多组学分析与实验验证揭示WISP2在复发性流产中的作用
Revealing the Action Mechanism of WISP2 in Miscarriage Based Multi-Omic Analysis and Experimental Validation
摘要
该研究对复发性流产(RM)与正常对照的绒毛组织进行单细胞RNA测序,锁定合体滋养层/绒毛外滋养层(SCT/EVT)中的WISP2,并在20例RM与20例对照的绒毛膜样本中用qRT-PCR、Western blot和免疫组化验证;同时在HTR-8细胞过表达WISP2,开展增殖、迁移与侵袭实验。结果显示RM中WISP2表达异常,体外可促进上皮样表型并伴ERK通路激活,抑制滋养层侵袭与EMT,提示其可能参与RM发病,但仍需体内验证。
原文摘要
Recurrent miscarriage (RM) is a major unresolved issue in reproductive medicine. Although WISP2 is known to participate in multiple cellular processes, its function in trophoblast biology and pregnancy maintenance remains unknown. We performed single-cell RNA sequencing on chorionic villus tissues from RM patients and normal controls to identify cell-type-specific transcriptional alterations, which led to the identification of WISP2 as a candidate gene in the SCT/EVT subpopulation. Fetal membrane samples (chorion) were obtained from Qingdao Women and Children's Hospital's Surgical Outpatient Department: 20 RM patients and 20 healthy controls. Using qRT-PCR, Western blot, and immunohistochemistry to detect gene and protein expression. HTR-8 cells were used to construct WISP2 overexpressing cells, and MTT, wound healing and Transwell experiments were performed for functional experiments. Eleven cell classes in the sample were demonstrated by single-cell analysis. Among these cells were DSCs and SCT/extravillous trophoblasts (EVT). The level of WISP2 is low in SCT/EVT cells from RM patients. In chorionic villus tissues, both WISP2 and E-cadherin were significantly upregulated in the RM group relative to the control group (all p 0.05), suggesting that WISP2 promotes an epithelial-like phenotype accompanied by ERK pathway activation. In summary, WISP2 is dysregulated in RM and suppresses trophoblast invasion and EMT in vitro, suggesting a potential role in RM pathogenesis, though in vivo validation is required.
推荐理由:将单细胞转录组与组织及细胞功能验证结合,聚焦RM中滋养层WISP2,主题核心且提供一定机制线索;但样本与体外证据有限,摘要中表达方向表述不够一致,适合作为RM机制研究的浏览文献。
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