Endocrine Abstracts

Women who are obese are more likely to suffer early pregnancy loss. Once factor that can contribute to this is endometrial dysfunction however, we are limited in our understanding of how stressors or treatments, may alter endometrial function. Mimicking the dynamic in vivo exposure of the endometrium to these stressors is difficult using in vitro static culture systems. The aim of this study was to use a microfluidic approach to mimic exposure of the endometrial epithelium to factors in maternal circulation representative of different metabolic states to identify pathways that may contribute to endometrial dysfunction. Ishikawa cells were seeded (1,000,000/mL) in microfluidic devices (n = 3) and exposed to one of the following treatments at a rate of 1 ul/mL for 24 hours: 1) Control, 2) Vehicle Control, 3) Leptin (L-non-obese; 11 ng/ml), 4) Leptin (LO-obese; 35 ng/ml), 5) Leptin obese + Metformin (LOM-500 ug/ml), or 6) Leptin (LOMA-obese 35ng/ml) + Metformin (500 ug/ml) + Adiponectin (2.5 ug/ml). Following RNA sequencing of extracted cells, differentially expressed genes (DEGs) compared to VC were identified and subjected to overrepresentation analysis using Webgestalt. In total 544 DEGs were identified in cells treated with normal physiological concentrations of leptin and are involved in pathways of protein folding and calcium pathway. In contrast, only 337 were different in LO and included pathways involved in peptide hormone metabolism, biosynthesis, and GABA receptor activation. Only 71 DEGs were in common between these two comparisons. 375 DEGs were different in LOM treated cells. Overrepresented pathways included defective GALNT family members. In LOMA treated cells 381 DEGs were identified with an overrepresentation of transcripts associated with Mucins and O-linked glycosylation. Minimal overlap between all three comparisons (LO, LOM, LOMA) was observed demonstrating treatment-specific changes. Collectively these data have identified pathways to investigate for functional effects on implantation.