Effect of Ulipristal Acetate (UPA) on the gene expression profiling of endometrial cells in culture

COHEN DJ; LUIS BAHAMONDE; CUASNICU PS; GARCIA A; COTAN D; MUNUCE MJ; HORCAJADAS J; CAILLE, A; JIMENEZ GUERRERO M

Congreso; 34th Annual Meeting ESHRE; 2018

Study question:To assess the effect ofUPA, in concentrations compatible with emergency contraception (EC), on geneprofile expression of an endometrium cell line in culture.Summary answer:The presence of UPAmodifies the transcriptomic signature of endometrial cells in culture.What is known already:Emergency contraceptionconsists in the use of drugs or devices to prevent pregnancy after unprotectedsexual intercourse. Considering the fundamental role that progesterone hasthroughout the process ranging from ovulation to implantation and embryodevelopment, UPA has been developed as a selective progesterone receptormodulator (SPRM) with agonist/antagonist activity and pharmacologicalapplications in EC.The principal mechanismof action of UPA is the inhibition or delay of follicular rupture whenadministered previous to LH peak. However, given its high effectiveness in theprevention of pregnancy (up to 5 days after intercourse), post-ovulatoryeffects related to endometrial receptivity, cannot be excluded.Study design, size, duration:This basic researchwork was conducted in 2017 and designed as a prospective study. Epithelialcells of human endometrium carcinoma cell line (HEC-1A) were exposed 24 and 48h to UPA (100, 250, 500 ng/ml) in triplicates. Total RNA was extracted and 192genes associated with endometrial receptivity were evaluated by quantitativePCR.Participants/materials, setting, methods:Fluidigm Microfluidictechnology combines microarrays with real time PCR, where 192 genes areanalyzed. This selected panel was analyzed after 24 and 48 h in culture. Fromnormalized Cqs with respect to the constituent genes, Fold-Change (FC ≥ 0,5 inabsolute value) was calculated to determine the differential gene expressionbetween samples treated with UPA and controls. Gene Ontology was performed toidentify the biological function of those representative genes.Main results and the role of chance:Data showed that by 24h of incubation, there are no significant changes of transcriptomic profile inthose cells exposed to UPA (100, 250, 500 ng/mL) respect to controls culturedwithout the drug. However, by 48 h of incubation, there is a significant(p<0,05) increase in total transcriptomic profile in those cells exposed toUPA (at all concentration assayed) compared to controls. To determine whichgenes are differentially expressed after 48 h of culture, FC of the 192 genesrelated to endometrial receptivity were analyzed. Absolute value of FCincreases with the concentration of UPA in culture, suggesting a dose-dependenttrend of down-regulation of gene expression in the panel of genes analyzed.Considering those genes with FC ≥ 0,5 in absolute value, a Gene Ontology(DAVID) was performed to identify the biological function of thoserepresentative genes as well as the possible effects of UPA on those genes.Genes related to cellular adhesion and proliferation, signal transduction,growth factors, cytokine activity and estradiol response seem to be downregulated after UPA exposure.Limitations, reasons for caution:Our study was carriedout in vitro with a human endometrium carcinoma cellular model that expressesestrogen, progesterone, molecular adhesion receptors and proteases This couldlimit the extrapolation of results to the UPA effects on endometrial tissue.Wider implications of the findings:The present studysuggests that there are changes in gene expression of endometrial cells exposedto UPA at concentrations compatible with EC. Our data brings new evidence forthe study of the molecular mechanisms of action of UPA, used for contraceptiveproposal, on endometrium receptivity associated with embryo implantation.Trial registration number:This study was supported bySINAE, University of Rosario (BIO 486 Res. 1480/2016), São Paulo ResearchFoundation (FAPESP) 2015/20504-9, CONICET and ANPCyT (PICT 2016-1057).