C.S. Mott Center for Human Growth and Development, Department of Obstetrics and Gynecology, School of Medicine, Wayne State University, Detroit, MI, 48201, USA.
Department of Biostatistics and Epidemiology, School of Public Health and Health Sciences, University of Massachusetts Amherst, 715 North Pleasant Street, Amherst, MA, USA.
Environ Pollut. 2023 Jul 15;329:121529. doi: 10.1016/j.envpol.2023.121529. Epub 2023 Mar 30.
Non-coding RNA (ncRNA) cargo of extracellular vesicles (EVs) in the male reproductive tract play critical roles in semen quality and emerging evidence suggests their susceptibility to environmental factors. Male phthalate exposures have been linked to poor semen quality, sperm DNA methylation profiles and embryo development; however, there is limited evidence on their potential impact on EV ncRNAs profiles. We evaluated the association between urinary phthalate metabolites and small ncRNAs (sncRNAs) of seminal plasma EVs (spEV) among men receiving clinical infertility care. We conducted sncRNA sequencing of EVs in 96 seminal plasma samples collected from the Sperm Environmental Epigenetics and Development Study (SEEDS). Sequencing reads were mapped to human transcriptome databases using STAR. Urinary metabolite concentrations of thirteen phthalates and two DiNCH, a phthalate alternative, were measured via tandem mass spectrometry. Associations with normalized counts were assessed using EdgeR (FDR<0.05) adjusting for urinary dilution via specific gravity, age, BMI, batch, and biotype-specific total counts. Select metabolites, MEOHP, MECPP, ∑DEHP, MCPP, MCNP, MCOP, were negatively (p < 0.05) correlated with miRNA relative abundance. Similarly, nine metabolites including MEOHP, MECPP, MEHP, MCPP, MHBP, MHiNCH, MiBP, MEHHP, MCOP and ∑DEHP were associated (q < 0.05) with normalized counts from 23 unique ncRNA transcripts (7 miRNAs (pre & mature); 6 tRFs; and 10 piRNAs), most (78%) of which displayed increased expression patterns. miRNA and tRFs gene targets were enriched in vesicle-mediated transport and developmental-related ontology terms, such as tyrosine kinase, head development, and cell morphogenesis. Six genes (MAPK1, BMPR1A/2, PTEN, TGFBR2, TP53 and APP) were present in all the ontology terms and predicted to form protein association networks. piRNAs were annotated to pseudogenes of genes important in EV cargo transfer and embryonic development. This is the first study to associate phthalate exposures to altered spEV sncRNA profiles. Future studies are needed to determine their impact on reproductive outcomes.
外泌体（EVs）中非编码 RNA（ncRNA）载货运送物在男性生殖中的精液质量中起着关键作用，并且有新的证据表明它们容易受到环境因素的影响。男性邻苯二甲酸酯暴露与精液质量差、精子 DNA 甲基化谱和胚胎发育有关；然而，关于它们对 EV ncRNA 谱的潜在影响的证据有限。我们评估了接受临床不孕治疗的男性尿液中邻苯二甲酸酯代谢物与精液外泌体小 ncRNA（sncRNA）之间的关系。我们对 Sperm Environmental Epigenetics and Development Study (SEEDS) 中收集的 96 个精液样本中的 EVs 进行了 sncRNA 测序。使用 STAR 将测序读数映射到人类转录组数据库。通过串联质谱法测量了 13 种邻苯二甲酸酯和 2 种邻苯二甲酸二异壬酯（DiNCH）的尿代谢产物浓度，DiNCH 是邻苯二甲酸酯的替代品。使用 EdgeR（FDR<0.05）调整尿比重、年龄、BMI、批次和生物型特异性总计数来评估与归一化计数的相关性。MEOHP、MECPP、∑DEHP、MCPP、MCNP、MCOP 等选择的代谢物与 miRNA 的相对丰度呈负相关（p<0.05）。同样，9 种代谢物包括 MEOHP、MECPP、MEHP、MCPP、MHBP、MHiNCH、MiBP、MEHHP、MCOP 和 ∑DEHP 与 23 个独特 ncRNA 转录物的归一化计数相关（q<0.05）（7 个 miRNA（前体和成熟）；6 个 tRFs；和 10 个 piRNAs），其中大多数（78%）表现出增加的表达模式。miRNA 和 tRFs 的基因靶标在囊泡介导的运输和发育相关的本体论术语中富集，如酪氨酸激酶、头部发育和细胞形态发生。六个基因（MAPK1、BMPR1A/2、PTEN、TGFBR2、TP53 和 APP）存在于所有本体论术语中，并预测形成蛋白质关联网络。piRNA 被注释到 EV 货物转运和胚胎发育中重要基因的假基因。这是第一项将邻苯二甲酸酯暴露与改变的 spEV sncRNA 谱相关联的研究。需要进一步的研究来确定它们对生殖结果的影响。
