Gestational diabetes fetal programming activates the PI3K-Akt and NETosis inflammatory axis in neonatal endothelial colony-forming cells 2259927

Abstract Introduction Gestational diabetes mellitus (GDM) is a critical model for studying the developmental programming of cardiovascular risk (CR), where intrauterine inflammation primes the offspring for future diseases. To understand this early-life predisposition, we investigated the functional phenotype of neonatal ECFCs isolated from cord blood of infants born to women with GDM versus lean controls. Methods We performed a transcriptomic analysis utilizing publicly available bulk RNA sequencing data for 60 individuals (SRA Project PRJNA952510). Quality control and filtering was first applied to the sequencing data, derived from well-characterized neonatal ECFCs (CD31+/vWF+), where all samples met the RNA Integrity Number > 7.0 threshold. Results Our analysis reveals a shift toward a pro-inflammatory and pro-thrombotic phenotype already present in GDM-exposed neonatal ECFCs, indicating the advanced programming of fetal endothelium by the GDM environment. Specifically, the transcriptome displayed robust enrichment for the PI3K-Akt signaling pathway (AKT3, HSP90AA1), acting as a central hub for chronic cellular stress and inflammatory survival. Crucially, pathway analysis demonstrated significant upregulation of genes promoting extracellular matrix remodeling and, subsequently, the highly destructive process of neutrophil extracellular trap (NET) formation (SELE, SPP1). This suggests that neonatal ECFCs are actively upregulating molecules that initiate ECFC-neutrophil crosstalk. Conclusion Our data unveils a novel mechanism where GDM exposure activates an underlying inflammatory nexus (PI3K-Akt) in neonatal ECFCs, directly promoting vascular immunopathology via NETosis at birth. These findings position neonatal ECFCs as dysregulated early drivers of sterile inflammation in GDM-exposed offspring compared to lean-exposed controls. Mechanistically, this effect is mediated by the hyperactivation of the PI3K-Akt/NET axis, identifying this pathway as promising target for early-life intervention to mitigate lifetime CR. Funding Source n/a Topic Categories Computational and Systems Immunology (COMP)