E-Poster Presentation ESA-SRB-ANZBMS 2021

Proteomes of endometrial stromal cell-derived small EVs indicate decidualisation potential (#594)

Shanti Gurung 1 2 , David Greening 3 4 5 6 , Alin Rai 3 5 6 , Qi Hui Poh 3 4 , Jemma Evans 1 7 8 , Lois Salamonsen 1 9
  1. Centre for Reproductive Health/Hudson Institute of Medical Research/Monash University, Clayton, VIC, Australia
  2. The Ritchie Centre, Hudson Institute of Medical Research, Clayton, VIC, Australia
  3. Baker Heart and Diabetes Institute, Molecular Proteomics, Melbourne, Victoria, Australia
  4. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University,, Bundoora, Victoria, Australia
  5. Central Clinical School, Monash University, Melbourne, Victoria, Australia
  6. Baker Department of Cardiometabolic Health, University of Melbourne, Melbourne, Victoria, Australia
  7. Department of Physiology, Monash University , Clayton, Victoria, Australia
  8. Nucleus Network, Melbourne, Victoria, Australia
  9. Department of Molecular and Translational Medicine , Hudson Institute of Medical Research, Clayton, Victoria, Australia

Background: Proper decidualisation is vital in preparation for endometrial health including, embryo receptivity, controlled cytotrophoblast invasion and subsequent endometrial regeneration. Aim: To investigate the role of small extracellular vesicle (sEV) mediated communication during decidualisation.

Methods: Human endometrial stromal cells (SC) were treated with estrogen and progesterone for 14 days (d) and grouped as well (WD) and poorly decidualised (PD) based on secreted prolactin level. sEV were isolated from conditioned media using differential centrifugation and subjected to mass spectrometry-based quantitative proteomic analysis.

Results and conclusions: On d2 PD- versus WD-SC-sEVs; 58 proteins were differentially regulated (DE) with 17 down-regulated (involved in complement/coagulation cascades, platelet degranulation and fibrinolysis) and 39 up-regulated (involved in focal adhesion, glycolysis /gluconeogenesis, PI3K-Akt signaling pathway and leukocyte transendothelial migration). On d14, in PD- versus WD-SC-sEVs, FLNA was down-regulated while 21 proteins were up-regulated associated with complement/coagulation cascades (C3, C6), platelet degranulation (SERPINA4, ITIH4), B-cell receptor signaling and innate immune response (immunoglobulins). Six proteins, PRDX1, PFN1 VCL, RAC1, THBS4 and ANXA1 (for invasion, migration and embryo implantation) were uniquely present in WD-sEVs. Interrogation of changes from d2 to d14 identified no significant changes in WD-SC-sEVs, however, in PD-SC-sEVs, proteins involved in complement and coagulation cascade were significantly upregulated at d14 while 36 proteins (involved in focal adhesion, regulation of actin cytoskeleton and glycolysis/gluconeogenesis were only present in d2.

Our findings provide an insight into sEVs as a mode of cellular communication by SC, provides insight into sEVs-proteomes as a benchmark of well decidualised SC and how failure of appropriate decidualisation may lead to dysregulated communication that is critical for embryo implantation, enabling and limiting trophoblast invasion during placentation and sensing a healthy embryo.