Singh Aditi, Ruden Ximena Lu, Tang Wen, Awonuga Awoniyi Olumide, Ruden Douglas Mark, Korzeniewski Steven James, Puscheck Elizabeth Ella, Feng Hao, Rappolee Daniel Allen
CS Mott Center/Ob/Gyn Department, Wayne State University (WSU), Detroit, MI, USA.
Center for Molecular Medicine and Genetics, WSU, Detroit, MI, USA.
Sci Rep. 2025 Jul 1;15(1):21291. doi: 10.1038/s41598-025-06628-z.
Embryo development is highly susceptible to environmental stressors, contributing significantly to early miscarriages in 70% of human embryos (Cross et al. in Science 266:1508-1518, 1994, Macklon et al. in Hum Reprod Update 8:333-343, 2002). Yet the underlying mechanisms remain poorly understood. Here, we employ mouse embryonic stem cells (ESC) as a model to identify shared "pan-stress" markers for responses to diverse environmental insults implicated in miscarriage (Puscheck et al. in Birth Defects Res 114:1014-1036, 2022, Puscheck et al., in: Leese HJ, Brison DR (eds) Cell Signaling During Mammalian Early Embryo Development, Springer, New York, 2015 ). ESC were exposed to control stimuli and diverse stressors at previously characterized risk levels for growth, mimicking miscarriage risk exposures, then subjected to transcriptomic analysis via RNA sequencing. Surprisingly, we identify a large, substantial set of significantly-changing genes-termed pan-stress genes that also exhibited concordant changes in directionality compared to initial stemness. These genes show significant differential expression across stress conditions, distinguishing weak and strong stressors. Notably, ninety-four genes display significant expression changes under four weaker stressors and normal differentiation conditions, while a twenty-one/ninety-four gene subset exhibits shared significance with the inclusion of two stronger stimuli. Importantly, all identified pan-stress genes, the set of ninety-four and subset of twenty-one, exhibit 100% concordant, highly nonrandom directional changes from the initial stemness. Transcription and secreted factors that might mediate nonrandom concordant stress response were identified. These findings characterize a robust pan-stress response in ESC, suggesting potential biomarkers for miscarriage prediction and testing of underlying mechanisms.
胚胎发育对环境应激源高度敏感，这在70%的人类胚胎早期流产中起了重要作用（Cross等人，《科学》266:1508 - 1518，1994年；Macklon等人，《人类生殖更新》8:333 - 343，2002年）。然而，其潜在机制仍知之甚少。在这里，我们采用小鼠胚胎干细胞（ESC）作为模型，以确定对与流产相关的多种环境损伤作出反应的共同“泛应激”标志物（Puscheck等人，《出生缺陷研究》114:1014 - 1036，2022年；Puscheck等人，见：Leese HJ、Brison DR（编）《哺乳动物早期胚胎发育中的细胞信号传导》，施普林格出版社，纽约，2015年）。将ESC暴露于对照刺激和具有先前确定的生长风险水平的多种应激源下，模拟流产风险暴露，然后通过RNA测序进行转录组分析。令人惊讶的是，我们鉴定出大量显著变化的基因——称为泛应激基因，与初始干性相比，这些基因在方向性上也表现出一致的变化。这些基因在不同应激条件下显示出显著的差异表达，区分了弱应激源和强应激源。值得注意的是，94个基因在四种较弱应激源和正常分化条件下显示出显著的表达变化，而一个21/94基因子集在加入两种较强刺激时表现出共同的显著性。重要的是，所有鉴定出的泛应激基因，即94个基因的集合和21个基因的子集，与初始干性相比都表现出100%一致、高度非随机的方向性变化。鉴定出了可能介导非随机一致应激反应的转录因子和分泌因子。这些发现描绘了ESC中一种强大的泛应激反应，提示了用于流产预测和潜在机制测试的潜在生物标志物。
