Single cell RNA-sequencing reveals BHLHE40 and NDRG1 as key regulatory molecules modulating the trophoblastic senescence in preeclampsia

INTRODUCTION: Preeclampsia (PE) is a severe obstetric complication characterized by trophoblastic dysfunction and compromised placental development. Cellular senescence has been notably reported in PE trophoblasts, which contributes to placental dysfunction. However, the underlying molecular mechanisms and key regulatory targets governing this process remain poorly understood. METHODS: We analyzed PE-associated single-cell RNA sequencing (GSE173193), ChIP detection (GSE24129), and bulk transcriptome (GSE75010) datasets from GEO database. Through integrated analysis of trophoblast-specific differential expression, weighted gene co-expression network analysis (WGCNA), and senescence-related gene profiles, we identified BHLHE40 and NDRG1 as key regulatory molecules. Their clinical predictive value was comprehensively assessed via the nomogram model. Bioinformatic findings were systematically validated in human PE placentas and LPS-induced PE rat model. The functional effects of BHLHE40 and NDRG1 overexpression on trophoblastic senescence were extensively evaluated in vitro. RESULTS: BHLHE40 and NDRG1 emerged as crucial regulators of trophoblastic senescence in PE, demonstrating significant predictive value for adverse pregnancy outcomes. Both molecules showed consistent aberrant overexpression in human PE placentas and LPS-induced PE rat placentas. Importantly, their overexpression significantly accelerated cellular senescence in trophoblasts in vitro. DISCUSSION: Through integrated bioinformatic analysis and experimental validation, we reveal BHLHE40 and NDRG1 as key regulators of abnormal trophoblast senescence in PE, providing new insights into placental dysfunction mechanisms and potential therapeutic targets in PE management.