Systems-level metabolic flux profiling identifies fatty acid synthesis as a target for antiviral therapy. Nat Biotechnol

Department of Molecular Biology, Lewis Thomas Laboratory, Princeton University, Princeton, New Jersey 08544, USA.
Nature Biotechnology (Impact Factor: 41.51). 10/2008; 26(10):1179-86. DOI: 10.1038/nbt.1500
Source: PubMed


Viruses rely on the metabolic network of their cellular hosts to provide energy and building blocks for viral replication. We developed a flux measurement approach based on liquid chromatography-tandem mass spectrometry to quantify changes in metabolic activity induced by human cytomegalovirus (HCMV). This approach reliably elucidated fluxes in cultured mammalian cells by monitoring metabolome labeling kinetics after feeding cells (13)C-labeled forms of glucose and glutamine. Infection with HCMV markedly upregulated flux through much of the central carbon metabolism, including glycolysis. Particularly notable increases occurred in flux through the tricarboxylic acid cycle and its efflux to the fatty acid biosynthesis pathway. Pharmacological inhibition of fatty acid biosynthesis suppressed the replication of both HCMV and influenza A, another enveloped virus. These results show that fatty acid synthesis is essential for the replication of two divergent enveloped viruses and that systems-level metabolic flux profiling can identify metabolic targets for antiviral therapy.


Available from: Thomas Shenk, Oct 24, 2014
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    • "The fatty acids and other lipid materials that are generated by these processes are essential substrates for energy metabolism (Menendez and Lupu, 2007; Mashima et al., 2009) as well as the major components of all biological lipid membranes (Menendez and Lupu, 2007; Vander Heiden et al., 2009; Mashima et al., 2009). The Warburg Effect is also seen in vertebrate cells infected by oncogenic viruses such as KSHV (Kaposi's sarcoma-associated herpesvirus) (Delgado et al., 2010) and HCMV (Human cytomegalovirus ) (Munger et al., 2008), and a recent paper (Chen et al., 2011) further showed that it occurs in shrimp hemocytes that are infected by the White Spot Syndrome Virus (WSSV). WSSV, which is a large enveloped (approximately 300 kbp) dsDNA invertebrate virus with an in vivo replication cycle of 22e24 h, was found to trigger the Warburg effect at the WSSV genome replication stage (12 hpi [hours post infection]) via the activation of the PI3K-Akt-mTOR pathway (Su et al., 2014). "
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    ABSTRACT: White spot syndrome virus (WSSV), the causative agent of white spot disease (WSD), is a serious and aggressive shrimp viral pathogen with a worldwide distribution. At the genome replication stage (12 hpi), WSSV induces a metabolic rerouting known as the invertebrate Warburg effect, which boosts the availability of energy and biosynthetic building blocks in the host cell. Here we show that unlike the lipogenesis that is seen in cancer cells that are undergoing the Warburg effect, at 12 hpi, all of the long chain fatty acids (LCFAs) were significantly decreased in the stomach cells of WSSV-infected shrimp. By means of this non-selective WSSV-induced lipolysis, the LCFAs were apparently diverted into β-oxidation and used to replenish the TCA cycle. Conversely, at 24 hpi, when the Warburg effect had ceased, most of the LCFAs were significantly up-regulated and the composition was also significantly altered. In crayfish these changes were in a direction that appeared to favor the formation of WSSV virion particles. We also found that, at 24 hpi, but not at 12 hpi, the PI3K-Akt-mTOR-HIF1α pathway induced the expression of fatty acid synthase (FAS), an enzyme which catalyzes the conversion of acetyl-CoA into LCFAs. WSSV virion formation was impaired in the presence of the FAS inhibitor C75, although viral gene and viral DNA levels were unaffected. WSSV therefore appears to use the PI3K-Akt-mTOR pathway to induce lipid biosynthesis at 24 hpi in order to support viral morphogenesis. Copyright © 2015. Published by Elsevier Ltd.
    Developmental and comparative immunology 06/2015; 53(1). DOI:10.1016/j.dci.2015.06.001 · 2.82 Impact Factor
    • "ethanol (70 C). Supernatant of extracted samples were dried under vacuum and resuspended in LC-MS grade water for analysis of the relative abundance of 13 C and 15 N metabolites (Munger et al., 2008). "
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    ABSTRACT: Macrophage polarization involves a coordinated metabolic and transcriptional rewiring that is only partially understood. By using an integrated high-throughput transcriptional-metabolic profiling and analysis pipeline, we characterized systemic changes during murine macrophage M1 and M2 polarization. M2 polarization was found to activate glutamine catabolism and UDP-GlcNAc-associated modules. Correspondingly, glutamine deprivation or inhibition of N-glycosylation decreased M2 polarization and production of chemokine CCL22. In M1 macrophages, we identified a metabolic break at Idh, the enzyme that converts isocitrate to alpha-ketoglutarate, providing mechanistic explanation for TCA cycle fragmentation. (13)C-tracer studies suggested the presence of an active variant of the aspartate-arginosuccinate shunt that compensated for this break. Consistently, inhibition of aspartate-aminotransferase, a key enzyme of the shunt, inhibited nitric oxide and interleukin-6 production in M1 macrophages, while promoting mitochondrial respiration. This systems approach provides a highly integrated picture of the physiological modules supporting macrophage polarization, identifying potential pharmacologic control points for both macrophage phenotypes. Copyright © 2015 Elsevier Inc. All rights reserved.
    Immunity 03/2015; 42(3):419-30. DOI:10.1016/j.immuni.2015.02.005 · 21.56 Impact Factor
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    • "MS data were analyzed using MAVEN: Metabolomic Analysis and Visualization Engine (Clasquin et al., 2012; Melamud et al., 2010) and MATLAB (The MathWorks). For labeling experiments, the results were corrected for naturally occurring carbon-13 (Munger et al., 2008). "
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    ABSTRACT: Human cytomegalovirus (HCMV) infection rewires host-cell metabolism, upregulating flux from glucose into acetyl-CoA to feed fatty acid metabolism, with saturated very-long-chain fatty acids (VLFCAs) required for production of infectious virion progeny. The human genome encodes seven elongase enzymes (ELOVL) that extend long-chain fatty acids into VLCFA. Here, we identify ELOVL7 as pivotal for HCMV infection. HCMV induces ELOVL7 by more than 150-fold. This induction is dependent on mTOR and SREBP-1. ELOVL7 knockdown or mTOR inhibition impairs HCMV-induced fatty acid elongation, HCMV particle release, and infectivity per particle. ELOVL7 overexpression enhances HCMV replication. During HCMV infection, mTOR activity is maintained by the viral protein pUL38. Expression of pUL38 is sufficient to induce ELOVL7, and pUL38-deficient virus is partially defective in ELOVL7 induction and fatty acid elongation. Thus, through its ability to modulate mTOR and SREBP-1, HCMV induces ELOVL7 to synthesize the saturated VLCFA required for efficient virus replication. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
    Cell Reports 03/2015; 10(8-8):1375-1385. DOI:10.1016/j.celrep.2015.02.003 · 8.36 Impact Factor
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