title: Bach1 differentially regulates distinct Nrf2-dependent genes in human venous and coronary artery endothelial cells adapted to physiological oxygen levels.
abstract: The effects of physiological oxygen tension on Nuclear Factor-E2-Related Factor 2 (Nrf2)-regulated redox signaling remain poorly understood. We report the first study of Nrf2-regulated signaling in human primary endothelial cells (EC) adapted long-term to physiological O2 (5%). Adaptation of EC to 5% O2 had minimal effects on cell ultrastructure, viability, basal redox status or HIF1-α expression. Affymetrix array profiling and subsequent qPCR/protein validation revealed that induction of select Nrf2 target genes, HO-1 and NQO1, was significantly attenuated in cells adapted to 5% O2, despite nuclear accumulation and DNA binding of Nrf2. Diminished HO-1 induction under 5% O2 was stimulus independent and reversible upon re-adaptation to air or silencing of the Nrf2 repressor Bach1, notably elevated under 5% O2. Induction of GSH-related genes xCT and GCLM were oxygen and Bach1-insensitive during long-term culture under 5% O2, providing the first evidence that genes related to GSH synthesis mediate protection afforded by Nrf2-Keap1 defense pathway in cells adapted to physiological O2 levels encountered in vivo.
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Chapple
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Keeley
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Daniela
authors.initials: D
authors.lastname: Mastronicola
authors.affiliation: Genomics Centre, King's College London, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Matthew
authors.initials: M
authors.lastname: Arno
authors.affiliation: Centre for Ultrastructural Imaging, King's College London, London SE1 9NH, UK.
authors.forename: Gema
authors.initials: G
authors.lastname: Vizcay-Barrena
authors.affiliation: Centre for Ultrastructural Imaging, King's College London, London SE1 9NH, UK.
authors.forename: Roland
authors.initials: R
authors.lastname: Fleck
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London SE1 9NH, UK.
authors.forename: Richard C M
authors.initials: RCM
authors.lastname: Siow
authors.affiliation: Cardiovascular Division, British Heart Foundation Centre of Research Excellence, Faculty of Life Sciences & Medicine, King's College London, 150 Stamford Street, London SE1 9NH, UK. Electronic address: giovanni.mann@kcl.ac.uk.
authors.forename: Giovanni E
authors.initials: GE
authors.lastname: Mann
mesh_terms: D027182:Amino Acid Transport System y+ / Q000378:metabolism; D000975:Antioxidants / Q000378:metabolism; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics* / Q000378:metabolism; D003331:Coronary Vessels / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D051856:Fanconi Anemia Complementation Group Proteins / Q000235:genetics* / Q000378:metabolism; D005721:Glutamate-Cysteine Ligase / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D051547:Heme Oxygenase-1 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D000072019:Kelch-Like ECH-Associated Protein 1 / Q000235:genetics / Q000378:metabolism*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000235:genetics / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics* / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D014680:Veins / Q000378:metabolism
chemical_list: D027182:Amino Acid Transport System y+; D000975:Antioxidants; C102108:BACH1 protein, human; D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D051856:Fanconi Anemia Complementation Group Proteins; 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; C495635:NFE2L2 protein, human; D017382:Reactive Oxygen Species; C118895:SLC7A11 protein, human; D051547:Heme Oxygenase-1; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; C546365:GCLM protein, human; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione; D010100:Oxygen
keywords: Bach1; Coronary artery; Endothelial cells; GCL; Glutathione; Glutathione reductase GR; HO-1; Mitochondria; NQO1; Normoxia; Nuclear Factor-E2-Related Factor 2, Nrf2; Physiological oxygen tension; Redox signaling; Sequestosome-1; Solute Carrier Family 7-anionic amino acid transporter light chain xCT; Thioredoxin reductase-1