CS Mott Center/Ob/Gyn Department, Wayne State University (WSU), Detroit, Michigan, USA.
Reproductive Stress Inc, Grosse Pointe Farms, Michigan, USA.
Birth Defects Res. 2024 Nov;116(11):e2409. doi: 10.1002/bdr2.2409.
Miscarriages cause a greater loss-of-life than cardiovascular diseases, but knowledge about environmentally induced miscarriages is limited. Cultured naïve pluripotent embryonic stem cells (ESC) differentiate into extra-embryonic endoderm/extraembryonic endoderm (XEN) or formative pluripotent ESC, during the period emulating maximal miscarriage of peri-implantation development. In previous reports using small marker sets, hyperosmotic sorbitol, or retinoic acid (RA) decreased naïve pluripotency and increased XEN by FACS quantitation.
Bulk and single cell (sc)RNAseq analyses of two cultured ESC lines was done, corroborated by qPCR. Transcriptomic responses were analyzed of cultured ESC stressed by Sorbitol, with Leukemia inhibitory factor (LIF + ; stemness growth factor), RA without LIF to control for XEN induction, and compared with normal differentiation (LIF - , ND).
Sorbitol and RA increase subpopulations of 2-cell embryo-like (2CEL) and XEN sub-lineages; primitive, parietal, and visceral endoderm (VE) cells and suppress formative pluripotency, imbalancing alternate lineage choices of initial naïve pluripotent cultured ESC compared with ND. Although bulk RNAseq and gene ontology (GO) group analyses suggest that stress induces anterior VE-head organizer and placental markers, scRNAseq reveals relatively few cells. But VE and placental markers/cells were in adjacent stressed cell clusters in the UMAP, like recent, normal UMAP of conceptuses. UMAPs show that dose-dependent stress overrides stemness to force premature lineage imbalance.
Hyperosmotic stress, and other toxicological stresses, like drugs with active ingredient RA, may cause premature, lineage imbalance, resulting in miscarriages or birth defects.
流产导致的死亡人数超过心血管疾病，但人们对环境引起的流产知之甚少。在模拟着床前发育过程中最大流产的时期，未成熟的多能胚胎干细胞（ESC）在体外培养中分化为胚外内胚层/胚外内胚层（XEN）或形成多能 ESC。在以前使用小标记集、高渗山梨醇或维甲酸（RA）的报告中，通过 FACS 定量减少了未成熟的多能性并增加了 XEN。
对两种培养的 ESC 系进行了批量和单细胞（sc）RNAseq 分析，并通过 qPCR 进行了验证。分析了山梨醇培养的 ESC 的转录组反应，同时使用白血病抑制因子（LIF+；干细胞生长因子）处理，而 RA 没有 LIF 来控制 XEN 诱导，并与正常分化（LIF-，ND）进行比较。
山梨醇和 RA 增加了 2 细胞胚胎样（2CEL）和 XEN 亚系的亚群；原始、壁内和内脏内胚层（VE）细胞，并抑制了形成多能性，与 ND 相比，初始未成熟培养的 ESC 选择替代谱系的能力失衡。尽管批量 RNAseq 和基因本体论（GO）组分析表明应激诱导了前 VE-头组织者和胎盘标记物，但 scRNAseq 揭示了相对较少的细胞。但是，VE 和胎盘标记物/细胞在 UMAP 中与相邻的应激细胞簇相邻，类似于最近的正常概念 UMAP。UMAP 显示，剂量依赖性应激会破坏干细胞特性，迫使早期谱系失衡。
高渗应激和其他毒性应激，如含有活性成分 RA 的药物，可能导致过早的、谱系失衡，导致流产或出生缺陷。
