腺肌病中异常的 TCF21 上调通过增加 PDE4C 的表达来损害子宫内膜蜕膜化。

Aberrant TCF21 upregulation in adenomyosis impairs endometrial decidualization by increasing PDE4C expression.

机构信息

International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200030, China; Shanghai Key Laboratory of Embryo Original Diseases, Shanghai 200030, China; Municipal Key Clinical Specialty, Shanghai 200030, China.

出版信息

Biochim Biophys Acta Mol Basis Dis. 2025 Jan;1871(1):167526. doi: 10.1016/j.bbadis.2024.167526. Epub 2024 Sep 25.

DOI:10.1016/j.bbadis.2024.167526

PMID:39326465

Abstract

BACKGROUND

Impaired decidualization is a major cause of infertility in patients with adenomyosis (AM). However, the effect of transcription factor 21 (TCF21) on AM and the underlying mechanism of associated-impaired decidualization remain unclear. The aim of this study was to investigate the expression of TCF21 in endometrial tissues of AM patients and the specific mechanisms by which it impairs the decidualization of human endometrial stromal cells (HESCs), with a view to improving the reproductive outcome of AM infertile patients.

METHODS

We compared gene expressions via transcriptomics between the control and AM-associated recurrent implantation failure (RIF) groups. qRT-PCR, western blot, and IHC were performed to confirm the expression and location of TCF21 in the endometrium. Furthermore, we confirmed that high expression of TCF21 impairs decidualization by qRT-PCR, immunofluorescence, and western blot. RNA-seq following overexpression of TCF21 in HESCs was conducted to identify TCF21-related molecular changes during in vitro decidualization. Then we performed ChIP-seq/qPCR and dual-luciferase reporter assay to explore the exact interaction between TCF21 and PDE4C. The related downstream mechanisms were further proved using IHC, qRT-PCR, western blot, and ELISA.

RESULTS

According to the RNA-seq analysis, TCF21 expression was remarkably higher in the endometrium of the AM-related RIF group compared to the control group. We confirmed the same results using samples from patients with AM and controls. TCF21 overexpression in HESCs impaired decidualization through suppression of decidual markers and cytoskeleton alterations. The mechanistic analysis revealed that TCF21 inhibited intracellular cAMP levels by directly increasing PDE4C expression and suppressing FOXO1 expression.

CONCLUSIONS

TCF21 compromises decidualization in patients with AM via the PDE4C/cAMP-FOXO1 axis, which offers valuable insights on the pathology of decidualization-related infertility and indicates a potential treatment to improve endometrial receptivity in AM.

摘要

背景

蜕膜化受损是腺肌病（AM）患者不孕的主要原因。然而，转录因子 21（TCF21）对 AM 的影响及其相关蜕膜化受损的潜在机制尚不清楚。本研究旨在探讨 TCF21 在 AM 患者子宫内膜组织中的表达及其对人子宫内膜基质细胞（HESCs）蜕膜化的具体影响机制，以期改善 AM 不孕患者的生殖结局。

方法

通过转录组学比较对照组和 AM 相关复发性种植失败（RIF）组的基因表达。采用 qRT-PCR、Western blot 和 IHC 验证 TCF21 在子宫内膜中的表达和定位。此外，通过 qRT-PCR、免疫荧光和 Western blot 证实 TCF21 高表达可损害蜕膜化。对 HESCs 中转染 TCF21 后进行 RNA-seq，以鉴定体外蜕膜化过程中 TCF21 相关的分子变化。然后进行 ChIP-seq/qPCR 和双荧光素酶报告基因检测，以探讨 TCF21 与 PDE4C 之间的确切相互作用。进一步通过 IHC、qRT-PCR、Western blot 和 ELISA 验证相关下游机制。

结果

根据 RNA-seq 分析，与对照组相比，AM 相关 RIF 组子宫内膜中 TCF21 的表达明显更高。我们使用 AM 患者和对照组的样本验证了相同的结果。TCF21 在 HESCs 中的过表达通过抑制蜕膜标志物和细胞骨架改变来损害蜕膜化。机制分析表明，TCF21 通过直接增加 PDE4C 的表达和抑制 FOXO1 的表达来抑制细胞内 cAMP 水平。