Effect of GFI1B ectopic expression in the hematopoietic or endothelial commitment of human iPSC derived CD34+CD43-CD73- hemogenic endothelium (HE)
In this study we focused on unrevealing the role of major transcriptional factor GFI1B and its cofactor, LSD1 in human endothelial to hematopoietic transition (EHT). We applied irreversible LSD1 inhibitor (GSK-LSD1) to healthy iPSC lines. Interestingly, LSD1 inhibited healthy lines which showed complete absence of hematopoietic cell output, did not showed detection of GFI1B expression, suggesting a timed transcriptional program. In order to test this hypothesis, we ectopically expressed GFI1B in the uncommitted HE cells, leading to downregulation of endothelial genes and an upregulation of hematopoietic genes, including GATA2, KIT, RUNX1 and SPI1. Overall design: To investigate this, we enriched CD34+ HE cells from iPSC differentiation. We lentivirally transduced these cells with empty vector (EV) or GFI1B, and harvested them 3 days after transduction for RNA seq analysis. We had three replicates for each condition