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scRNA-seq of preeclamptic trophoblasts identifies EBI3, COL17A1, miR-27a-5p, and miR-193b-5p as hypoxia markers: validation of neuradapt as a superior mimetic to cobalt chloride
Background. Preeclampsia (PE) complicates 2–8% of pregnancies and involves placental hypoxia and HIF-pathway activation, especially in early-onset PE (eoPE). Chemical mimetics like cobalt(II) chloride (CoCl2) and oxyquinoline derivatives model trophoblast hypoxia in vitro, yet their fidelity in recapitulating PE gene profiles remains unclear. Integrating patient tissue analyses with experimental models may reveal common markers and validate physiologically relevant paradigms.
Methods. We analyzed scRNA-seq data from 10 eoPE, 7 late-onset PE, and matched control placentas, identifying villous cytotrophoblast, syncytiotrophoblast, and extravillous trophoblast (EVT). BeWo b30 cells were treated for 24 h with CoCl2 (300 µM) or the oxyquinoline derivative neuradapt (5 µM) to induce hypoxia. RNA-seq with qPCR validation and small RNA-seq quantified mRNA and microRNA changes; PROGENy inferred pathway activities.
Results. scRNA-seq revealed highest hypoxia activation in eoPE, with EVT showing maximum activity. Nine genes were upregulated across all trophoblast types (EBI3, CST6, FN1, RFK, COL17A1, LDHA, PKP2, RPS4Y1, RPS26). In vitro, neuradapt induced more specific hypoxia responses than CoCl2 (1,284 vs. 3,032 differentially expressed genes). Critically, EBI3, FN1, and COL17A1 showed concordant upregulation in tissue and neuradapt-treated cells, whereas CoCl2 produced opposite patterns. MicroRNAs hsa-miR-27a-5p and hsa-miR-193b-5p were consistently elevated in both models; 3'-isoforms of hsa-miR-9-5p and hsa-miR-92b-3p were identified as hypoxia-associated.
Conclusions. EBI3, COL17A1, miR-27a-5p, and miR-193b-5p emerge as trophoblast hypoxia markers. Neuradapt (a selective HIF-prolyl hydroxylase inhibitor) provides a more physiologically relevant in vitro model than CoCl2, recapitulating transcriptomic signatures observed in PE placentas. This integrated approach advances understanding of PE pathophysiology and therapeutic targeting.