title: Stimulation of the toll-like receptor 3 promotes metabolic reprogramming in head and neck carcinoma cells.
abstract: In this study, a possible link between the innate immune recognition receptor TLR3 and metabolic reprogramming in Head and Neck carcinoma (HNC) cells was investigated. The effects of TLR3 stimulation/knock-down were assessed under several culture conditions in 4 HNC cell-lines by cell growth assays, targeted metabolomics, and glycolysis assays based on time-resolved analysis of proton release (Seahorse analyzer). The stimulation of TLR3 by its synthetic agonist Poly(A:U) resulted in a faster growth of HNC cells under low foetal calf serum conditions. Targeted analysis of glucose metabolism pathways demonstrated a tendency towards a shift from tricarboxylic acid cycle (Krebs cycle) to glycolysis and anabolic reactions in cells treated with Poly(A:U). Glycolysis assays confirmed that TLR3 stimulation enhanced the capacity of malignant cells to switch from oxidative phosphorylation to extra-mitochondrial glycolysis. We found evidence that HIF-1α is involved in this process: addition of the TLR3 agonist resulted in a higher cell concentration of the HIF-1α protein, even in normoxia, whereas knocking-down TLR3 resulted in a lower concentration, even in hypoxia. Finally, we assessed TLR3 expression by immunohistochemistry in a series of 7 HNSCC specimens and found that TLR3 was detected at higher levels in tumors displaying a hypoxic staining pattern. Overall, our results demonstrate that TLR3 stimulation induces the Warburg effect in HNC cells in vitro, and suggest that TLR3 may play a role in tumor adaptation to hypoxia.
authors: University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France.
authors: Mathieu
authors: M
authors: Veyrat
authors: Equipe 11 Labélisée par la Ligue Nationale Contre le Cancer, INSERM U1138, Centre de Recherche des Cordeliers, Paris, France.|Metabolomics and Molecular Cell Biology Platforms, Gustave Roussy, Villejuif, France.
authors: Sylvère
authors: S
authors: Durand
authors: Department of Pathology, Lariboisière Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France.
authors: Marion
authors: M
authors: Classe
authors: Division of Molecular Medicine, Rudjer Boskovic Institute, Zagreb, Croatia.
authors: Tanja Matijevic
authors: TM
authors: Glavan
authors: University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France.|Department of Head and Neck surgery, Lariboisière Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France.
authors: Natalie
authors: N
authors: Oker
authors: University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France.
authors: Nikiforos-Ioannis
authors: NI
authors: Kapetanakis
authors: University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France.
authors: Xiaojun
authors: X
authors: Jiang
authors: University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France.
authors: Aurore
authors: A
authors: Gelin
authors: Department of Head and Neck surgery, Lariboisière Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France.
authors: Philippe
authors: P
authors: Herman
authors: Department of Biopathology, Gustave Roussy, Villejuif, France.
authors: Odile
authors: O
authors: Casiraghi
authors: Department of Neuropathology, New York University School of Medicine, New York, NY, USA.
authors: David
authors: D
authors: Zagzag
authors: Equipe 11 Labélisée par la Ligue Nationale Contre le Cancer, INSERM U1138, Centre de Recherche des Cordeliers, Paris, France.|Metabolomics and Molecular Cell Biology Platforms, Gustave Roussy, Villejuif, France.
authors: David
authors: D
authors: Enot
authors: University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France.
authors: Pierre
authors: P
authors: Busson
authors: University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France.|Department of Head and Neck surgery, Lariboisière Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France.
authors: Benjamin
authors: B
authors: Vérillaud
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism; D000071231:Carbonic Anhydrase IX / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D065150:Cellular Reprogramming / Q000187:drug effects*; D002952:Citric Acid Cycle / Q000187:drug effects; D004734:Energy Metabolism / Q000187:drug effects*; D006019:Glycolysis / Q000187:drug effects; D006258:Head and Neck Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D010085:Oxidative Phosphorylation / Q000187:drug effects; D011063:Poly A-U / Q000494:pharmacology*; D034622:RNA Interference; D015398:Signal Transduction / Q000187:drug effects; D000077195:Squamous Cell Carcinoma of Head and Neck; D013997:Time Factors; D051196:Toll-Like Receptor 3 / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D000072258:Tumor Hypoxia
chemical_list: D000951:Antigens, Neoplasm; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D011063:Poly A-U; C089226:CA9 protein, human; D000071231:Carbonic Anhydrase IX
keywords: HIF; innate immunity; metabolomics; toll-like receptor 3; warburg effect