背景:精子发生是一个高度调节且复杂的过程,其中DNA甲基化起着至关重要的作用。本研究旨在探讨弱精子症(AS)患者和健康对照(HCs)患者精子DNA的差异甲基化谱,那些患有少弱精子症(OAS)和HCs的人,以及AS患者和OAS患者。
结果:收集了5例AS患者的精液样本和临床数据,5名OAS患者,和六个年龄匹配的HC。进行减少代表性的亚硫酸氢盐测序(RRBS)以鉴定不同类型的患者和HC中的精细胞中的差异甲基化区域(DMRs)。在AS和HC之间共检测到6520、28,019和16,432个DMR,OAS和HC,以及AS和OAS集团,分别。这些DMR主要位于基因体内,并定位到相应组中的2868、9296和9090个基因。值得注意的是,每组12、9和8个DMRs与精子发生和男性不育密切相关。此外,BDNF,SMARCB1,PIK3CA,和DDX27;RBMX和SPATA17;ASZ1,CDH1和CHDH被鉴定为每组中的强差异甲基化候选基因,分别。同时,AS中DMR相关基因的GO分析与HC小组揭示了蛋白质结合,细胞质,和转录(DNA模板)是生物过程(BP)中最丰富的术语,细胞成分(CC),和分子功能(MF),分别。同样,在OAS与HC和ASvs.美洲国家组织,GO分析显示蛋白质结合,核,和转录(DNA模板)作为BP中最丰富的术语,CC,MF,分别。最后,对DMR注释基因和这些启动子基因的KEGG分析表明,在所有三组中,代谢途径的相关性最为显著.
结论:目前的研究结果揭示了AS患者精子DNA甲基化模式与HC和OASvs.HC组,特别是在AS患者和OAS患者之间。除了差异富集的代谢途径外,鉴定与精子发生和男性不育相关的关键基因可能有助于揭示不同类型异常精子参数的潜在发病机理。
BACKGROUND: Spermatogenesis is a highly regulated and complex process in which DNA methylation plays a crucial role. This study aimed to explore the differential methylation profiles in sperm DNA between patients with asthenospermia (AS) and healthy controls (HCs), those with oligoasthenospermia (OAS) and HCs, and patients with AS and those with OAS.
RESULTS: Semen samples and clinical data were collected from five patients with AS, five patients with OAS, and six age-matched HCs. Reduced representation bisulfite sequencing (RRBS) was performed to identify differentially methylated regions (DMRs) in sperm cells among the different types of patients and HCs. A total of 6520, 28,019, and 16,432 DMRs were detected between AS and HC, OAS and HC, and AS and OAS groups, respectively. These DMRs were predominantly located within gene bodies and mapped to 2868, 9296, and 9090 genes in the respective groups. Of note, 12, 9, and 8 DMRs in each group were closely associated with spermatogenesis and male infertility. Furthermore, BDNF, SMARCB1, PIK3CA, and DDX27; RBMX and SPATA17; ASZ1, CDH1, and CHDH were identified as strong differentially methylated candidate genes in each group, respectively. Meanwhile, the GO analysis of DMR-associated genes in the AS vs. HC groups revealed that protein binding, cytoplasm, and transcription (DNA-templated) were the most enriched terms in the biological process (BP), cellular component (CC), and molecular function (MF), respectively. Likewise, in both the OAS vs. HC and AS vs. OAS groups, GO analysis revealed protein binding, nucleus, and transcription (DNA-templated) as the most enriched terms in BP, CC, and MF, respectively. Finally, the KEGG analysis of DMR-annotated genes and these genes at promoters suggested that metabolic pathways were the most significantly associated across all three groups.
CONCLUSIONS: The current study results revealed distinctive sperm DNA methylation patterns in the AS vs. HC and OAS vs. HC groups, particularly between patients with AS and those with OAS. The identification of key genes associated with spermatogenesis and male infertility in addition to the differentially enriched metabolic pathways may contribute to uncovering the potential pathogenesis in different types of abnormal sperm parameters.