Metabolic stress is a barrier to Epstein–Barr virus-mediated B-cell immortalization

• Published in: Proceedings of the National Academy of Sciences - PNAS 2016-02, Vol.113 (6), p.E782-E790
• Main Authors: McFadden, Karyn, Hafez, Amy Y., Kishton, Rigel, Messinger, Joshua E., Nikitin, Pavel A., Rathmell, Jeffrey C., Luftig, Micah A.
• Format: Article
• Language: English
• Subjects: Autophagy - drug effects; B-Lymphocytes - drug effects; B-Lymphocytes - pathology; B-Lymphocytes - ultrastructure; B-Lymphocytes - virology; Biological Sciences; Cell cycle; Cell Cycle Checkpoints - drug effects; Cell Proliferation - drug effects; Cell Respiration - drug effects; Cell Transformation, Viral - drug effects; Cells; Cellular Senescence - drug effects; Deoxyglucose - pharmacology; Dimethylformamide - pharmacology; DNA damage; DNA Damage - drug effects; DNA Replication - drug effects; Epstein-Barr virus; Herpes viruses; Herpesviridae; Herpesvirus; Herpesvirus 4, Human - drug effects; Herpesvirus 4, Human - physiology; Humans; Mechanistic Target of Rapamycin Complex 1; Metabolism; Metabolomics; Mitochondria - drug effects; Mitochondria - metabolism; Multiprotein Complexes - metabolism; Oncogenes; Phosphorylation; PNAS Plus; Signal transduction; Signal Transduction - drug effects; Stress, Physiological - drug effects; TOR Serine-Threonine Kinases - metabolism; Transcription Factors - metabolism; Transcriptome - genetics; Tumor Suppressor Protein p53 - metabolism
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1517141113

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.