Idiopathic male infertility is strongly associated with aberrant methylation of <i>MEST</i> and <i>IGF2/H19 ICR1</i>

<jats:title>Summary</jats:title><jats:p>Aberrant imprinting in spermatozoa in a subset of infertile men has been postulated to be a risk factor for congenital diseases in children conceived via assisted reproduction techniques (ART). Studies in clinically well characterized large cohorts, however, have been missing. Using bisulfite sequencing, we determined the degree of methylation of the <jats:italic>IGF2/H19</jats:italic> imprinting control region 1 (ICR1) and <jats:italic>MEST</jats:italic> differentially methylated regions in swim‐up purified spermatozoa from 148 idiopathic infertile men and 33 normozoospermic controls. All control individuals had a high degree of <jats:italic>IGF2/H19</jats:italic> ICR1 and a low degree of <jats:italic>MEST</jats:italic> methylation. Low sperm counts were clearly associated with <jats:italic>IGF2/H19</jats:italic> ICR1 hypomethylation and, even stronger, with <jats:italic>MEST</jats:italic> hypermethylation. <jats:italic>MEST</jats:italic> hypermethylation, but not <jats:italic>IGF2/H19</jats:italic> ICR1 hypomethylation was found in idiopathic infertile men with progressive sperm motility below 40% and bad sperm morphology below 5% normal spermatozoa. Ageing could be ruled out as a cause for the observed methylation defects. Sequence analysis of the <jats:italic>CTCFL</jats:italic> gene in peripheral blood DNA from 20 men with severe methylation defects revealed several polymorphisms, but no <jats:italic>bona fide</jats:italic> mutation. We conclude that idiopathic male infertility is strongly associated with imprinting defects at <jats:italic>IGF2/H19</jats:italic> ICR1 and <jats:italic>MEST</jats:italic>, with aberrant <jats:italic>MEST</jats:italic> methylation being a strong indicator for sperm quality. The male germ cell thus represents a potential source for aberrant epigenetic features in children conceived via ART.</jats:p>