KLF4 调节滋养层功能，并与不明原因的复发性自然流产有关。
KLF4 regulates trophoblast function and associates with unexplained recurrent spontaneous abortion.
机构信息
Reproductive Medicine Center, Renmin Hospital of Wuhan University, Wuhan, China.
Department of Pediatrics, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China.
出版信息
J Transl Med. 2024 Oct 10;22(1):922. doi: 10.1186/s12967-024-05707-5.
BACKGROUND
Recurrent spontaneous abortion (RSA) is defined as two or more consecutive spontaneous abortions before 20 weeks with the same spouse [1]. However, approximately 50% of RSA cases of unknown cause are classified as unexplained recurrent spontaneous abortion (URSA). Potential factors include decreased trophoblast cell migration and invasion, leading to impaired placental implantation and maintenance of the normal maternal-fetal interface. However, the mechanism of this pathogenesis remains unknown. In this study, we investigated the potential role and mechanism of KLF4 in regulating URSA by influencing the invasion and migration ability of trophoblast cells.
METHODS
We firstly identified 817 differentially expressed genes by performing a difference analysis of the dataset GSE121950 [2] related to recurrent abortion, and intersected the top 10 genes obtained respectively by the three algorithms: DMNC, MNC, and EPC using Venn Diagram.To detect the expression levels of core genes, villi samples were obtained from normal pregnant women and patients with URSA. RT-qPCR analysis revealed a significant difference in KLF4 mRNA expression and KLF4 was then analyzed. Trophoblast cell lines HTR8 and JEG3 were used to investigate the effect of KLF4 on trophoblastic function. Wound healing and transwell assays was performed to detect the invasion and migration of trophoblast cells. The expression of epithelial-mesenchymal transition(EMT) molecules were detected by RT-qPCR and western blot. Promoter detection and epigenetic modification were detected by chromatin immunoprecipitation (ChIP) assay. Molecular nuclear localization was detected by immunofluorescence and subcellular fractionation. Miscarried mice model was used to study the effects of KLF4 on URSA induced by reduced trophoblast invasion and migration.
RESULTS
KLF4 is highly expressed in the villi of patients with URSA. KLF4 inhibits the expression level of H3R2ME2a in trophoblast cells by regulating the transcriptional level and nuclear translocation of PRMT6, thereby inhibiting the possible regulatory mechanism of trophoblastic invasion and providing a potential treatment strategy for URSA in vivo.
CONCLUSIONS
The KLF4/PRMT6/H3R2ME2a axis regulates mechanisms associated with unexplained recurrent spontaneous abortion by regulating trophoblast function.
背景
复发性自然流产（RSA）是指同一配偶连续发生两次或两次以上 20 周前的自然流产[1]。然而，约 50%的原因不明复发性自然流产（URSA）病例被归类为不明原因的复发性自然流产。潜在因素包括滋养细胞迁移和侵袭能力下降，导致胎盘植入不良和正常的母胎界面维持受损。然而，这种发病机制的机制尚不清楚。在这项研究中，我们通过影响滋养细胞的侵袭和迁移能力，研究了 KLF4 调节 URSA 的潜在作用和机制。
方法
我们首先通过对数据集 GSE121950[2]进行差异分析，鉴定出与复发性流产相关的 817 个差异表达基因，并使用 Venn 图分别对三个算法（DMNC、MNC 和 EPC）获得的前 10 个基因进行交集。为了检测核心基因的表达水平，从正常妊娠妇女和 URSA 患者中获得绒毛样本。RT-qPCR 分析显示 KLF4 mRNA 表达水平存在显著差异，随后对 KLF4 进行分析。使用滋养层细胞系 HTR8 和 JEG3 研究 KLF4 对滋养层功能的影响。通过划痕愈合和 Transwell 实验检测滋养细胞的侵袭和迁移。通过 RT-qPCR 和 Western blot 检测上皮-间充质转化（EMT）分子的表达。通过染色质免疫沉淀（ChIP）实验检测启动子检测和表观遗传修饰。通过免疫荧光和亚细胞分离检测分子核定位。使用流产小鼠模型研究 KLF4 对因滋养细胞侵袭和迁移减少引起的 URSA 的影响。
结果
KLF4 在 URSA 患者的绒毛中高表达。KLF4 通过调节 PRMT6 的转录水平和核转位，抑制滋养细胞中 H3R2ME2a 的表达水平，从而抑制滋养细胞侵袭的可能调节机制，并为体内 URSA 提供潜在的治疗策略。
结论
KLF4/PRMT6/H3R2ME2a 轴通过调节滋养细胞功能调节与不明原因复发性自然流产相关的机制。
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