title: Metabolic stress is a barrier to Epstein-Barr virus-mediated B-cell immortalization.
abstract: Epstein-Barr virus (EBV) is an oncogenic herpesvirus that has been causally linked to the development of B-cell and epithelial malignancies. Early after infection, EBV induces a transient period of hyperproliferation that is suppressed by the activation of the DNA damage response and a G1/S-phase growth arrest. This growth arrest prevents long-term outgrowth of the majority of infected cells. We developed a method to isolate and characterize infected cells that arrest after this early burst of proliferation and integrated gene expression and metabolic profiling to gain a better understanding of the pathways that attenuate immortalization. We found that the arrested cells have a reduced level of mitochondrial respiration and a decrease in the expression of genes involved in the TCA cycle and oxidative phosphorylation. Indeed, the growth arrest in early infected cells could be rescued by supplementing the TCA cycle. Arrested cells were characterized by an increase in the expression of p53 pathway gene targets, including sestrins leading to activation of AMPK, a reduction in mTOR signaling, and, consequently, elevated autophagy that was important for cell survival. Autophagy was also critical to maintain early hyperproliferation during metabolic stress. Finally, in assessing the metabolic changes from early infection to long-term outgrowth, we found concomitant increases in glucose import and surface glucose transporter 1 (GLUT1) levels, leading to elevated glycolysis, oxidative phosphorylation, and suppression of basal autophagy. Our study demonstrates that oncogene-induced senescence triggered by a combination of metabolic and genotoxic stress acts as an intrinsic barrier to EBV-mediated transformation.
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for Virology, Duke University School of Medicine, Durham, NC 27710;
authors.forename: Karyn
authors.initials: K
authors.lastname: McFadden
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for Virology, Duke University School of Medicine, Durham, NC 27710;
authors.forename: Amy Y
authors.initials: AY
authors.lastname: Hafez
authors.affiliation: Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, NC 27710;
authors.forename: Rigel
authors.initials: R
authors.lastname: Kishton
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for Virology, Duke University School of Medicine, Durham, NC 27710;
authors.forename: Joshua E
authors.initials: JE
authors.lastname: Messinger
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for Virology, Duke University School of Medicine, Durham, NC 27710;
authors.forename: Pavel A
authors.initials: PA
authors.lastname: Nikitin
authors.affiliation: Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, NC 27710; Department of Immunology, Duke University School of Medicine, Durham, NC 27710; Duke Molecular Physiology Institute, Duke University, Durham, NC 27710.
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Rathmell
authors.affiliation: Department of Molecular Genetics and Microbiology, Center for Virology, Duke University School of Medicine, Durham, NC 27710; micah.luftig@duke.edu.
authors.forename: Micah A
authors.initials: MA
authors.lastname: Luftig
mesh_terms: D001343:Autophagy / Q000187:drug effects; D001402:B-Lymphocytes / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure / Q000821:virology*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D019069:Cell Respiration / Q000187:drug effects; D002472:Cell Transformation, Viral / Q000187:drug effects; D016922:Cellular Senescence / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects; D004261:DNA Replication / Q000187:drug effects; D003847:Deoxyglucose / Q000494:pharmacology; D004126:Dimethylformamide / Q000494:pharmacology; D004854:Herpesvirus 4, Human / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D000076222:Mechanistic Target of Rapamycin Complex 1; D055432:Metabolomics; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D046912:Multiprotein Complexes / Q000378:metabolism; D009857:Oncogenes; D015398:Signal Transduction / Q000187:drug effects; D013312:Stress, Physiological / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D059467:Transcriptome / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D046912:Multiprotein Complexes; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D004126:Dimethylformamide; D003847:Deoxyglucose; D000076222:Mechanistic Target of Rapamycin Complex 1; D058570:TOR Serine-Threonine Kinases
keywords: B cell; Epstein–Barr virus; autophagy; metabolism; oncogene-induced senescence