Quantitative methylation analysis of developmentally important genes in human pregnancy losses after ART and spontaneous conception.
To study possible effects of assisted reproductive technologies (ART) on epigenetic reprogramming, we have analyzed the DNA methylation levels of differentially methylated regions (DMRs) of seven imprinted genes (H19, MEG3, LIT1, MEST, NESP55, PEG3 and SNRPN) as well as the promoter regions of the pluripotency gene NANOG and the tumor suppressor gene APC in chorionic villus samples (CVS) of 42 spontaneous miscarriages and stillbirths after ART and 29 abortions/stillbirths after spontaneous conception. We did not find an increased rate of faulty methylation patterns after ART, but significant and trend differences (ROC curve analysis, Wilcoxon test) in the methylation levels of LIT1 (P = 0.006) and H19 (P = 0.085) between ART and non-ART samples. With the possible exception of NANOG, we did not observe a gestational age effect on the methylation levels of the studied genes. The frequency of extreme methylation values in PEG3 and APC was markedly higher than in the other studied genes, indicating an increased susceptibility of some genes to epigenetic alterations. Most methylation abnormalities in CVS represented either hypermethylated DMRs of paternally and maternally imprinted genes or hypomethylated promoters of non-imprinted genes. The observed methylation abnormalities (mosaicism) are consistent with methylation reprogramming defects during early embryogenesis.
Zechner
Ulrich
U
Institute of Human Genetics, Johannes Gutenberg University, 55101 Mainz, Germany.
Pliushch
Galyna
G
Schneider
Eberhard
E
El Hajj
Nady
N
Tresch
Achim
A
Shufaro
Yoel
Y
Seidmann
Larissa
L
Coerdt
Wiltrud
W
Müller
Annette M
AM
Haaf
Thomas
T
D000022:Abortion, Spontaneous / Q000235:genetics*; D000328:Adult; D019175:DNA Methylation; D005260:Female; D018507:Gene Expression Regulation, Developmental; D017491:Genes, APC; D020022:Genetic Predisposition to Disease; D005858:Germany; D005865:Gestational Age; D006801:Humans; D007557:Israel; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D016014:Linear Models; D008423:Maternal Age; D008875:Middle Aged; D009030:Mosaicism; D010641:Phenotype; D011247:Pregnancy; D027724:Reproductive Techniques, Assisted / Q000009:adverse effects*; D018570:Risk Assessment; D012307:Risk Factors; D050497:Stillbirth / Q000235:genetics*; D055815:Young Adult
D051741:Kruppel-Like Transcription Factors; C490820:PEG3 protein, human