Abstract

The biology of the alpha subunits of Hypoxia Inducible Factors (HIFα) has expanded from their role in angiogenesis to their current position in the self-renewal and differentiation of stem cells. The results reported in this paper show the discovery of FM19G11, a novel chemical entity that inhibits the HIFα proteins that repress the target genes of the two α subunits, in various tumor cell lines as well as in adult and embryonic stem cell (ESC) models from rodents and humans, respectively. FM19G11 inhibits at nanomolar range the transcriptional and protein expression of Oct4, Sox2, Nanog and Tgf-α undifferentiating factors, in adult rat and human ESC. FM19G11 activity occurs in ependymal progenitor stem cells from rats (epSPC) - a cell model reported for spinal cord regeneration. That it allows the progression of oligodendrocyte cell differentiation in a hypoxic environment has created interest in its characterization for pharmacological research. Experiments using siRNA showed a significant depletion in Sox2 protein only in the case of HIF2α silencing, but not in HIF1α-mediated ablation. Moreover, ChIP data, together with the significant presence of functional hypoxia response element consensus sequences (HREs) in the promoter region of Sox2, strongly validated that this factor behaves as a target gene of HIF2α in epSPCs. FM19G11 causes a reduction of overall histone acetylation with significant repression of p300, a histone acetyl transferase required as a co-factor for HIF-transcription activation. Arrays carried out in the presence and absence of the inhibitor showed the predominant involvement of epigenetic associated events mediated by the drug.