Patient-derived decidual organoids reveal rhG-CSF-associated protective responses involving JAK2/STAT3 signaling in recurrent spontaneous abortion.

Abstract

Background: Recurrent spontaneous abortion (RSA) is a complex reproductive disorder frequently lacks effective therapeutic interventions. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) has shown promise in improving pregnancy outcomes in RSA, but its underlying mechanisms remain unclear. Organoid technology offers new opportunities for disease modeling and mechanistic research in reproductive medicine.

Methods: Decidual tissue samples from RSA patients were utilized to establish decidual organoids using three-dimensional (3D) culture techniques. These organoids were characterized via morphological observation, hematoxylin and eosin (H&E) and Periodic Acid-Schiff (PAS) staining, immunofluorescence, and immunohistochemistry to validate key marker expression and hormone responsiveness. The effects of rhG-CSF on organoid viability were evaluated using luminescence assays. Transcriptomic sequencing and bioinformatics analyses were performed to identify differentially expressed genes and pathways following rhG-CSF stimulation. Protein expression and signaling pathway activation were further assessed via Western blot analysis.

Results: Decidual organoids were successfully established and recapitulated the structural and functional features of primary tissue, including hormone responsiveness and marker expression. rhG-CSF significantly promoted organoid proliferation, enhanced anti-apoptotic and angiogenic capacity, and induced inflammatory responses. Transcriptomic and pathway analyses indicated an enrichment and activation of the JAK2/STAT3 signaling pathway following rhG-CSF treatment. Western blot indicated increased phosphorylation of JAK2 and STAT3, upregulation of PCNA, VEGFA, IL-1β, IL-6, and Bcl-2, and downregulation of Bax in response to rhG-CSF.

Conclusion: A patient-derived decidual organoid model was established and validated to recapitulate in vivo tissue characteristics. In this in vitro RSA model, rhG-CSF appeared to exert protective effects by promoting proliferation, inhibiting apoptosis, and enhancing angiogenesis-responses that are potentially associated with the activation of JAK2/STAT3 signaling. These findings provide new insights and experimental evidence that may inform future RSA treatment strategies.