年龄影响子宫内膜基因表达吗?
使用无监督人工智能方法,我们首次报道了35岁女性子宫内膜基因表达的变化。
女性生育率随年龄增长而下降,主要归因于卵母细胞质量和卵巢储备下降。结合其他证据,一个长期的范例认为年龄不影响子宫内膜功能,并且在子宫内膜研究中没有适当控制年龄.
对来自基因表达综合(GEO)样本库的27名不同年龄女性的子宫内膜转录组数据进行了回顾性分析。结果在来自23-43岁女性的20个子宫内膜样本的独立基因表达数据集中得到验证。
从2016年10月至2019年1月,在GEO进行了系统搜索,以确定涉及不同年龄女性的转录组学研究。包括的样本来自正常排卵,育龄期(23-49岁)女性,月经周期正常,无子宫内膜异位症,在之前的子宫内膜研究中用作对照.我们使用来自这些样本的原始基因表达数据和元数据来研究年龄对子宫内膜基因表达的影响。文件已下载,预处理和探索潜在的混杂变量和异常值。人工智能方法被用来定义年龄组,差异表达和功能分析用于在分子水平上证明和理解年龄对基因表达的影响。在来自23-43岁女性的20个子宫内膜样本的独立基因表达数据集中验证了功能结果。
对最初检索的子宫内膜数据集的分析显示,在大多数研究中,参与者的年龄不可用(33.33%)或可追溯(43.33%)。然而,一项研究适用于年龄分析(GSE4888,n=27,23-49岁).样本根据年龄显示不同的转录组概况,从35年开始。共有5778个差异表达基因和27个显著改变的子宫内膜功能(假发现率(FDR)<0.05)与年龄相关的子宫内膜基因表达变化相关。有趣的是,81.48%的受影响的功能与纤毛过程的上调有关,91个基因参与纤毛运动和纤毛发生。其他功能包括血管内皮生长因子信号通路的失调和由参与细胞周期停滞的37个基因触发的上皮增殖的抑制。血管生成,胰岛素信号和端粒保护。使用非靶向方法在独立的数据集中验证了这些发现;确定了与细胞周期停滞相关的20个上调的纤毛过程(FDR<0.02)和6个下调的功能受年龄的影响。在衰老的其他标志如DNA修复抑制或糖代谢(FDR<0.05)。
本文的基础数据在GEO中可用,ID:GSE4888(主数据集)和GSE102131(验证数据集)。
这项研究规模有限,与大多数子宫内膜转录组学研究一样,全转录组分析考虑了相对较小人群中的近22000个变量。然而,我们的研究包括一个主要样本集和随后的验证集,这些样本集提高了我们结果的可重复性,并为得出年龄影响子宫内膜基因表达的结论提供了合理的证据.需要一项更大的前瞻性研究来控制患者特征,以准确描述与年龄相关的变化,具有较高的样本量和较宽的年龄范围。还需要其他研究来确定子宫内膜老化对不孕症的贡献,以便最终转化为临床环境。
我们的发现支持全基因组功能方法中年龄对子宫内膜的影响,打破人类生殖中的子宫内膜老化模式。据我们所知,这项工作是第一个确定,使用全基因组功能非靶向方法,睫状突是与母亲年龄相关的主要功能失调。这些结果应指导研究社区控制年龄作为子宫内膜基因表达研究中的潜在混杂变量,并在进一步的研究中将子宫内膜老化视为临床上不孕症的潜在原因。报道的功能失调可能导致胚胎植入随着年龄的增长而减少,进一步的研究将证明这种失调是否是某些植入失败的基础。此外,这些功能改变的发现可以进行机械研究,特别是与年龄相关的子宫病变增加。
这项研究由SaludCarlosIII研究所通过MiguelServet计划(CP20/00118)资助,该计划授予由FEDER共同资助的PatriciaDiaz-Gimeno(西班牙政府);以及IVI基金会(1706-FVI-041-PD)。A.D.-P.(FPU/15/01398)和A.P.-L.(FPU18/01777)由科学部的博士前计划研究金授予,创新和大学(西班牙政府)。作者没有任何竞争利益需要声明。
不适用。
Does age affect endometrial gene expression?
Using unsupervised artificial intelligence methods, we report for the first time that endometrial gene expression changes from 35 years of age in women.
Female fertility declines with age, largely attributed to declining oocyte quality and ovarian reserve. Combined with other evidence, a longstanding paradigm holds that age does not affect the endometrial function and age has not been controlled for properly in endometrial studies.
A retrospective in silico analysis was performed of endometrial transcriptomic data from the Gene Expression Omnibus (GEO) sample repository for 27 women of different ages. Results were validated in an independent gene expression dataset of 20 endometrial samples from women aged 23-43 years.
A systematic search was performed in GEO from October 2016 to January 2019 to identify transcriptomic studies involving women of different ages. Included samples were from norm-ovulatory, women of reproductive age (23-49 years) with regular menstrual cycles who were free of endometriosis and used as controls in a previous endometrial study. We used raw gene expression data and metadata from these samples to investigate the effect of age on endometrial gene expression. Files were downloaded, pre-processed and explored for potential confounding variables and outliers. Artificial intelligence methods were applied to define age groups, and differential expression and functional analyses were applied to demonstrate and understand the effect of age on gene expression at the molecular level. Functional results were validated in an independent gene expression dataset of 20 endometrial samples from women aged 23-43 years.
Analysis of the initially retrieved endometrial datasets revealed the age of participants was not available (33.33%) or traceable (43.33%) in most studies. However, one study was suitable for age analysis (GSE4888, n = 27, 23-49 years). Samples showed different transcriptomic profiles according to age, beginning at 35 years. A total of 5778 differentially expressed genes and 27 significantly altered endometrial functions (false discovery rate (FDR) < 0.05) were associated with endometrial gene expression changes related to age. Interestingly, 81.48% of affected functions were related to up-regulation of ciliary processes, with 91 genes involved in cilia motility and ciliogenesis. Other functions included dysregulation of the vascular endothelial growth factor signalling pathway and inhibition of epithelial proliferation triggered by 37 genes involved in cell cycle arrest, angiogenesis, insulin signalling and telomere protection. These findings were validated in an independent dataset using a non-targeted approach; 20 up-regulated ciliary processes (FDR < 0.02) and 6 down-regulated functions related to cell cycle arrest were identified as affected by age, among other hallmarks of ageing such as DNA repair inhibition or sugar metabolism (FDR < 0.05).
Data underlying this article are available in GEO, IDs: GSE4888 (main dataset) and GSE102131 (validation dataset).
This study is limited in size, as are most studies of endometrial transcriptomics where whole-transcriptome analysis considers nearly 22 000 variables in a relatively small population. Yet, our study includes a main sample set and subsequent validation set that enhances reproducibility of our results and provides reasonable evidence for concluding that age affects endometrial gene expression. A larger study prospectively controlling for patient characteristics is needed to accurately describe changes related to age, with a higher sample size and across a wide age range. Additional studies also are necessary to determine the endometrial ageing contribution to infertility for ultimate translation to a clinical setting.
Our findings support an influence of age on the endometrium in a genome-wide functional approach, breaking the endometrial ageing paradigm in human reproduction. To our knowledge, this work is the first to identify, using a genome-wide functional non-targeted approach, ciliary processes as the primary dysregulated function associated with maternal age. These results should guide the research community to control for age as a potential confounding variable in endometrial gene expression studies and to consider endometrial ageing in further studies as a potential cause of infertility in the clinical setting. The reported functional dysregulations could contribute to diminished embryo implantation with age and further studies will demonstrate if such dysregulation underlies some cases of implantation failure. Additionally, the discovery of these functional alterations could enable mechanistic studies, particularly around the age-related increase in uterine pathologies.
This research was funded by the Instituto de Salud Carlos III through Miguel Servet programme (CP20/00118) granted to Patricia Diaz-Gimeno (Spanish Government) co-funded by FEDER; and by IVI Foundation (1706-FIVI-041-PD). A.D.-P. (FPU/15/01398) and A.P.-L. (FPU18/01777) are granted by the pre-doctoral programme fellowship from the Ministry of Science, Innovation and Universities (Spanish Government). The authors do not have any competing interests to declare.
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