Oral Presentation ESA-SRB-APEG-NZSE 2022

Infertile human endometrial organoid apical protein secretions are dysregulated and impair trophoblast progenitor cell adhesion (#53)

Wei Zhou 1 2 , Siena Barton 1 2 , JinWei Cui 3 , Leiani L Santos 1 2 , Guannan Yang 1 2 , Kate Stern 1 4 5 , Violet Kieu 1 4 5 , Wan Tinn Teh 1 4 5 6 , Catarina Ang 1 4 , Tarana Lucky 4 7 , Joseph Sgroi 5 6 , Louie Ye 1 , Dimitriadis Evdokia 1 2
  1. Department of Obstetrics, Gynaecology and Newborn Health, University of Melbourne, Melbourne, VIC, Australia
  2. Gynaecology Research Centre, Royal Women's Hospital, Melbourne, VIC, Australia
  3. Priority Research Centre for Reproductive Science, School of Environmental and Life Sciences, University of Newcastle, Newcastle, New South Wales, Australia
  4. The Royal Women’s Hospital, Parkville, VIC, Australia, Melbourne, Victoria, Australia
  5. Melbourne IVF, Melbourne, Victoria
  6. Epworth HealthCare, Melbourne, Victoria, Australia
  7. School of Medicine, Griffith University, Gold Coast, Queensland, Australia

Embryo implantation failure leads to infertility and remains a challenging problem for IVF. As an important approach to regulate implantation, endometrial glands produce and secrete factors apically into the uterine cavity in the receptive phase to prepare the initial blastocyst adhesion and implantation. Organoids were recently developed from human endometrial glands and show long-term expandability, genetic stability and maintenance of their hormone responsiveness. Importantly, organoid exhibits similar apical-basal polarity compared to endometrial gland making it an ideal model to study glandular secretions. We established organoids using endometrial biopsies from women with normal fertility and primary infertility (referred to as fertile and infertile organoids). Organoids from both groups were treated with hormones to model the receptive phase of the endometrial glands and intra-organoid apical fluid (IOF) was collected to compare the apical protein secretion profile. Our data show that infertile organoids were dysregulated in their response to estrogen and progesterone treatment. Proteomic analysis of IOF identified 131 decreased and 19 increased proteins in infertile group compared to fertile (>1.5-fold change). Many of the proteins were similarly differentially regulated in organoid cells at the mRNA level. To determine the effect of organoid apical secretion on blastocyst adhesion, we first developed epithelial monolayers using fertile organoids and compared them with previously established primary human endometrial epithelial monolayers. Using miR-29c as an example, we confirmed that both models respond similarly to microRNA overexpression. IOF was collected after hormone stimulation to treat trophoblast progenitor spheroids (blastocyst surrogates) and their adhesion on the organoid-derived endometrial cell monolayer determined. Incubation of infertile IOF significantly reduced trophoblast spheroid adhesive capacity compared to fertile IOF (P < 0.0001) and medium control (P < 0.01), respectively. Together, this study paves the way to determine the molecular mechanisms by which endometrial glandular apical released factors regulate blastocyst initial attachment.