ABSTRACT

Clinical and experimental evidences indicate that epigenetic modifications induced by the prenatal environment are related to metabolic and reproductive derangements in polycystic ovary syndrome (PCOS). Alterations in the leptin and adiponectin systems, androgen signalling and antimüllerian hormone (AMH) levels have been observed in PCOS women and in their offspring. Using a targeted Next-Generation Sequencing (NGS), we studied DNA methylation in promoter regions of the leptin (LEP), leptin receptor (LEPR), adiponectin (ADIPOQ), adiponectin receptor 1 and 2 (ADIPOR1 and ADIPOR2), AMH and androgen receptor (AR) genes in 24 sons and daughters of women with PCOS (12 treated with metformin during pregnancy) and 24 children born to non-PCOS women during early infancy (2–3 months of age). Genomic DNA was extracted from whole blood, bisulphite converted and sequenced by NGS. Girls showed differences between groups in 1 CpG site of LEPR, 2 of LEP, 1 of ADIPOR2 and 2 of AR. Boys showed differences in 5 CpG sites of LEP, 3 of AMH and 9 of AR. Maternal metformin treatment prevented some of these changes in LEP, ADIPOR2 and partially in AR in girls, and in LEP and AMH in boys. Maternal BMI at early pregnancy was inversely correlated with the methylation levels of the ChrX-67544981 site in the whole group of girls (r = −0.530, p = 0.008) and with the global Z-score in all boys (r = −0.539, p = 0.007). These data indicate that the intrauterine PCOS environment predisposes the offspring to acquire certain sex-dependent DNA methylation patterns in the promoter regions of metabolic and reproductive genes.

摘要

临床和实验证据表明，产前环境引起的表观遗传修饰与多囊卵巢综合征（PCOS）的代谢和生殖紊乱有关。已在PCOS妇女及其后代中观察到瘦素和脂联素系统的变化，雄激素信号和抗苗勒管激素（AMH）的水平。使用针对性的下一代测序（NGS），我们研究了瘦素（LEP），瘦素受体（LEPR），脂联素（ADIPOQ），脂联素受体1和2（ADIPOR1和ADIPOR2），AMH和雄激素的启动子区域的DNA甲基化受体（AR）24例患有PCOS的妇女的儿女（在怀孕期间接受二甲双胍治疗的12个儿女）和非PCOS妇女在婴儿早期（2-3月龄）所生的24个孩子中的基因。从全血中提取基因组DNA，将亚硫酸氢盐转化并通过NGS测序。女孩在LEPR的1个CpG部位，LEP的2个部位，ADIPOR2的1个部位和AR的2个部位之间表现出差异。男孩在LEP的5个CpG位点，AMH的3个和AR的9个上有差异。产妇二甲双胍治疗可预防部分女孩的LEP，ADIPOR2和部分AR的这些变化，以及LEP和男孩的AMH。在整个女孩组中，孕妇早孕时的BMI与ChrX-67544981位点的甲基化水平呈负相关（r = -0.530，p = 0.008），与所有男孩的总体Z得分（r = -0.539， p = 0.007）。这些数据表明子宫内PCOS环境使后代易于在代谢和生殖基因的启动子区域中获得某些性别依赖性的DNA甲基化模式。