(281) Ovarian Microvascular Endothelial Cell Migration is Dysregulated in PCOS

Arielle S. Yeshua^1,2; Megan A. G. Sage³; Diane M. Duffy³

1. Shady Grove Fertility – Jones Institute, Norfolk, VA, USA

2. Department of Obstetrics and Gynecology, Macon and Joan Brock Virginia Health Sciences at Old Dominion University, Norfolk, VA, USA

3. Department of Biomedical and Translational Sciences, Macon and Joan Brock Virginia Health Sciences at Old Dominion University, Norfolk, VA, USA

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Abstract Text:

Polycystic Ovary Syndrome (PCOS) is a common endocrine disorder affecting 6-12% of women worldwide, characterized by irregular ovulation, polycystic ovarian morphology, and elevated androgens. In addition to infertility, women with PCOS are at a higher risk of cardiovascular disease and pregnancy-related cardiovascular complications, even independent of obesity and insulin resistance. These risks may be linked to endothelial cell dysfunction, which contributes to both macro and microvascular systemic impairments. As angiogenesis is essential for follicular growth and successful ovulation, ovarian endothelial function dysfunction may have significant implications in PCOS. VEGFA and prostaglandin E2 (PGE2) are known to regulate ovarian microvascular endothelial cell function. This study investigates differences in ovarian microvascular endothelial cell responses to VEGFA and PGE2 between women with PCOS and healthy oocyte donors. Our hypothesis is that ovarian microvascular endothelial cells from women with PCOS exhibit differential angiogenic responses to VEGFA and PGE2 compared to endothelial cells from healthy women. To test this hypothesis, ovarian microvascular endothelial cells were isolated from follicular aspirates obtained during oocyte retrieval from healthy women undergoing ovarian stimulation for oocyte donation (HOMECs, n=4) and those with PCOS undergoing ovarian stimulation for in vitro fertilization (PCOMECs, n=4-6). Migration was quantified by counting cells which migrated through a porous membrane in vitro in response to treatment with either VEGFA (5 ng/ml) or PGE2 (1 µM). Migration is expressed as a percent relative to no treatment (basal). VEGFA increased migration in both HOMECS (199±55%) and PCOMECS (158±13%) when compared to basal migration (basal=100%). PGE2 increased migration in HOMECs (183± 48%). In contrast, PGE2 did not increase migration in PCOMECs (126±21%) above basal migration. mRNA levels for VEGFA receptors (FLT1 and KDR) and PGE2 receptors (PTGER1, PTGER2, PTGER3 and PTGER4) were quantified by qPCR and normalized to GAPDH mRNA in each sample. HOMECs and PCOMECs expressed mRNA for FLT1, KDR, PTGER1, PTGER2, PTGER3, and PTGER4. HOMECs and PCOMECs were found to have similar levels of FLT-1(HOMEC mean 1.78 ± 0.87, PCOMEC means 0.66 ± 0.31) and KDR mRNA (HOMEC mean 1.13 ± 0.29, PCOMEC 1.10 ± 0.49). PCOMECs were found to have significantly higher levels of PTGER2 mRNA (HOMEC mean 1.08 ± 0.23, PCOMEC 3.63 ± 0.99), and significantly lower levels of PTGER3 when compared with HOMECS (HOMEC mean 1.04 ± 0.17, PCOMEC 0.48 ± 0.06). Similar levels of both PTGER1 (HOMEC 1.01±0.10, PCOMEC 0.72±0.18) and PTGER4 mRNA were found in the two groups (HOMEC 1.16±0.29, PCOMEC 0.61±0.14). Overall, HOMECs and PCOMECs both respond to VEGFA as a stimulus for migration; however, only HOMECs increased migration in response to PGE2. These data indicate that altered PGE2 signaling may impair endothelial cell migration in women with PCOS, mediated by either higher PTGER2 and lower PTGER3 in PCOMECs. Dysfunctional endothelial migration may contribute to impaired ovarian follicular growth and/or anovulation and subsequent infertility seen in PCOS. Future studies will explore the functional activity of PTGER2 and PTGER3 receptors and assess additional aspects of endothelial cell function, including proliferation and sprouting, to better understand the role of microvascular impairments in PCOS. The Jones Family Foundation generously funded this project.