Pluto Bioinformatics

GSE141773: Wnt / Beta-Catenin signalling controls epiblast self-organization in naive and paused states of pluripotency

Bulk RNA sequencing

ES cells grown in growth factor depleted matrigel could recapitulate the morphogenesis of the epiblast from E4.5 to E5.5. This process is characterized with the establishment of epithelial polarity. We used Next-generation sequencing (NGS) technique to analyze the transcriptional change of ES cells grown in 3D culture conditions in the presence of DMSO, 2i, CH or FGF2/Activin. The comparison of transcriptom between different conditions helps us to understand the molecular mechanism underlying morphogenesis of the epiblast. We found nave pluripotency factors are kept in a high level in 2i and CH treatment in comparison with DMSO and FGF2/Activin treatment. We identified 5 nave pluripotency factors which may have potential regulatory function in epithelial polarity. To further analyze these 5 pluripotency factors, we found Esrrb is the core transcriptional factor countering epithelial polarity. We also used Next-generation sequencing (NGS) technique to analyze the transcriptional change of Esrrb delta/delta (+4OHT) and control Esrrb fl/fl (-4OHT) ES cells in the presence of CH. Esrrb delta/delta (+4OHT) cell grown in 3D become polarized at 24 hours, whereas the control Esrrb fl/fl (-4OHT) ES cells are still non-polarized. By analyzing the RNA-Seq data, we found the enrichment of MAPK, Apoptosis, Focal adhesion and Tight junction KEGG pathways in Esrrb delta/delta compared to control Esrrb fl/fl cells. This result suggests the essential function of Esrrb in regulating epithelial polarity. SOURCE: Jie Wu (jie.wu@dcb.unibe.ch) - Department of Chemistry and Biochemistry