Laqqan Mohammed, Solomayer Erich-Franz, Hammadeh Mohamed
Department of Obstetrics, Gynecology & Assisted Reproduction Laboratory, Saarland University, Homburg, Germany.
Department of Obstetrics, Gynecology & Assisted Reproduction Laboratory, Saarland University, Homburg, Germany.
Reprod Biol. 2017 Sep;17(3):246-251. doi: 10.1016/j.repbio.2017.05.010. Epub 2017 May 26.
Infertility affects about 15% of couples worldwide, with approximately 7% of males suffering from infertility problems. This study was designed to assess the relationship between alterations in sperm DNA methylation patterns and semen parameters in subfertile males. Of a total of 108 males, 30 samples were subjected to 450K BeadChip arrays to evaluate the variation in DNA methylation level between cases and controls. Three CpG sites showed the highest difference in methylation levels (cg09737095, cg14271023, and cg17662493), which are located in the KCNJ5, MLPH, and SMC1β genes, respectively; these were selected for further analysis using deep bisulfite sequencing in 78 independent samples (21 proven fertile "controls", and 57 subfertile "cases"). The results of a validation study showed that variation in methylation levels was found in more than one CpG site: there was a significant decrease in methylation levels at six CpGs (CpG1, CpG3, CpG4, CpG6, CpG7, and CpG8) in the KCNJ5 gene-related amplicon (p≤0.001, p≤0.009, p≤0.007, p≤0.007, p≤0.020, and p≤0.016, respectively), and at (CpG1, CpG2, and CpG4) in the MLPH gene-related amplicon (p≤0.003, p≤0.005, and p≤0.0001, respectively), while there was a significant increase in the methylation level at six out of eight CpGs in the SMC1β gene-related amplicon in cases compared to controls. Our results show that three CpGs have a significant difference in sperm DNA methylation levels in subfertile males compared to proven fertile males.
不孕症影响着全球约15%的夫妇，其中约7%的男性患有不孕问题。本研究旨在评估亚生育男性精子DNA甲基化模式改变与精液参数之间的关系。在总共108名男性中，30个样本接受了450K BeadChip阵列检测，以评估病例组和对照组之间DNA甲基化水平的差异。三个CpG位点的甲基化水平差异最大（cg09737095、cg14271023和cg17662493），它们分别位于KCNJ5、MLPH和SMC1β基因中；在78个独立样本（21名已证实可育的“对照”和57名亚生育的“病例”）中，选择这些位点使用深度亚硫酸氢盐测序进行进一步分析。验证研究结果表明，在多个CpG位点发现了甲基化水平的变化：在KCNJ5基因相关扩增子中，六个CpG（CpG1、CpG3、CpG4、CpG6、CpG7和CpG8）的甲基化水平显著降低（分别为p≤0.001、p≤0.009、p≤0.007、p≤0.007、p≤0.020和p≤0.016），在MLPH基因相关扩增子中（CpG1、CpG2和CpG4）的甲基化水平也显著降低（分别为p≤0.003、p≤0.005和p≤0.0001），而与对照组相比，病例组中SMC1β基因相关扩增子的八个CpG中有六个的甲基化水平显著升高。我们的结果表明，与已证实可育的男性相比，亚生育男性精子DNA甲基化水平在三个CpG位点存在显著差异。
