Oral Virtual Presentation (Virtual only) ESA-SRB-ANZBMS 2021

Detrimental actions of obesity-associated Advanced Glycation Endproducts on endometrial epithelial cell proliferation are alleviated by antioxidants and are donor-dependent in human endometrial organoids (#186)

Jennifer C Hutchison 1 2 , Jemma Evans 2 , Tracey Edgell 1 2 , Guiying Nie 2 3 , David K Gardner 4 , Lois A Salamonsen 1 2
  1. Department of Molecular and Translational Science, Monash University, Clayton, VIC, Australia
  2. Hudson Institute of Medical Research, Clayton, VIC, Australia
  3. Implantation and Pregnancy Research Laboratory, School of Health and Biomedical Sciences, RMIT University, Bundoora, Vic, Australia
  4. School of Biosciences, University of Melbourne, Melbourne, Victoria, Australia

Background:

Global levels of obesity are rising, with 39% of people overweight or obese (WHO). Obese women experience reduced fertility and increased pregnancy risks including preeclampsia. Advanced Glycation Endproducts (AGEs), a proinflammatory modification of proteins exposed to sugars, are elevated in the uterine fluid of obese women versus lean. AGEs compromise both preimplantation embryo development and endometrial cell functions.

Aim: To investigate therapeutics to restore endometrial cell function, and characterise the effect of obesity-associated AGEs on human endometrial epithelial cell organoids (hEEO).

Methods:

Endometrial epithelial cell line (ECC-1) and hEEO were cultured in AGEs equimolar with lean (2 umol/mol lysine) and obese (8 umol/mol lysine) uterine environments. Real time cell analysis (xCelligence) of ECC-1 examined remedial effects of i) 100 uM metformin; ii) antioxidants (10 uM N-acetyl-cysteine, 10 uM N-acetyl-L-carnitine, 5 uM α-lipoic acid); iii) 25 nM RAGE antagonist (FPS-ZM1). hEEO-derived primary epithelial cells exposed to obese AGEs were examined by xCelligence. Multiplex analysis of chemokine & cytokine secretion (inflammatory determinants) in hEEO conditioned medium. CXCL16 profiled by Luminex analysis and glucose by Randox Daytona analyser in uterine fluid of women undergoing IVF.

Results:

Obese AGEs-reduced ECC-1 proliferation (P<0.001) which was successfully restored by antioxidants. hEEO were functionally impacted by obese AGEs, demonstrating a donor-dependent effect on proliferation. AGEs increased secretion of proinflammatory factors associated with poor pregnancy outcomes, including CXCL16 (P=0.04). Uterine fluid CXCL16 correlated positively to BMI (R=0.26; P=0.02) and uterine glucose (AGEs precursor; R=0.74, P<0.0001), but was not significantly different between IVF cycles resulting in pregnancy vs no pregnancy (P=0.46), nor live birth vs miscarriage (P=0.27).

Conclusion:

AGEs promote a uterine inflammatory milieu hostile to implantation. Antioxidants alleviate the effects of AGEs on ECC1 cells, providing a potential therapeutic for obese women. Clinically, reduced uterine AGEs may improve fertility for obese women wishing to conceive.