FM19G11, a New Hypoxia-inducible Factor (HIF) Modulator, Affects Stem Cell Differentiation Status

• Published in: The Journal of biological chemistry Vol. 285; no. 2; pp. 1333 - 1342
• Main Authors: Moreno-Manzano, Victoria, Rodríguez-Jiménez, Francisco J., Aceña-Bonilla, Jose L., Fustero-Lardíes, Santos, Erceg, Slaven, Dopazo, Joaquin, Montaner, David, Stojkovic, Miodrag, Sánchez-Puelles, Jose M.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 08-01-2010; American Society for Biochemistry and Molecular Biology
• Subjects: Acetylation - drug effects; Adult Stem Cells - cytology; Adult Stem Cells - metabolism; Animals; Basic Helix-Loop-Helix Transcription Factors - antagonists & inhibitors; Basic Helix-Loop-Helix Transcription Factors - metabolism; Benzamides - metabolism; Benzoates - metabolism; Cell Differentiation - drug effects; Cell Hypoxia - drug effects; Cell Hypoxia - physiology; Cell/Differentiation; Development Differentiation/Stem Cell; Diseases/Neurological; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Ependyma - cytology; Ependyma - metabolism; Gene Expression Regulation - drug effects; Gene Expression Regulation - physiology; HeLa Cells; Histones - metabolism; Histones/Acetylase; Homeodomain Proteins - metabolism; Humans; Hypoxia-Inducible Factor 1, alpha Subunit - antagonists & inhibitors; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Male; Molecular Basis of Cell and Developmental Biology; Nanog Homeobox Protein; Octamer Transcription Factor-3 - metabolism; Oxygen/Hypoxia; p300-CBP Transcription Factors - metabolism; Pharmaceutical Preparations; Rats; Rats, Sprague-Dawley; Response Elements - physiology; SOXB1 Transcription Factors - metabolism; Stem Cells; Stem Cells/Neural; Transcription, Genetic - drug effects; Transcription, Genetic - physiology; Transforming Growth Factor alpha - metabolism; Stem Cells; Cell/Differentiation; Development Differentiation/Stem Cell; Oxygen/Hypoxia; Diseases/Neurological; Histones/Acetylase; Stem Cells/Neural
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M109.008326

The biology of the α 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 article show the discovery of FM19G11, a novel chemical entity that inhibits HIFα proteins that repress target genes of the two α subunits, in various tumor cell lines as well as in adult and embryonic stem cell 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 embryonic stem cells, FM19G11 activity occurs in ependymal progenitor stem cells from rats (epSPC), a cell model reported for spinal cord regeneration, which allows the progression of oligodendrocyte cell differentiation in a hypoxic environment, has created interest in its characterization for pharmacological research. Experiments using small interfering RNA showed a significant depletion in Sox2 protein only in the case of HIF2α silencing, but not in HIF1α-mediated ablation. Moreover, chromatin immunoprecipitation data, together with the significant presence of functional hypoxia response element consensus sequences 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 acetyltransferase 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.