title: NRF2 Orchestrates the Metabolic Shift during Induced Pluripotent Stem Cell Reprogramming.
abstract: The potential of induced pluripotent stem cells (iPSCs) in disease modeling and regenerative medicine is vast, but current methodologies remain inefficient. Understanding the cellular mechanisms underlying iPSC reprogramming, such as the metabolic shift from oxidative to glycolytic energy production, is key to improving its efficiency. We have developed a lentiviral reporter system to assay longitudinal changes in cell signaling and transcription factor activity in living cells throughout iPSC reprogramming of human dermal fibroblasts. We reveal early NF-κB, AP-1, and NRF2 transcription factor activation prior to a temporal peak in hypoxia inducible factor α (HIFα) activity. Mechanistically, we show that an early burst in oxidative phosphorylation and elevated reactive oxygen species generation mediates increased NRF2 activity, which in turn initiates the HIFα-mediated glycolytic shift and may modulate glucose redistribution to the pentose phosphate pathway. Critically, inhibition of NRF2 by KEAP1 overexpression compromises metabolic reprogramming and results in reduced efficiency of iPSC colony formation.
authors.affiliation: Stem Cell Group, Cardiovascular and Cell Sciences Research Institute, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK. Electronic address: rmjbkeh@ucl.ac.uk.
authors.forename: Kate E
authors.initials: KE
authors.lastname: Hawkins
authors.affiliation: Stem Cell Group, Cardiovascular and Cell Sciences Research Institute, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK; Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Shona
authors.initials: S
authors.lastname: Joy
authors.affiliation: Stem Cell Group, Cardiovascular and Cell Sciences Research Institute, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK; Wits/SAMRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg 2000, South Africa; Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK.
authors.forename: Juliette M K M
authors.initials: JM
authors.lastname: Delhove
authors.affiliation: Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Vassilios N
authors.initials: VN
authors.lastname: Kotiadis
authors.affiliation: Institute of Functional Biology and Genomics, University of Salamanca-CSIC, 37007 Salamanca, Spain; Institute of Biomedical Research of Salamanca, University Hospital of Salamanca, 37007 Salamanca, Spain.
authors.forename: Emilio
authors.initials: E
authors.lastname: Fernandez
authors.affiliation: Stem Cell Group, Cardiovascular and Cell Sciences Research Institute, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK; School of Healthcare Science, John Dalton Building, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK.
authors.forename: Lorna M
authors.initials: LM
authors.lastname: Fitzpatrick
authors.affiliation: William Harvey Research Institute, Charterhouse Square, Queen Mary University of London, London EC1M 6BQ, UK.
authors.forename: James R
authors.initials: JR
authors.lastname: Whiteford
authors.affiliation: William Harvey Research Institute, Charterhouse Square, Queen Mary University of London, London EC1M 6BQ, UK.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: King
authors.affiliation: Institute of Functional Biology and Genomics, University of Salamanca-CSIC, 37007 Salamanca, Spain; Institute of Biomedical Research of Salamanca, University Hospital of Salamanca, 37007 Salamanca, Spain.
authors.forename: Juan P
authors.initials: JP
authors.lastname: Bolanos
authors.affiliation: Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Duchen
authors.affiliation: Wits/SAMRC Antiviral Gene Therapy Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg 2000, South Africa; Gene Transfer Technology Group, Institute for Women's Health, University College London, 86-96 Chenies Mews, London WC1E 6HX, UK.
authors.forename: Simon N
authors.initials: SN
authors.lastname: Waddington
authors.affiliation: Stem Cell Group, Cardiovascular and Cell Sciences Research Institute, St. George's University of London, Cranmer Terrace, London SW17 0RE, UK; School of Healthcare Science, John Dalton Building, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK. Electronic address: t.mckay@mmu.ac.uk.
authors.forename: Tristan R
authors.initials: TR
authors.lastname: McKay
mesh_terms: D065150:Cellular Reprogramming; D020405:Dermis / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D006019:Glycolysis / Q000235:genetics; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D000072019:Kelch-Like ECH-Associated Protein 1 / Q000235:genetics* / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics* / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D010085:Oxidative Phosphorylation; D010427:Pentose Phosphate Pathway / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D051267:NF-E2-Related Factor 2; D016328:NF-kappa B; C495635:NFE2L2 protein, human; D017382:Reactive Oxygen Species; D018808:Transcription Factor AP-1; D008156:Luciferases