title: Hypoxia-inducible factors have distinct and stage-specific roles during reprogramming of human cells to pluripotency.
abstract: Pluripotent stem cells have distinct metabolic requirements, and reprogramming cells to pluripotency requires a shift from oxidative to glycolytic metabolism. Here, we show that this shift occurs early during reprogramming of human cells and requires hypoxia-inducible factors (HIFs) in a stage-specific manner. HIF1α and HIF2α are both necessary to initiate this metabolic switch and for the acquisition of pluripotency, and the stabilization of either protein during early phases of reprogramming is sufficient to induce the switch to glycolytic metabolism. In contrast, stabilization of HIF2α during later stages represses reprogramming, partly because of the upregulation of TNF-related apoptosis-inducing ligand (TRAIL). TRAIL inhibits induced pluripotent stem cell (iPSC) generation by repressing apoptotic caspase 3 activity specifically in cells undergoing reprogramming but not human embryonic stem cells (hESCs), and inhibiting TRAIL activity enhances human iPSC generation. These results shed light on the mechanisms underlying the metabolic shifts associated with the acquisition of a pluripotent identity during reprogramming.
authors.affiliation: Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Mathieu
authors.affiliation: Department of Biology, University of Washington, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Yalan
authors.initials: Y
authors.lastname: Xing
authors.affiliation: Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; Department of Chemistry, University of Washington, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Henrik
authors.initials: H
authors.lastname: Sperber
authors.affiliation: Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Ferreccio
authors.affiliation: Department of Pharmacology, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Agoston
authors.affiliation: Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Kavitha T
authors.initials: KT
authors.lastname: Kuppusamy
authors.affiliation: Department of Pharmacology, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; Howard Hughes Medical Institute, University of Washington, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA.
authors.forename: Randall T
authors.initials: RT
authors.lastname: Moon
authors.affiliation: Department of Biochemistry, University of Washington School of Medicine, Seattle, WA 98109-4714, USA; Department of Biology, University of Washington, Seattle, WA 98109-4714, USA; Department of Chemistry, University of Washington, Seattle, WA 98109-4714, USA; The Institute for Stem Cell and Regenerative Medicine (ISCRM), University of Washington School of Medicine, Seattle, WA 98109-4714, USA. Electronic address: hannele@uw.edu.
authors.forename: Hannele
authors.initials: H
authors.lastname: Ruohola-Baker
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D034741:RNA, Small Interfering; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D000099162:Endothelial PAS Domain-Containing Protein 1
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D053148:Caspase 3; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D034741:RNA, Small Interfering; D053221:TNF-Related Apoptosis-Inducing Ligand; D000099162:Endothelial PAS Domain-Containing Protein 1; C497442:HIF1A protein, human