Abstract：
BACKGROUND: Factors contributing to recurrent pregnancy loss (RPL) in more than half of the cases are still unknown. The incidence and societal impact of this condition requires urgent elucidation of the mechanisms behind it, which could aid in significant improvement of clinical management.
METHODS: Using a highly efficient in-solution digestion method and label-free data-independent LC-MS/MS acquisition with ion mobility, we performed comparative proteomics analysis of the decidua tissues from 19 RPL patients and 10 controls. Differentially abundant proteins (DAPs) were compared and correlated with 3 publicly available transcriptomic datasets and the expression of selected biomarkers was tested by qPCR in decidua and chorionic villi from an extended cohort.
RESULTS: From 1952 proteins identified based on ≥2 peptides, the statistically significant difference in abundance (Anova p ≤ 0.05) and fold change ≥1.2 showed 85 proteins. Pathway analysis using Reactome, KEGG and Wiki pathways identified enrichment of \"Signaling by ROBO receptors\", \"RNA degradation\" and \"Cytoplasmic Ribosomal Proteins\". The correlation between protein and gene expression in decidua revealed that the down-regulated ribosomal proteins in our dataset (RPS15, RPS17, RPL27A, RPL35A and RPL18) showed the same regulation trend at the mRNA level, which was later confirmed for transcripts of RPS15 and RPL18 in our cohort.
CONCLUSIONS: Our data suggests that the potential causes of RPL from the maternal side could be associated with impaired RNA processing machinery. Furthermore, the list of DAPs in RPL opens future investigations in terms of screening novel gene variants predisposing to pregnancy failure and developing biomarkers for RPL risk.
METHODS: Using a highly efficient in-solution digestion method and label-free data-independent LC-MS/MS acquisition with ion mobility, we performed comparative proteomics analysis of the decidua tissues from 19 RPL patients and 10 controls. Differentially abundant proteins (DAPs) were compared and correlated with 3 publicly available transcriptomic datasets and the expression of selected biomarkers was tested by qPCR in decidua and chorionic villi from an extended cohort.
RESULTS: From 1952 proteins identified based on ≥2 peptides, the statistically significant difference in abundance (Anova p ≤ 0.05) and fold change ≥1.2 showed 85 proteins. Pathway analysis using Reactome, KEGG and Wiki pathways identified enrichment of \"Signaling by ROBO receptors\", \"RNA degradation\" and \"Cytoplasmic Ribosomal Proteins\". The correlation between protein and gene expression in decidua revealed that the down-regulated ribosomal proteins in our dataset (RPS15, RPS17, RPL27A, RPL35A and RPL18) showed the same regulation trend at the mRNA level, which was later confirmed for transcripts of RPS15 and RPL18 in our cohort.
CONCLUSIONS: Our data suggests that the potential causes of RPL from the maternal side could be associated with impaired RNA processing machinery. Furthermore, the list of DAPs in RPL opens future investigations in terms of screening novel gene variants predisposing to pregnancy failure and developing biomarkers for RPL risk.
摘要：
背景：在超过一半的病例中导致复发性妊娠丢失（RPL）的因素仍然未知。这种情况的发生率和社会影响需要紧急阐明其背后的机制，这可以帮助显著改善临床管理。
方法：使用高效的溶液中消化方法和具有离子迁移率的无标记数据无关LC-MS/MS采集，我们对19例RPL患者和10例对照的蜕膜组织进行了比较蛋白质组学分析.将差异丰度蛋白（DAP）与3个公开可用的转录组数据集进行比较并相关，并通过qPCR在来自扩展队列的蜕膜和绒毛膜绒毛中测试了所选生物标志物的表达。
结果：从1952年基于≥2肽鉴定的蛋白质，丰度（Anovap≤0.05）和倍数变化≥1.2的统计学差异显示有85种蛋白质。使用反应组的路径分析，KEGG和Wiki通路确定了“ROBO受体信号传导”的富集，“RNA降解”和“细胞质核糖体蛋白质”。蜕膜中蛋白质和基因表达之间的相关性表明，在我们的数据集中，核糖体蛋白下调（RPS15，RPS17，RPL27A，RPL35A和RPL18)在mRNA水平上表现出相同的调控趋势,后来在我们的队列中证实了RPS15和RPL18的转录本。
结论：我们的数据表明，母体RPL的潜在原因可能与RNA加工机制受损有关。此外，RPL中的DAP列表为筛选易导致妊娠失败的新基因变异和开发RPL风险的生物标志物开辟了未来的研究.
方法：使用高效的溶液中消化方法和具有离子迁移率的无标记数据无关LC-MS/MS采集，我们对19例RPL患者和10例对照的蜕膜组织进行了比较蛋白质组学分析.将差异丰度蛋白（DAP）与3个公开可用的转录组数据集进行比较并相关，并通过qPCR在来自扩展队列的蜕膜和绒毛膜绒毛中测试了所选生物标志物的表达。
结果：从1952年基于≥2肽鉴定的蛋白质，丰度（Anovap≤0.05）和倍数变化≥1.2的统计学差异显示有85种蛋白质。使用反应组的路径分析，KEGG和Wiki通路确定了“ROBO受体信号传导”的富集，“RNA降解”和“细胞质核糖体蛋白质”。蜕膜中蛋白质和基因表达之间的相关性表明，在我们的数据集中，核糖体蛋白下调（RPS15，RPS17，RPL27A，RPL35A和RPL18)在mRNA水平上表现出相同的调控趋势,后来在我们的队列中证实了RPS15和RPL18的转录本。
结论：我们的数据表明，母体RPL的潜在原因可能与RNA加工机制受损有关。此外，RPL中的DAP列表为筛选易导致妊娠失败的新基因变异和开发RPL风险的生物标志物开辟了未来的研究.
