Abstract：
Recurrent miscarriage (RM) is a distressing pregnancy complication with an unknown etiology. Increasing evidence indicates the relevance of dysregulation of human trophoblast stem cells (hTSCs), which may play a role in the development of RM. However, the potential molecular regulatory mechanism underlying the initiation and maintenance of hTSCs is yet to be fully elucidated. In this study, we performed data analysis and identified Forkhead box M1 (FOXM1) as a potential factor associated with RM. FOXM1 is a typical transcription factor known for its involvement in various pathophysiological processes, while the precise function of FOXM1 functions in hTSCs and RM remains incompletely understood. Utilizing RNA-seq, CUT&Tag, ChIP-qPCR, and sodium bisulfite conversion methods for methylation analysis, we elucidate the underlying regulatory mechanisms of FOXM1 in hTSCs and its implications in RM. Our findings demonstrate the relative high expression of FOXM1 in proliferating cytotrophoblasts (CTBs) compared to differentiated extravillous cytotrophoblasts (EVTs) and syncytiotrophoblasts (STBs). Besides, we provide evidence supporting a significant correlation between FOXM1 downregulation and the incidence of RM. Furthermore, we demonstrate the significant role of FOXM1 in regulating hTSCs proliferation and cell cycle through the transcriptional regulation of CDKN3, CCNB2, CCNA2, MAD2L1 and CDC25C. Notably, we observed a correlation between the downregulation of FOXM1 in RM and hypermethylation in its promoter region. Collectively, these results provide insights into the impact of FOXM1 on trophoblast regulation and offer a novel perspective on RM.
摘要：
复发性流产（RM）是一种病因不明的令人痛苦的妊娠并发症。越来越多的证据表明人滋养干细胞（hTSCs）失调的相关性，这可能在RM的发展中发挥作用。然而,hTSCs启动和维持的潜在分子调控机制尚未完全阐明。在这项研究中,我们进行了数据分析，并确定了叉头盒M1（FOXM1）是与RM相关的潜在因素。FOXM1是一种典型的转录因子，因其参与各种病理生理过程而闻名。而FOXM1功能在hTSCs和RM中的确切功能仍未完全理解。利用RNA-seq，CUT&Tag,ChIP-qPCR,以及用于甲基化分析的亚硫酸氢钠转化方法，我们阐明了FOXM1在hTSCs中的潜在调控机制及其在RM中的意义。我们的发现表明，与分化的绒毛外细胞滋养细胞（EVT）和合胞体滋养细胞（STB）相比，FOXM1在增殖的细胞滋养细胞（CTB）中的相对高表达。此外,我们提供的证据支持FOXM1下调与RM发生率之间存在显著相关性.此外，我们证明了FOXM1通过对CDKN3，CCNB2，CCNA2，MAD2L1和CDC25C的转录调控在调节hTSCs增殖和细胞周期中的重要作用。值得注意的是,我们观察到RM中FOXM1的下调与其启动子区域的高甲基化之间存在相关性。总的来说，这些结果为FOXM1对滋养细胞调节的影响提供了见解，并为RM提供了新的视角。
