Abstract
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.
Similar content being viewed by others
References
Cunningham FG, Leveno KJ, Bloom SL, Gilstrap III LC, Hauth JC, Wenstrom KD. Williams Obstetrics, 22nd ed. New York, NY: McGraw-Hill; 2005.
Roberts JM, Gammill HS. Preeclampsia: recent insights. Hypertension. 2005;46(6):1243–1249.
Pridjian G, Puschett JB. Preeclampsia. Part 2: experimental and genetic considerations. Obstet Gynecol Surv. 2002;57(9):619–640.
Williams PJ, Pipkin FB. The genetics of pre-eclampsia and other hypertensive disorders of pregnancy. Best practice & research. Clin obstet Gynaecol. 2011;25(4):405–417.
Wilson CB, Makar KW, Shnyreva M, Fitzpatrick DR. DNA methylation and the expanding epigenetics of T cell lineage commitment. Semin Immunol. 2005;17(2):105–119.
Fraga MF, Esteller M. Epigenetics and aging: the targets and the marks. Trends Genet. 2007;23(8):413–418.
Tost J. DNA methylation: an introduction to the biology and the disease-associated changes of a promising biomarker. Mol Biotechnol. 2010;44(1):71–81.
Franco R, Schoneveld O, Georgakilas AG, Panayiotidis MI. Oxidative stress, DNA methylation and carcinogenesis. Cancer Lett. 2008;266(1):6–11.
Zaina S, Lindholm MW, Lund G. Nutrition and aberrant DNA methylation patterns in atherosclerosis: more than just hyperhomocysteinemia? J Nutr. 2005;135(1):5–8.
Paulsen M, Ferguson-Smith AC. DNA methylation in genomic imprinting, development, and disease. J Pathol. 2001;195(1):97–110.
Agrawal A, Murphy RF, Agrawal DK. DNA methylation in breast and colorectal cancers. Mod Pathol. 2007;20(7):711–721.
Connor CM, Akbarian S. DNA methylation changes in schizophrenia and bipolar disorder. Epigenetics. 2008;3:55–58.
van der Linden IJ, Heil SG, van Egmont Petersen M, van Straaten HW, den Heijer M, Blom HJ. 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.
Dong C, Yoon W, Goldschmidt-Clermont PJ. DNA methylation and atherosclerosis. J Nutr. 2002;132:2406S-2409 S.
Baccarelli A, Wright R, Bollati V, Litonjua A, Zanobetti A, Tarantini L, Sparrow D, Vokonas P, Schwartz J. Ischemic heart disease and stroke in relation to blood DNA methylation. Epidemiology. 2010;21(6):819–828.
Hubel CA. Oxidative stress in the pathogenesis of preeclampsia. Proc Soc Exp Biol Med. 1999;222(3):222–235.
Walsh SW. Maternal-placental interactions of oxidative stress and antioxidants in preeclampsia. Semin Reprod Endocrinol. 1998; 16(1):93–104.
Walsh SW. Obesity: a risk factor for preeclampsia. Trends in endocrinology and metabolism: TEM. 2007;18(10):365–370.
Wang JX, Knottnerus AM, Schuit G, Norman RJ, Chan A, Dekker GA. Surgically obtained sperm, and risk of gestational hypertension and pre-eclampsia. Lancet. 2002;359(9307):673–674.
Yu L, Chen M, Zhao D, et al. The H19 gene imprinting in normal pregnancy and pre-eclampsia. Placenta. 2009;30(5):443–447.
Chelbi ST, Vaiman D. Genetic and epigenetic factors contribute to the onset of preeclampsia. Mol Cell Endocrinol. 2008;282(1–2): 120–129.
Chelbi ST, Mondon F, Jammes H, et al. Expressional and epigenetic alterations of placental serine protease inhibitors: SERPINA3 is a potential marker of preeclampsia. Hypertension. 2007;49(1):76–83.
Kanayama N, Takahashi K, Matsuura T, et al. Deficiency in p57Kip2 expression induces preeclampsia-like symptoms in mice. Mol Hum Reprod. 2002;8(12):1129–1135.
Yuen RK, Penaherrera MS, von Dadelszen P, McFadden DE, Robinson WP. DNA methylation profiling of human placentas reveals promoter hypomethylation of multiple genes in earlyonset preeclampsia. Eur J Hum Genet. 2010;18(9):1006–1012.
Clark SJ, Statham A, Stirzaker C, Molloy PL, Frommer M. DNA methylation: bisulphite modification and analysis. Nat Protoc. 2006;1:2353–2364.
Grunau C, Clark SJ, Rosenthal A. Bisulfite genomic sequencing: systematic investigation of critical experimental parameters. Nucleic Acids Res. 2001;29(13):E65–5.
Xiong Z, Laird PW. COBRA: a sensitive and quantitative DNA methylation assay. Nucleic Acids Res. 1997;25(12):2532–2534.
Wang H, Ogawa M, Wood JR, et al. Genetic 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.
Bibikova M, Lin Z, Zhou L, et al. High-throughput DNA methylation profiling using universal bead arrays. Genome Res. 2006; 16(3):383–393.
Storey JD, Tibshirani R. Statistical significance for genomewide studies. Proc Natl Acad Sci U S A. 2003;100(16):9440–9445.
Weisenberger DJ, Berg DVD., Pan F, Berman BP, Laird PW. Comprehensive DNA Methylation Analysis on the Illumina Infinium Assay Platform. San Diego, CA: Illumina and Inc; 2008.
Li B, Carey M, Workman JL. The role of chromatin during transcription. Cell. 2007;128(4):707–719.
Archer KJ, Mas VR, Maluf DG, Fisher RA. 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.
Tanabe T, Ullrich V. Prostacyclin and thromboxane synthases. J Lipid Mediat Cell Signal. 1995;12:243–255.
Walsh SW. Preeclampsia: an imbalance in placental prostacyclin and thromboxane production. Am J Obstet Gynecol. 1985;152(3): 335–340.
Chavarria ME, Lara-Gonzalez L, Gonzalez-Gleason A, Garcia-Paleta Y, Vital-Reyes VS, Reyes A. Prostacyclin/thromboxane early changes in pregnancies that are complicated by preeclampsia. Am J Obstet Gynecol. 2003;188(4):986–992.
Mills JL, DerSimonian R, Raymond E, et al. Prostacyclin and thromboxane changes predating clinical onset of preeclampsia: a multicenter prospective study. JAMA. 1999;282(4):356–362.
Walsh SW. Eicosanoids in preeclampsia. Prostaglandins Leukot Essent Fatty Acids. 2004;70:223–232.
Mousa AA, Strauss III JF, Walsh SW. Reduced methylation of thromboxane synthase gene is correlated with its increased vascular expression in preeclampsia. Hypertension. 2012;59(6): 1249–1255.
Estrada-Gutierrez G, Cappello RE, Mishra N, Romero R, Strauss JF 3rd Walsh SW. 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.
Cappello R, Estrada-Gutierrez G, Gerk PM, Strauss III JF, Walsh SW. Epigenetic control of collagen regulating genes in vascular smooth muscle. Reprod Sci. 2008;15(supplement):73A.
Redman CW, Sacks GP, Sargent IL. Preeclampsia: an excessive maternal inflammatory response to pregnancy. Am J Obstet Gynecol. 1999;180(2 pt 1):499–506.
Gatti L, Tenconi PM, Guarneri D, et al. Hemostatic parameters and platelet activation by flow-cytometry in normal pregnancy: a longitudinal study. Int J Clin Lab Res. 1994;24(4): 217–219.
Halligan A, Bonnar J, Sheppard B, Darling M, Walshe J. Haemostatic, fibrinolytic and endothelial variables in normal pregnancies and pre-eclampsia. Br J Obstet Gynaecol. 1994; 101(6):488–492.
Haram K, Augensen K, Elsayed S. Serum protein pattern in normal pregnancy with special reference to acute-phase reactants. Br J Obstet Gynaecol. 1983;90(2):139–145.
Austgulen R, Lien E, Liabakk NB, Jacobsen G, Arntzen KJ. Increased levels of cytokines and cytokine activity modifiers in normal pregnancy. Eur J Obstet Gynecol Reprod Biol. 1994; 57(3):149–155.
Melczer Z, Banhidy F, Csomor S, et al. Influence 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.
Arntzen KJ, Liabakk NB, Jacobsen G, Espevik T, Austgulen R. Soluble tumor necrosis factor receptor in serum and urine throughout normal pregnancy and at delivery. Am J Reprod Immunol. 1995;34(3):163–169.
Lurie S, Rahamim E, Piper I, Golan A, Sadan O. Total and differential leukocyte counts percentiles in normal pregnancy. Eur J Obstet Gynecol Reprod Biol. 2008;136(1):16–19.
Barriga C, Rodriguez AB, Ortega E. Increased phagocytic activity of polymorphonuclear leukocytes during pregnancy. Eur J Obstet Gynecol Reprod Biol. 1994;57(1):43–46.
Smarason AK, Gunnarsson A, Alfredsson JH, Valdimarsson H. Monocytosis and monocytic infiltration of decidua in early pregnancy. J Clin Lab Immunol. 1986;21(1):1–5.
Sacks GP, Studena K, Sargent K, Redman CW. 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.
Barden A. Circulating markers of oxidative stress are raised in normal pregnancy and pre-eclampsia. Br J Obstet Gynaecol. 1999;106(11):1232.
Martin U, Davies C, Hayavi S, Hartland A, Dunne F. Is normal pregnancy atherogenic? Clin Sci (Lond). 1999;96(4): 421–425.
Borzychowski AM, Sargent IL, Redman CW. Inflammation and pre-eclampsia. Semin Fetal Neonatal Med. 2006;11(5):309–316.
Plutzky J. The PPAR-RXR transcriptional complex in the vasculature: energy in the balance. Circ Res. 2011;108(8):1002–1016.
Shah TJ, Walsh SW. 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.
Lotze MT, Tracey KJ. High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat Rev Immunol. 2005;5(4):331–342.
Yu M, Wang H, Ding A, et al. HMGB1 signals through toll-like receptor (TLR) 4 and TLR2. Shock. 2006;26(2):174–179.
Inoue K, Kawahara K, Biswas KK, et al. HMGB1 expression by activated vascular smooth muscle cells in advanced human atherosclerosis plaques. Cardiovasc Pathol. 2007;16(3): 136–143.
Mihu D, Costin N, Mihu CM, Blaga LD, Pop RB. C-reactive protein, marker for evaluation of systemic inflammatory response in preeclampsia. Rev Med Chir Soc Med Nat Iasi. 2008;112(4): 1019–1025.
Qiu C, Luthy DA, Zhang C, Walsh SW, Leisenring WM, Williams MA. A prospective study of maternal serum C-reactive protein concentrations and risk of preeclampsia. Am J Hypertens. 2004;17(2):154–160.
Fiuza C, Bustin M, Talwar S, Tropea M, Gerstenberger E, Shelhamer JH, Suffredini AF. Inflammation-promoting activity of HMGB1 on human microvascular endothelial cells. Blood. 2003;101(7):2652–2660.
Leik CE, Walsh SW. Neutrophils infiltrate resistance-sized vessels of subcutaneous fat in women with preeclampsia. Hypertension. 2004;44:72–77.
Narumiya H, Zhang Y, Fernandez-Patron C, Guilbert LJ, Davidge ST. Matrix metalloproteinase-2 is elevated in the plasma of women with preeclampsia. Hypertens Pregnancy. 2001;20(2): 185–194.
Poon LC, Nekrasova E, Anastassopoulos P, Livanos P, Nicolaides KH. First-trimester maternal serum matrix metalloproteinase-9 (MMP-9) and adverse pregnancy outcome. Prenat Diagn. 2009; 29(6):553–559.
Khalil RA, Granger JP. Vascular mechanisms of increased arterial pressure in preeclampsia: lessons from animal models. Am J Physiol Regul Integr Comp Physiol. 2002;283(1):R29–45.
Lunell NO, Nylund LE, Lewander R, Sarby B. 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.
Perry KG. Jr., Martin JN, Jr. Abnormal hemostasis and coagulopathy in preeclampsia and eclampsia. Clin Obstet Gynecol. 1992;35(2):338–350.
von Versen-Hoeynck FM, Powers RW. Maternal-fetal metabolism in normal pregnancy and preeclampsia. Front Biosci. 2007;12: 2457–2470.
Hill LD, York TP, Kusanovic JP, et al. Epistasis between COMT and MTHFR in maternal-fetal dyads increases risk for preeclampsia. PloS one. 2011;6(1):e16681.
Hitchler MJ, Domann FE. An epigenetic perspective on the free radical theory of development. Free Radic Biol Med. 2007; 43(7):1023–1036.
Weitzman SA, Turk PW, Milkowski DH, Kozlowski K. Free radical adducts induce alterations in DNA cytosine methylation. Proc Natl Acad Sci U S A. 1994;91(4):1261–1264.
Mishra N, Nugent WH, Mahavadi S, Walsh SW. Mechanisms of enhanced vascular reactivity in preeclampsia. Hypertension. 2011;58(5):867–873.
Bellamy L, Casas JP, Hingorani AD, Williams DJ. Preeclampsia and risk of cardiovascular disease and cancer in later life: systematic review and meta-analysis. BMJ. 2007; 335(7627):974.
Wen SW, Chen XK, Rodger M, et al. Folic acid supplementation in early second trimester and the risk of preeclampsia. Am J Obstet Gynecol. 2008;198(1):45 e1–7.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mousa, A.A., Archer, K.J., Cappello, R. et al. DNA Methylation is Altered in Maternal Blood Vessels of Women With Preeclampsia. Reprod. Sci. 19, 1332–1342 (2012). https://doi.org/10.1177/1933719112450336
Published:
Issue Date:
DOI: https://doi.org/10.1177/1933719112450336