Toll-Like Receptor 4–Induced Glycolytic Burst in Human Monocyte

Toll-Like Receptor 4–Induced Glycolytic Burst in Human Monocyte

Toll-like Receptor 4−Induced Glycolytic Burst in Human Monocyte-Derived Dendritic Cells Results from p38-Dependent Stabilization of HIF-1α and Increased This information is current as Hexokinase II Expression of September 28, 2021. Laure Perrin-Cocon, Anne Aublin-Gex, Olivier Diaz, Christophe Ramière, Francesco Peri, Patrice André and Vincent Lotteau J Immunol published online 23 July 2018 Downloaded from http://www.jimmunol.org/content/early/2018/07/21/jimmun ol.1701522 http://www.jimmunol.org/ Supplementary http://www.jimmunol.org/content/suppl/2018/07/21/jimmunol.170152 Material 2.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision by guest on September 28, 2021 • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2018 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published July 23, 2018, doi:10.4049/jimmunol.1701522 The Journal of Immunology Toll-like Receptor 4–Induced Glycolytic Burst in Human Monocyte-Derived Dendritic Cells Results from p38-Dependent Stabilization of HIF-1a and Increased Hexokinase II Expression Laure Perrin-Cocon,* Anne Aublin-Gex,* Olivier Diaz,* Christophe Ramie`re,* Francesco Peri,† Patrice Andre´,* and Vincent Lotteau* Cell metabolism now appears as an essential regulator of immune cells activation. In particular, TLR stimulation triggers metabolic reprogramming of dendritic cells (DCs) with an increased glycolytic flux, whereas inhibition of glycolysis alters their functional activation. The molecular mechanisms involved in the control of glycolysis upon TLR stimulation are poorly understood for human Downloaded from DCs. TLR4 activation of human monocyte-derived DCs (MoDCs) stimulated glycolysis with an increased glucose consumption and lactate production. Global hexokinase (HK) activity, controlling the initial rate-limiting step of glycolysis, was also increased. TLR4- induced glycolytic burst correlated with a differential modulation of HK isoenzymes. LPS strongly enhanced the expression of HK2, whereas HK3 was reduced, HK1 remained unchanged, and HK4 was not expressed. Expression of the other rate-limiting glycolytic enzymes was not significantly increased. Exploring the signaling pathways involved in LPS-induced glycolysis with various specific a inhibitors, we observed that only the inhibitors of p38–MAPK (SB203580) and of HIF-1 DNA binding (echinomycin) reduced http://www.jimmunol.org/ both the glycolytic activity and production of cytokines triggered by TLR4 stimulation. In addition, LPS-induced HK2 expression required p38-MAPK–dependent HIF-1a accumulation and transcriptional activity. TLR1/2 and TLR2/6 stimulation increased glucose consumption by MoDCs through alternate mechanisms that are independent of p38–MAPK activation. TBK1 contributed to glycolysis regulation when DCs were stimulated via TLR2/6. Therefore, our results indicate that TLR4-dependent upregulation of glycolysis in human MoDCs involves a p38-MAPK–dependent HIF-1a accumulation, leading to an increased HK activity supported by enhanced HK2 expression. The Journal of Immunology, 2018, 201: 000–000. oll-like receptors are common sensors expressed by den- stimulate naive T cells (2). TLR4 is a receptor for Gram-negative by guest on September 28, 2021 dritic cells (DCs) recognizing pathogen-associated molec- bacteria LPS (3). This receptor can also be activated by other T ular patterns and damage-associated molecular patterns, microbial ligands such as viral proteins (4–6) and by damage- initiating an appropriate immune response to fight infection (1). associated molecular patterns such as high-mobility group box 1 TLR signaling triggers human DC maturation, promoting the se- (HMGB) (7) or oxidized phospholipids (8). TLR4 is also involved cretion of proinflammatory cytokines and inducing the expression in the sensing of nutrients and metabolic stress. Indeed, saturated of costimulatory molecules so that DCs acquire the ability to free fatty acids can activate TLR4 signaling via their binding to the hepatokine Fetuin A, which is a TLR4 ligand (9). Its increased production in steatotic and inflamed liver contributes to the secre- *Centre International de Recherche en Infectiologie, Biologie Cellulaire des Infections tion of inflammatory cytokines by monocytes and adipose tissue Virales, INSERM, U1111, Universite´ Claude Bernard Lyon 1, CNRS, UMR5308, E´cole (10). By regulating the expression of many genes involved in innate Normale Supe´rieure de Lyon, Hospices Civils de Lyon, Universite´ de Lyon, Lyon, France; and †Department of Biotechnology and Biosciences, University of Milano-Bicocca, 20126 immunity and metabolic reprogramming (11), TLR4 is at the Milano, Italy crossroads of innate immunity and metabolic inflammation (12) and ORCIDs: 0000-0003-0501-6733 (L.P.-C.); 0000-0002-1981-7155 (C.R.); 0000-0002- is involved in the pathogenesis of metabolic diseases such as obesity 5834-7395 (P.A.). and type 2 diabetes (13). Received for publication November 10, 2017. Accepted for publication July 3, 2018. Glucose is a major nutrient for cellular bioenergetics production. This work was supported by European Commission–granted TOLLerant Project Glycolysis converts glucose to pyruvate by a series of enzymatic H2020-MSC-ETN-642157 (to F.P.), INSERM Ebola Task Force I3M, and Fondation reaction steps, generating 2 mol of ATP per mole of glucose. In- pour la Recherche Me´dicale Grant DEQ20160334893 (to V.L.). termediary metabolites of glycolysis are precursors also fueling the Address correspondence and reprint requests to Dr. Laure Perrin-Cocon and Dr. Vincent Lotteau, Inserm U1111 - Centre International de Recherche en Infectio- pentose phosphates, lipids, and amino acid pathways (11). Three rate- logie, 21 Avenue Tony Garnier, 69365 Lyon Cedex 7, France. E-mail addresses: limiting enzymes are controlling the glycolytic flux. The first one, [email protected] (L.P.-C.) and [email protected] (V.L.) the hexokinase (HK), controls the conversion of glucose to glucose The online version of this article contains supplemental material. 6-phosphate. There are four isoenzymes of HK (HK-I, II, III, IV) Abbreviations used in this article: BM-DC, bone marrow–derived DC; DC, dendritic encoded by four different genes (HK1, 2, 3, 4). HK1 and HK3 have a cell; 2-DG, 2-deoxyglucose; Echino, echinomycin; HIF-1, hypoxia-inducible factor 1; HK, hexokinase; MoDC, monocyte-derived DC; mTORC1, mammalian target of rapa- large spectrum of tissue expression, whereas HK2 and HK4 have an mycin complex 1; 2-NBDG, 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D- expression profile that is more restricted to metabolically relevant glucose; Pam, Pam3CSK4; PFK, phosphofructokinase; PGN, peptidoglycan; PHD, tissues. HK2 is overexpressed in virtually all cancer cells. Other rate- prolyl-hydroxylase domain enzyme; SB, SB203580. limiting enzymes of glycolysis are the phosphofructokinase (PFK) Copyright Ó 2018 by The American Association of Immunologists, Inc. 0022-1767/18/$35.00 and pyruvate kinase. PFK is encoded by three different genes www.jimmunol.org/cgi/doi/10.4049/jimmunol.1701522 2 TLR4 GLYCOLYTIC BURST INVOLVES A p38–MAPK–HIF-1a AXIS (PFKL, PFKP, and PFKM) whose expression is tissue dependent. TLR1/2 ligand Pam3CSK4 (Pam), all from InvivoGen (Toulouse, France), Pyruvate kinase is encoded by the PKM gene in immune cells, were dissolved in sterile PBS. FP7 was synthesized from commercially generating two isoforms (PKM1 and PKM2). The pyruvate produced available D-glucose by multistep organic synthesis according to published procedures (32). The purity of the molecule was assessed by nuclear mag- by glycolysis can be converted to lactate that is excreted by specific netic resonance and mass spectrometry analysis. A 5-mM stock solution of monocarboxylate transporters or to oxaloacetate or acetyl-CoA to FP7 was prepared in ethanol/DMSO 1:1 and conserved at 220˚C. fuel the tricarboxylic acid cycle, coupling glycolysis to oxidative The inhibitors SB203580 (SB), SP600125, PD98059, BX795, and phosphorylation, for the generation of high amount of ATP in the LY294002, purchased from InvivoGen, rapamycin, provided by Calbiochem (San Diego, CA), and HIF-1a DNA-binding inhibitor, echinomycin (Echino), presence of oxygen. During hypoxia, oxidative phosphorylation is provided by Cayman Chemical (Ann Arbor, MI), were dissolved in sterile reduced and glycolysis is increased to face energetic needs (14). culture grade DMSO and stored at 220˚C. AZD5363 and SC75741 were However, activation of glycolysis can also occur under aerobic con- purchased from Selleckchem (Houston, TX), prepared in DMSO, and stored ditions, and O. Warburg (15) first discovered that tumor cells have a at 280˚C. Pepinh MYD, pepinh TRIF, and pepinh control, purchased from 2 high rate of glycolysis and that most pyruvate is converted to lactate, InvivoGen, were dissolved in PBS and stored at 20˚C.

View Full Text

Details

  • File Type
    pdf
  • Upload Time
    -
  • Content Languages
    English
  • Upload User
    Anonymous/Not logged-in
  • File Pages
    17 Page
  • File Size
    -

Download

Channel Download Status
Express Download Enable

Copyright

We respect the copyrights and intellectual property rights of all users. All uploaded documents are either original works of the uploader or authorized works of the rightful owners.

  • Not to be reproduced or distributed without explicit permission.
  • Not used for commercial purposes outside of approved use cases.
  • Not used to infringe on the rights of the original creators.
  • If you believe any content infringes your copyright, please contact us immediately.

Support

For help with questions, suggestions, or problems, please contact us