DNA methylation defects in spermatozoa of male partners from couples experiencing recurrent pregnancy loss
DNA methylation defects in spermatozoa of male partners from couples experiencing recurrent pregnancy loss
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DOI：
10.1093/humrep/deaa278
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发表时间：
2021-01-01
影响因子：
6.1
通讯作者：
Balasinor, Nafisa H.
中科院分区：
文献类型：
--
作者：
Khambata, Kushaan;Raut, Sanketa;Balasinor, Nafisa H.
研究问题：经历反复流产（RPL）的夫妇的男性伴侣精子DNA印迹基因甲基化状态如何？总结回答：胰岛素样生长因子-2-H19差异甲基化区域等印迹基因精子DNA甲基化状态的异常（IGF 2-H19 DMR）、基因间差异甲基化区域（IG-DMR）、中胚层特异性转录物（MEST）、调节凋亡和细胞周期停滞的锌指蛋白（ZAC）、KCNQ 1基因内含子10中的DMR（KvDMR）、父系表达基因3（PEG 3）和父系表达基因10（PEG 10），以及精子总体5-甲基胞嘧啶（5 mC）水平的降低，与RPL有关。虽然有几个母亲和父亲的病因学因素有助于RPL，近50%的情况下仍然是特发性的。因此，有必要确定推定的父亲因素，可能有助于对妊娠丢失的情况下，特发性RPL.Study设计，规模，持续时间：在这个病例对照研究中，112对夫妇接受RPL没有明确的原因，从2015年9月至2018年5月招募。对照组包括106名健康的被证明有生育能力的夫妇，没有不孕或流产的历史。参与者/材料，设置，方法：在这项研究中，我们调查了父亲的遗传和表观遗传因素，可能与RPL。我们研究了DNA甲基化，焦磷酸测序，选定的印记基因牵连在胚胎发育，如IGF 2-H19 DMR，IG-DMR，MEST，ZAC，KvDMR，PEG 3，PEG 10和小核核糖核蛋白多肽N（SNRPN）的男性精子的伴侣目前RPL。通过研究5 mC含量和长散布核元件I（LINEI）启动子甲基化来评估精子中的总体DNA甲基化。我们还通过焦磷酸测序研究了IGF 2 HI 9 DMR以及IGF 2启动子中的多态性。主要结果和机会的作用：在RPL组中，我们发现在LINE中的三个CpG位点处的全球精子5 mC水平和DNA甲基化的显著降低显著降低。启动子对于IGF 2 H19 DMR和IG，在RPL组中观察到精子特异性CpG位点的DNA甲基化显著降低。对于母系印记基因，如MEST、ZAC、KvDMR、PEG 3和PEG 10，注意到超甲基化。IGF 2 H19 DMR和IGF 2的多态性研究揭示了基因型频率在男性中的显着差异。模仿，预防的原因：在这项研究中，我们分析了选定的候选印迹基因的甲基化水平与胚胎发育有关。发生在全基因组水平的甲基化变化的检测可能会发现进一步的候选基因有一个更好的区别控制和研究groups. WIDER的影响的发现：我们的研究表明，某些多态性和异常精子甲基化状态的印记基因与RPL，可能有助于RPL的病因。这项研究表明，调查父亲的遗传和表观遗传因素可能是有用的，在确定可能的原因特发性RPL. Study基金/竞争兴趣（S）：这项研究是由科学和技术部科学和工程研究委员会（EMR/2014/000145）和国家研究所生殖健康内部基金（RA/872/01-2020）。所有作者均声明没有利益冲突。
STUDY QUESTION: What is the sperm DNA methylation status of imprinted genes in male partners from couples experiencing recurrent pregnancy loss (RPL)?SUMMARY ANSWER: Aberrations in sperm DNA methylation status of several imprinted genes, such as insulin like growth factor-2-H19 differentially methylated region (IGF2-H19 DMR), intergenic differentially methylated region (IG-DMR), mesoderm specific transcript (MEST), zinc finger protein which regulates apoptosis and cell cycle arrest (ZAC), DMR in intron 10 of KCNQ1 gene (KvDMR), paternally expressed gene 3 (PEG3) and paternally expressed gene 10 (PEG10), as well as decreased sperm global 5-methylcytosine (5mC) levels, are associated with RPL.WHAT IS KNOWN ALREADY: RPL is defined as loss of two or more pregnancies, affecting 1-2% of couples of reproductive age. Although there are several maternal and paternal aetiological factors contributing to RPL, nearly 50% of the cases remain idiopathic. Thus, there is a need to identify putative paternal factors that could be contributing towards pregnancy loss in cases of idiopathic RPL.STUDY DESIGN, SIZE, DURATION: In this case-control study, 112 couples undergoing RPL with no identifiable cause were recruited from September 2015 to May 2018. The control group comprised of 106 healthy proven fertile couples with no history of infertility or miscarriage.PARTICIPANTS/MATERIALS, SETTING, METHODS: In this study, we investigated the paternal genetic and epigenetic factors that could be associated with RPL. We studied DNA methylation, by pyrosequencing, of selected imprinted genes implicated in embryo development, such as IGF2-H19 DMR, IG-DMR, MEST, ZAC, KvDMR, PEG3, PEG10 and small nuclear ribonucleoprotein polypeptide N (SNRPN) in sperm of men whose partners present RPL. Global DNA methylation in sperm was evaluated by studying 5mC content and long interspersed nuclear element I (LINEI) promoter methylation. We also studied polymorphisms by pyrosequencing in the IGF2 HI9 DMR as well in the IGF2 promoter in both groups.MAIN RESULTS AND THE ROLE OF CHANCE: In the RPL group, we found a significant decrease in the global sperm 5mC levels and significant decrease in DNA methylation at three CpG sites in LINE! promoter. For IGF2 HI 9 DMR and IG DMR, a significant decrease in sperm DNA methylation at specific CpG sites was observed in RPL group. For maternally imprinted genes like MEST, ZAC, KvDMR, PEG3 and PEG10 hypermethylation was noted. Polymorphism studies for IGF2 H19 DMR and IGF2 revealed significant differences in the genotypic frequencies in males.IMITATIONS, REASONS FOR CAUTION: In this study, we analysed the methylation levels of selected candidate imprinted genes implicated in embryo development. Detection of methylation changes occurring at the genome-wide level may reveal further candidate genes having a better distinction between the control and study groups.WIDER IMPLICATIONS OF THE FINDINGS: Our study demonstrates that certain polymorphisms and aberrant sperm methylation status in imprinted genes are associated with RPL and could contribute to the aetiology of RPL. This study suggests that investigation of paternal genetic and epigenetic factors could be useful in identification of possible causes of idiopathic RPL.STUDY FUNDING/COMPETING INTEREST(S): This study was funded by Department of Science and Technology-Science and Engineering Research Board (EMR/2014/000145) and National Institute for Research in Reproductive Health intramural funds (RA/872/01-2020). All authors declare no conflict of interest.