012;59(6):1249–1255.
40.
Estrada-GutierrezGCappelloREMishraNRomeroRStraussJF3rd WalshSW. Increased expression of matrix metalloproteinase-1 in systemic vessels of preeclamptic women: a critical mediator of vascular dysfunction. Am J Pathol. 2011;178(1):451–460.
41.
CappelloREstrada-GutierrezGGerkPMStraussIII JFWalshSW.Epigenetic control of collagen regulating genes in vascular smooth muscle. Reprod Sci. 2008;15(supplement):73A.
42.
RedmanCWSacksGPSargentIL. Preeclampsia: an excessive maternal inflammatory response to pregnancy. Am J Obstet Gynecol. 1999;180(2 pt 1):499–506.
43.
GattiLTenconiPMGuarneriDHemostatic parameters and platelet activation by flow-cytometry in normal pregnancy: a longitudinal study. Int J Clin Lab Res. 1994;24(4):217–219.
44.
HalliganABonnarJSheppardBDarlingMWalsheJ. Haemostatic, fibrinolytic and endothelial variables in normal pregnancies and pre-eclampsia. Br J Obstet Gynaecol. 1994;101(6):488–492.
45.
HaramKAugensenKElsayedS. Serum protein pattern in normal pregnancy with special reference to acute-phase reactants. Br J Obstet Gynaecol. 1983;90(2):139–145.
46.
AustgulenRLienELiabakkNBJacobsenGArntzenKJ. Increased levels of cytokines and cytokine activity modifiers in normal pregnancy. Eur J Obstet Gynecol Reprod Biol. 1994;57(3):149–155.
47.
MelczerZBanhidyFCsomorSInfluence of leptin and the TNF system on insulin resistance in pregnancy and their effect on anthropometric parameters of newborns. Acta Obstet Gynecol Scand. 2003;82(5):432–438.
48.
ArntzenKJLiabakkNBJacobsenGEspevikTAustgulenR. Soluble tumor necrosis factor receptor in serum and urine throughout normal pregnancy and at delivery. Am J Reprod Immunol. 1995;34(3):163–169.
49.
LurieSRahamimEPiperIGolanASadanO. Total and differential leukocyte counts percentiles in normal pregnancy. Eur J Obstet Gynecol Reprod Biol. 2008;136(1):16–19.
50.
BarrigaCRodriguezABOrtegaE. Increased phagocytic activity of polymorphonuclear leukocytes during pregnancy. Eur J Obstet Gynecol Reprod Biol. 1994;57(1):43–46.
51.
SmarasonAKGunnarssonAAlfredssonJHValdimarssonH. Monocytosis and monocytic infiltration of decidua in early pregnancy. J Clin Lab Immunol. 1986;21(1):1–5.
52.
SacksGPStudenaKSargentKRedmanCW. Normal pregnancy and preeclampsia both produce inflammatory changes in peripheral blood leukocytes akin to those of sepsis. Am J Obstet Gynecol. 1998;179(1):80–86.
53.
BardenA. Circulating markers of oxidative stress are raised in normal pregnancy and pre-eclampsia. Br J Obstet Gynaecol. 1999;106(11):1232.
54.
MartinUDaviesCHayaviSHartlandADunneF. Is normal pregnancy atherogenic?Clin Sci (Lond). 1999;96(4):421–425.
55.
BorzychowskiAMSargentILRedmanCW. Inflammation and pre-eclampsia. Semin Fetal Neonatal Med. 2006;11(5):309–316.
56.
PlutzkyJ. The PPAR-RXR transcriptional complex in the vasculature: energy in the balance. Circ Res. 2011;108(8):1002–1016.
57.
ShahTJWalshSW.Activation of NF-kappaB and expression of COX-2 in association with neutrophil infiltration in systemic vascular tissue of women with preeclampsia. Am J Obstet Gynecol. 2007;196(1):48 e1–8.
58.
LotzeMTTraceyKJ. High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat Rev Immunol. 2005;5(4):331–342.
59.
YuMWangHDingAHMGB1 signals through toll-like receptor (TLR) 4 and TLR2. Shock. 2006;26(2):174–179.
60.
InoueKKawaharaKBiswasKKHMGB1 expression by activated vascular smooth muscle cells in advanced human atherosclerosis plaques. Cardiovasc Pathol. 2007;16(3):136–143.
61.
MihuDCostinNMihuCMBlagaLDPopRB. C-reactive protein, marker for evaluation of systemic inflammatory response in preeclampsia. Rev Med Chir Soc Med Nat Iasi. 2008;112(4):1019–1025.
62.
QiuCLuthyDAZhangCWalshSWLeisenringWMWilliamsMA. A prospective study of maternal serum C-reactive protein concentrations and risk of preeclampsia. Am J Hypertens. 2004;17(2):154–160.
63.
FiuzaCBustinMTalwarSTropeaMGerstenbergerEShelhamerJHSuffrediniAF. Inflammation-promoting activity of HMGB1 on human microvascular endothelial cells. Blood. 2003;101(7):2652–2660.
64.
LeikCEWalshSW. Neutrophils infiltrate resistance-sized vessels of subcutaneous fat in women with preeclampsia. Hypertension. 2004;44:72–77.
65.
NarumiyaHZhangYFernandez-PatronCGuilbertLJDavidgeST. Matrix metalloproteinase-2 is elevated in the plasma of women with preeclampsia. Hypertens Pregnancy. 2001;20(2):185–194.
KhalilRAGrangerJP. Vascular mechanisms of increased arterial pressure in preeclampsia: lessons from animal models. Am J Physiol Regul Integr Comp Physiol. 2002;283(1):R29–45.
68.
LunellNONylundLELewanderRSarbyB. Uteroplacental blood flow in pre-eclampsia measurements with indium-113m and a computer-linked gamma camera. Clin Exp Hypertens B. 1982;1(1):105–117.
69.
PerryKG.Jr., MartinJN,Jr. Abnormal hemostasis and coagulopathy in preeclampsia and eclampsia. Clin Obstet Gynecol. 1992;35(2):338–350.
70.
von Versen-HoeynckFMPowersRW. Maternal-fetal metabolism in normal pregnancy and preeclampsia. Front Biosci. 2007;12:2457–2470.
71.
HillLDYorkTPKusanovicJPEpistasis between COMT and MTHFR in maternal-fetal dyads increases risk for preeclampsia. PloS one. 2011;6(1):e16681.
72.
HitchlerMJDomannFE. An epigenetic perspective on the free radical theory of development. Free Radic Biol Med. 2007;43(7):1023–1036.
73.
WeitzmanSATurkPWMilkowskiDHKozlowskiK. Free radical adducts induce alterations in DNA cytosine methylation. Proc Natl Acad Sci U S A. 1994;91(4):1261–1264.
74.
MishraNNugentWHMahavadiSWalshSW. Mechanisms of enhanced vascular reactivity in preeclampsia. Hypertension. 2011;58(5):867–873.
75.
BellamyLCasasJPHingoraniADWilliamsDJ. Pre-eclampsia and risk of cardiovascular disease and cancer in later life: systematic review and meta-analysis. BMJ. 2007;335(7627):974.
76.
WenSWChenXKRodgerMFolic acid supplementation in early second trimester and the risk of preeclampsia. Am J Obstet Gynecol. 2008;198(1):45 e1–7.
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