Advanced paternal age (APA) is associated with adverse outcomes to offspring health, including increased risk for neurodevelopmental disorders. The aim of this study was to investigate the methylome and transcriptome of the first two early embryonic tissue lineages, the inner cell mass (ICM) and the trophectoderm (TE), from human blastocysts in association with paternal age and disease risk. High quality human blastocysts were donated with patient consent from donor oocyte IVF cycles from either APA (≥ 50 years) or young fathers. Blastocysts were mechanically separated into ICM and TE lineage samples for both methylome and transcriptome analyses. Significant differential methylation and transcription was observed concurrently in ICM and TE lineages of APA-derived blastocysts compared to those from young fathers. The methylome revealed significant enrichment for neuronal signaling pathways, as well as an association with neurodevelopmental disorders and imprinted genes, largely overlapping within both the ICM and TE lineages. Significant enrichment of neurodevelopmental signaling pathways was also observed for differentially expressed genes, but only in the ICM. In stark contrast, no significant signaling pathways or gene ontology terms were identified in the trophectoderm. Despite normal semen parameters in aged fathers, these significant molecular alterations can adversely contribute to downstream impacts on offspring health, in particular neurodevelopmental disorders like autism spectrum disorder and schizophrenia. An increased risk for neurodevelopmental disorders is well described in children conceived by aged fathers. Using blastocysts derived from donor oocyte IVF cycles to strategically control for maternal age, our data reveals evidence of methylation dysregulation in both tissue lineages, as well as transcription dysregulation in neurodevelopmental signaling pathways associated with APA fathers. This data also reveals that embryos derived from APA fathers do not appear to be compromised for initial implantation potential with no significant pathway signaling disruption in trophectoderm transcription. Collectively, our work provides insights into the complex molecular mechanisms that occur upon paternal aging during the first lineage differentiation in the preimplantation embryo. Early expression and epigenetic markers of APA-derived preimplantation embryos highlight the susceptibility of the future fetus to adverse health outcomes.

中文翻译：

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父亲衰老早在第一个胚胎组织谱系分化时就影响表达和表观遗传标记

高龄父亲（APA）与后代健康的不良后果相关，包括增加神经发育障碍的风险。本研究的目的是调查人类囊胚中前两个早期胚胎组织谱系（内细胞团（ICM）和滋养外胚层（TE））的甲基化组和转录组与父亲年龄和疾病风险的关系。高质量的人类囊胚是在患者同意的情况下从 APA（≥ 50 岁）或年轻父亲的供卵母细胞 IVF 周期中捐献的。将囊胚机械分离成 ICM 和 TE 谱系样本，用于甲基化组和转录组分析。与来自年轻父亲的囊胚相比，在 APA 衍生的囊胚的 ICM 和 TE 谱系中同时观察到显着差异的甲基化和转录。甲基化组揭示了神经元信号通路的显着富集，以及与神经发育障碍和印记基因的关联，这些基因在 ICM 和 TE 谱系中很大程度上重叠。还观察到差异表达基因的神经发育信号通路显着富集，但仅限于 ICM。形成鲜明对比的是，在滋养外胚层中没有发现显着的信号通路或基因本体术语。尽管年老父亲的精液参数正常，但这些显着的分子改变可能会对后代健康产生不利影响，特别是自闭症谱系障碍和精神分裂症等神经发育障碍。高龄父亲所生的孩子患神经发育障碍的风险增加，这一点已有充分的描述。使用来自供体卵母细胞 IVF 周期的囊胚来战略性地控制母亲年龄，我们的数据揭示了两个组织谱系中甲基化失调的证据，以及与 APA 父亲相关的神经发育信号通路中的转录失调的证据。该数据还表明，源自 APA 父亲的胚胎似乎并未因初始植入潜力而受到损害，滋养外胚层转录中没有显着的途径信号中断。总的来说，我们的工作提供了对植入前胚胎第一次谱系分化过程中父系衰老时发生的复杂分子机制的见解。 APA 衍生的植入前胚胎的早期表达和表观遗传标记突显了未来胎儿对不良健康结果的易感性。