advanced paternal age blastocyst DNA methylation epigenetics offspring neurodevelopmental disorders sperm A statistical comparison of the sperm and blastocyst DMR‐associated genes demonstrated a highly significant enrichment of genes between the two methylomes upon paternal aging ( p = 3.46 × 10 ‐55 ; odds ratio [OR] 3.19).
Pyrosequencing data confirmed significant hypomethylation along the amplified CACNA1H DMR region, with 40% average methylation for the young paternal age blastocyst group significantly decreased to 25% in the APA blastocyst group ( p < .05), while validation of the SHANK2 blastocyst DMR showed a trend toward hypomethylation (young: 75%, APA: 56%; p < .1) (Figure S2 ). 2.3 Localization of DMRs To determine whether specific chromosomal regions were more susceptible to age‐related methylation alterations, we analyzed chromosomal enrichment for DMR‐associated gene density at individual cytobands ( p < .05, q < 0.05).