Promoter targeted bisulfite sequencing reveals DNA methylation profiles associated with low sperm motility in asthenozoospermia
Promoter targeted bisulfite sequencing reveals DNA methylation profiles associated with low sperm motility in asthenozoospermia
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启动子靶向亚硫酸氢盐测序揭示了与弱精子症低精子活力相关的 DNA 甲基化谱
DOI：
10.1093/humrep/dev283
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发表时间：
2016-01-01
影响因子：
6.1
通讯作者：
Jiang, Hui
中科院分区：
文献类型：
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作者：
Du, Ye;Li, Meiyan;Jiang, Hui
研究问题：精子DNA甲基化谱与弱精子症之间是否存在关联？总结答复：DNA甲基化，在特定的CpG而不是在整体水平上，在弱精子症个体的低活动性精子细胞和正常精子症对照的高活动性精子细胞之间存在显著差异。已知：DNA甲基化异常，无论是整体的还是局限于特定基因位点的，都与男性不育和精液参数异常有关。这是一项病例对照研究，调查8名正常精子症对照和7名弱精子症患者的高和低运动精子细胞启动子区CpG的DNA甲基化差异。使用基于液体杂交捕获的亚硫酸氢盐测序方法来确定启动子区域中CpG的DNA甲基化。通过评估来自相同或不同个体的不同活动精子组分之间和内部的甲基化方差，估计整体个体间和个体内甲基化变异性。通过比较正常精子对照组的高活动性精子细胞与弱精子症患者的低活动性精子细胞的DNA甲基化，确定弱精子症相关的差异甲基化或可变CpG和差异甲基化区域。在这项研究中，我们确定了整体DNA甲基化水平（24.7%），个体间方差（14.4%）和高和低活动精子分数之间的个体差异（3.9%）。我们证明了正常精子症和弱精子症男性精子的DNA甲基化水平和甲基化变异性在统计学上没有显著差异。在正常精子症男性的高活力精子和弱精子症男性的低活力精子之间，我们鉴定了134个差异甲基化的CpG，41个差异甲基化的区域和134个差异可变的CpG。差异甲基化谱的基因组分布模式表明，弱精子症患者低运动精子细胞的基因表达可能受到影响。最后，通过功能分析，我们检测到16个差异甲基化或可变的基因，所需的精子发生和精子活力或主要在testis.Limitations表达，理由回避：在这项研究中的样本量是有限的，虽然在两组的参与者都是经过精心挑选和匹配。我们的研究结果必须在更大的队列中使用不同的技术进行验证。此外，我们的研究结果是描述性的，后续的研究将需要阐明差异甲基化对弱精子症的影响。更广泛的影响的调查结果：我们的研究确定了弱精子症相关的DNA甲基化谱，并提出了一个列表的基因，这被认为是通过改变DNA甲基化参与调节精子活力。这些结果将为理解DNA甲基化对精子运动和弱精子症的影响提供有希望的线索。
STUDY QUESTION: Is there an association between sperm DNA methylation profiles and asthenozoospermia?SUMMARYANSWER: DNA methylation, at specific CpGs but not at the global level, was significantly different between low motile sperm cells of asthenozoospermic individuals and high motile sperm cells of normozoospermic controls.WHAT IS KNOWN ALREADY: Aberrant DNA methylation, both globally and restricted to a specific gene locus, has been associated with male infertility and abnormal semen parameters.STUDY DESIGN, SIZE, DURATION: This was a case-control study investigating the differences in DNA methylation at CpGs in promoter regions between high and low motile sperm cells from eight normozoospermic controls and seven asthenozoospermic patients.PARTICIPANTS/MATERIALS, SETTING, METHODS: The liquid hybridization capture-based bisulfite sequencing method was used to determine DNA methylation at CpGs in promoter regions. The global inter-individual and intra-individual methylation variability were estimated by evaluating the methylation variance between and within different motile sperm fractions from the same or different individuals. Asthenozoospermia-associated differentially methylated or variable CpGs and differentially methylated regions were identified by comparing the DNA methylation of high motile sperm cells from normozoospermic controls with that of low motile sperm cells from asthenozoospermic patients.MAIN RESULTS AND THE ROLE OF CHANCE: In this study, we determined the global DNA methylation level (24.7%), inter-individual variance (14.4%) and intra-individual differences between high and low motile sperm fractions (3.9%). We demonstrated that there were no statistically signiinot signcant differences in either the global DNA methylation level or global methylation variability between sperm from men with normozoospermia or asthenozoospermia. Between high motile sperm from men with normozoospermia and low motile sperm from men with asthenozoospermia, we identified 134 differentially methylated CpGs, 41 differentially methylated regions and 134 differentially variable CpGs. The genomic distribution patterns of the differential methylation spectrum suggested that gene expression may be affected in low motile sperm cells of asthenozoospermic patients. Finally, through a functional analysis, we detected 16 differentially methylated or variable genes that are required for spermatogenesis and sperm motility or dominantly expressed in testis.LIMITATIONS, REASONS FOR CAUTION: The sample size in this study was limited, although the participants in the two groups were carefully selected and well matched. Our results must be verified in larger cohorts with the use of different techniques. Furthermore, our results were descriptive, and follow-up studies will be needed to elucidate the effect of differential methylation profiles on asthenozoospermia.WIDER IMPLICATIONS OF THE FINDINGS: Our study identified asthenozoospermia-associated DNA methylation profiles and proposed a list of genes, which were suggested to be involved in the regulation of sperm motility through an alteration of DNA methylation. These results will provide promising clues for understanding the effect of DNA methylation on sperm motility and asthenozoospermia.