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      Lactate Inhibits the Pro-Inflammatory Response and Metabolic Reprogramming in Murine Macrophages in a GPR81-Independent Manner

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          Abstract

          Lactate is an essential component of carbon metabolism in mammals. Recently, lactate was shown to signal through the G protein coupled receptor 81 (GPR81) and to thus modulate inflammatory processes. This study demonstrates that lactate inhibits pro-inflammatory signaling in a GPR81-independent fashion. While lipopolysaccharide (LPS) triggered expression of IL-6 and IL-12 p40, and CD40 in bone marrow-derived macrophages, lactate was able to abrogate these responses in a dose dependent manner in Gpr81 -/- cells as well as in wild type cells. Macrophage activation was impaired when glycolysis was blocked by chemical inhibitors. Remarkably, lactate was found to inhibit LPS-induced glycolysis in wild type as well as in Gpr81 -/- cells. In conclusion, our study suggests that lactate can induce GPR81-independent metabolic changes that modulate macrophage pro-inflammatory activation.

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          Most cited references19

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          Quantitative metabolome profiling of colon and stomach cancer microenvironment by capillary electrophoresis time-of-flight mass spectrometry.

          Most cancer cells predominantly produce energy by glycolysis rather than oxidative phosphorylation via the tricarboxylic acid (TCA) cycle, even in the presence of an adequate oxygen supply (Warburg effect). However, little has been reported regarding the direct measurements of global metabolites in clinical tumor tissues. Here, we applied capillary electrophoresis time-of-flight mass spectrometry, which enables comprehensive and quantitative analysis of charged metabolites, to simultaneously measure their levels in tumor and grossly normal tissues obtained from 16 colon and 12 stomach cancer patients. Quantification of 94 metabolites in colon and 95 metabolites in stomach involved in glycolysis, the pentose phosphate pathway, the TCA and urea cycles, and amino acid and nucleotide metabolisms resulted in the identification of several cancer-specific metabolic traits. Extremely low glucose and high lactate and glycolytic intermediate concentrations were found in both colon and stomach tumor tissues, which indicated enhanced glycolysis and thus confirmed the Warburg effect. Significant accumulation of all amino acids except glutamine in the tumors implied autophagic degradation of proteins and active glutamine breakdown for energy production, i.e., glutaminolysis. In addition, significant organ-specific differences were found in the levels of TCA cycle intermediates, which reflected the dependency of each tissue on aerobic respiration according to oxygen availability. The results uncovered unexpectedly poor nutritional conditions in the actual tumor microenvironment and showed that capillary electrophoresis coupled to mass spectrometry-based metabolomics, which is capable of quantifying the levels of energy metabolites in tissues, could be a powerful tool for the development of novel anticancer agents that target cancer-specific metabolism.
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            Tumor-derived lactic acid modulates dendritic cell activation and antigen expression.

            The tumor milieu can influence dendritic cell (DC) differentiation. We analyzed DC differentiation in a 3-dimensional tumor model and propose a new mechanism of DC modulation by the tumor environment. Monocytes were cultured in the presence of IL-4 and GM-CSF within multicellular tumor spheroids (MCTSs) generated from different tumor cell lines. Monocytes invaded the MCTSs and differentiated into tumor-associated dendritic cells (TADCs). The antigen expression was altered on TADCs independent of the culture conditions (immature/mature DCs, Langerhans cells) and IL-12 secretion was reduced. Supernatants of MCTSs could partially transfer the suppressive effect. Conditioned media from urothelial carcinoma cell lines contained high levels of M-CSF and IL-6, both cytokines known to modulate DC differentiation. In contrast, melanoma and prostate carcinoma MCTS cocultures produced little M-CSF and IL-6, but high levels of lactic acid. Indeed, addition of lactic acid during DC differentiation in vitro induced a phenotype comparable with TADCs generated within melanoma and prostate carcinoma MCTSs. Blocking of lactic acid production in melanoma MCTS cocultures reverted the TADC phenotype to normal. We therefore conclude that tumor-derived lactic acid is an important factor modulating the DC phenotype in the tumor environment, which may critically contribute to tumor escape mechanisms.
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              Lactate reduces liver and pancreatic injury in Toll-like receptor- and inflammasome-mediated inflammation via GPR81-mediated suppression of innate immunity.

              The NACHT, LRR, and pyrin domain-containing protein 3 (NLRP3) inflammasome induces inflammation in response to organ injury, but little is known about its regulation. Toll-like receptors (TLRs) provide the first signal required for activation of the inflammasome and stimulate aerobic glycolysis to generate lactate. We examined whether lactate and the lactate receptor, Gi-protein-coupled receptor 81 (GPR81), regulate TLR induction of signal 1 and limit inflammasome activation and organ injury.

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                15 November 2016
                2016
                : 11
                : 11
                : e0163694
                Affiliations
                [1 ]Instituto de Estudios Inmunológicos y Fisiopatológicos–CONICET- Nacional Universtity of La Plata, 1900, La Plata, Argentina
                [2 ]Institut Pasteur de Lille, Inserm, CNRS, Univ. Lille, CHU Lille, U1019—UMR8204—CIIL—Center for Infection and Immunity of Lille, F-59000, Lille, France
                [3 ]Univ. Lille, Inserm, CHU Lille, UMR-S 1172—JPArc—Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer, F-59000, Lille, France
                [4 ]Department of Pharmacology, Max Planck Institute for Heart and Lung Research, D-61231, Bad Nauheim, Germany
                [5 ]Medical Faculty, J.W. Goethe University, D-60590, Frankfurt, Germany
                Institut National de la Santeet de la Recherche Medicale (INSERM), FRANCE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: AE SO JCS MR.

                • Formal analysis: AE JCS MR.

                • Funding acquisition: SO JCS MR.

                • Investigation: AE JCS MR.

                • Methodology: AE DC PM CT JK.

                • Project administration: JCS MR.

                • Resources: SO JCS MR.

                • Supervision: JCS MR.

                • Visualization: AE SO JCS MR.

                • Writing – original draft: AE MR.

                • Writing – review & editing: JCS MR.

                Article
                PONE-D-16-19632
                10.1371/journal.pone.0163694
                5112849
                27846210
                a6324281-2bac-4460-8b0a-726c33f157bf
                © 2016 Errea et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 17 May 2016
                : 13 September 2016
                Page count
                Figures: 5, Tables: 0, Pages: 11
                Funding
                The work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica (ANPCYT), INSERM, CNRS, Institut Pasteur de Lille, Université de Lille, CONICET-DAAD and the Argentinian Ministry of Science, Technology and Innovation and the French Ministry for Research and Higher Education (grant ECOS A12B03). AE received a Bernardo Houssay fellowship from CONICET. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cellular Types
                Animal Cells
                Blood Cells
                White Blood Cells
                Macrophages
                Biology and Life Sciences
                Cell Biology
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                Animal Cells
                Immune Cells
                White Blood Cells
                Macrophages
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                Immunology
                Immune Cells
                White Blood Cells
                Macrophages
                Medicine and Health Sciences
                Immunology
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                White Blood Cells
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                Biochemistry
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