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Abstract：
BACKGROUND: Recurrent spontaneous abortion (RSA) is a challenging condition that affects the health of women both physically and mentally, but its pathogenesis and treatment have yet to be studied in detail. In recent years, Wharton\'s jelly-derived mesenchymal stem cells (WJ-MSCs) have been shown to be effective in treating various diseases. Current understanding of RSA treatment using WJ-MSCs is limited, and the exact mechanisms of WJ-MSCs action in RSA remains largely unclear. In this study, we explored the decidual deficiencies in RSA and the therapeutic potential of WJ-MSCs at single-cell resolution.
METHODS: Three mouse models were established: a normal pregnancy group, an RSA group, and a WJ-MSC treatment group. Decidual tissue samples were collected for single-cell RNA sequencing (scRNA-seq) and functional verification, including single-cell resolution in situ hybridization on tissues (SCRINSHOT) and immunofluorescence.
RESULTS: We generated a single-cell atlas of decidual tissues from normal pregnant, RSA, and WJ-MSC-treated mice and identified 14 cell clusters in the decidua on day 14. Among these cell populations, stromal cells were the most abundant cell clusters in the decidua, and we further identified three novel subclusters (Str_0, Str_1, and Str_2). We also demonstrated that the IL17 and TNF signaling pathways were enriched for upregulated DEGs of stromal cells in RSA mice. Intriguingly, cell-cell communication analysis revealed that Str_1 cell-related gene expression was greatly reduced in the RSA group and rescued in the WJ-MSC treatment group. Notably, the interaction between NK cells and other cells in the RSA group was attenuated, and the expression of Spp1 (identified as an endometrial toleration-related marker) was significantly reduced in the NK cells of the RSA group but could be restored by WJ-MSC treatment.
CONCLUSIONS: Herein, we implemented scRNA-seq to systematically evaluate the cellular heterogeneity and transcriptional regulatory networks associated with RSA and its treatment with WJ-MSCs. These data revealed potential therapeutic targets of WJ-MSCs to remodel the decidual subpopulations in RSA and provided new insights into decidua-derived developmental defects at the maternal-foetal interface.
METHODS: Three mouse models were established: a normal pregnancy group, an RSA group, and a WJ-MSC treatment group. Decidual tissue samples were collected for single-cell RNA sequencing (scRNA-seq) and functional verification, including single-cell resolution in situ hybridization on tissues (SCRINSHOT) and immunofluorescence.
RESULTS: We generated a single-cell atlas of decidual tissues from normal pregnant, RSA, and WJ-MSC-treated mice and identified 14 cell clusters in the decidua on day 14. Among these cell populations, stromal cells were the most abundant cell clusters in the decidua, and we further identified three novel subclusters (Str_0, Str_1, and Str_2). We also demonstrated that the IL17 and TNF signaling pathways were enriched for upregulated DEGs of stromal cells in RSA mice. Intriguingly, cell-cell communication analysis revealed that Str_1 cell-related gene expression was greatly reduced in the RSA group and rescued in the WJ-MSC treatment group. Notably, the interaction between NK cells and other cells in the RSA group was attenuated, and the expression of Spp1 (identified as an endometrial toleration-related marker) was significantly reduced in the NK cells of the RSA group but could be restored by WJ-MSC treatment.
CONCLUSIONS: Herein, we implemented scRNA-seq to systematically evaluate the cellular heterogeneity and transcriptional regulatory networks associated with RSA and its treatment with WJ-MSCs. These data revealed potential therapeutic targets of WJ-MSCs to remodel the decidual subpopulations in RSA and provided new insights into decidua-derived developmental defects at the maternal-foetal interface.
摘要：
背景：复发性自然流产（RSA）是一种具有挑战性的疾病，会影响女性的身心健康，但其发病机制和治疗方法还有待详细研究。近年来,沃顿胶质来源的间充质干细胞（WJ-MSCs）已被证明可有效治疗各种疾病。目前对使用WJ-MSCs进行RSA治疗的了解有限，WJ-MSCs在RSA中的作用机制尚不清楚。在这项研究中,我们探索了RSA的蜕膜缺陷和WJ-MSCs在单细胞分辨率下的治疗潜力。
方法：建立三种小鼠模型：正常妊娠组，RSA组,和WJ-MSC治疗组。收集蜕膜组织样本进行单细胞RNA测序（scRNA-seq）和功能验证，包括组织上的单细胞分辨率原位杂交（SCRINSHOT）和免疫荧光。
结果：我们生成了一个正常孕妇蜕膜组织的单细胞图谱，RSA,和WJ-MSC处理的小鼠,并在第14天鉴定蜕膜中的14个细胞簇。在这些细胞群中，基质细胞是蜕膜中最丰富的细胞簇，我们进一步确定了三个新的子簇(Str_0、Str_1和Str_2)。我们还证明了IL17和TNF信号通路富集了RSA小鼠基质细胞的上调的DEGs。有趣的是，细胞通讯分析显示，Str_1细胞相关基因表达在RSA组中大大降低，而在WJ-MSC治疗组中被挽救。值得注意的是,RSA组中NK细胞与其他细胞之间的相互作用减弱，在RSA组的NK细胞中，Spp1（被鉴定为子宫内膜耐受相关标志物）的表达显着降低，但可以通过WJ-MSC治疗恢复。
结论：此处，我们使用scRNA-seq系统评估了与RSA相关的细胞异质性和转录调控网络,并使用WJ-MSCs治疗.这些数据揭示了WJ-MSCs的潜在治疗靶标，以重塑RSA中的蜕膜亚群，并为母婴界面处的蜕膜衍生发育缺陷提供了新的见解。
方法：建立三种小鼠模型：正常妊娠组，RSA组,和WJ-MSC治疗组。收集蜕膜组织样本进行单细胞RNA测序（scRNA-seq）和功能验证，包括组织上的单细胞分辨率原位杂交（SCRINSHOT）和免疫荧光。
结果：我们生成了一个正常孕妇蜕膜组织的单细胞图谱，RSA,和WJ-MSC处理的小鼠,并在第14天鉴定蜕膜中的14个细胞簇。在这些细胞群中，基质细胞是蜕膜中最丰富的细胞簇，我们进一步确定了三个新的子簇(Str_0、Str_1和Str_2)。我们还证明了IL17和TNF信号通路富集了RSA小鼠基质细胞的上调的DEGs。有趣的是，细胞通讯分析显示，Str_1细胞相关基因表达在RSA组中大大降低，而在WJ-MSC治疗组中被挽救。值得注意的是,RSA组中NK细胞与其他细胞之间的相互作用减弱，在RSA组的NK细胞中，Spp1（被鉴定为子宫内膜耐受相关标志物）的表达显着降低，但可以通过WJ-MSC治疗恢复。
结论：此处，我们使用scRNA-seq系统评估了与RSA相关的细胞异质性和转录调控网络,并使用WJ-MSCs治疗.这些数据揭示了WJ-MSCs的潜在治疗靶标，以重塑RSA中的蜕膜亚群，并为母婴界面处的蜕膜衍生发育缺陷提供了新的见解。
