DNA methylation in spermatozoa as a prospective marker in andrology
DNA methylation in spermatozoa as a prospective marker in andrology
复制标题
DOI：
10.1111/j.2047-2927.2013.00118.x
复制
发表时间：
2013-09-01
期刊：
影响因子：
4.5
通讯作者：
Gromoll, J.
中科院分区：
文献类型：
--
作者：
Klaever, R.;Tuettelmann, F.;Gromoll, J.
最近的研究表明精子中异常的DNA甲基化与特发性不育有关。因此，特定基因的DNA甲基化分析可以作为临床男科诊断的一个有价值的标志物。为此，需要建立和验证精子DNA甲基化的快速可靠的检测方法、参考值和时间稳定性。在这项前瞻性研究中，收集了212例连续患者(单个样本)、31名正常精子志愿者(单个样本)和10名正常精子志愿者(第1、3、42、45天的4份样本和180-951天后的第5份样本)的上游精液样本。提取精子DNA，亚硫酸氢盐转化，用焦磷酸测序分析DNA甲基化。对212例患者和31例正常精子志愿者的母系印记基因MEST进行了DNA甲基化检测，并对10例正常精子志愿者连续采集的8个不同基因和2个重复序列的时间稳定性进行了检测。Mest DNA甲基化与少精子症、双睾丸体积缩小和FSH水平升高显著相关。在正常精子志愿者中，利用DNA甲基化的第95个百分位数建立了精子MEST DNA甲基化的参考范围(0-15%)。使用这个参考范围，我们的队列中大约23%的患者表现出异常的MEST DNA甲基化。这种表观遗传学异常被发现与双睾丸体积、精子密度和总精子数显著相关。与经典的精液参数相比，正常精子志愿者的DNA甲基化在长达951天的时间内是稳定的。我们的数据显示，MEST DNA甲基化满足用作常规参数的先决条件，并支持在男科检查中使用它，如果未来能显示预后价值的话。
Recent studies have shown associations of aberrant DNA methylation in spermatozoa with idiopathic infertility. The analysis of DNA methylation of specific genes could therefore serve as a valuable diagnostic marker in clinical andrology. For this purpose, rapid and reliable detection methods, reference values and the temporal stability of spermatozoal DNA methylation need to be established and demonstrated. In this prospective study, swim-up purified semen samples from 212 consecutive patients (single samples), 31 normozoospermic volunteers (single samples) and 10 normozoospermic volunteers (four samples at days 1, 3, 42 and 45 plus a fifth sample after 180-951 days) were collected. Spermatozoal DNA was isolated, bisulphite converted and DNA methylation was analysed by pyrosequencing. DNA methylation of the maternally imprinted gene MEST was measured in samples of 212 patients and 31 normozoospermic volunteers and the temporal stability of eight different genes and two repetitive elements was examined in consecutive samples of 10 normozoospermic volunteers. MEST DNA methylation was significantly associated with oligozoospermia, decreased bi-testicular volume and increased FSH levels. A reference range for spermatozoal MEST DNA methylation (0-15%) was established using the 95th percentile of DNA methylation in normozoospermic volunteers. Using this reference range, around 23% of our patient cohort displayed an aberrant MEST DNA methylation. This epigenetic aberration was found to be significantly associated with bi-testicular volume, sperm concentration and total sperm count. DNA methylation in normozoospermic volunteers was stable over a time period of up to 951 days in contrast to classical semen parameters. Our data show that MEST DNA methylation fulfils the prerequisites to be used as routine parameter and support its use during andrological workup if a prognostic value can be shown in future.