Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer

Victor H Villar; Maria Francesca Allega; Ruhi Deshmukh; Tobias Ackermann; Mark A. Nakasone; Johan Vande Voorde; Thomas Drake; Janina  Oetjen; Algernon Bloom; Colin Nixon; Lars Vereecke; Maude  Jans; Gillian Blancke; Thomas G Bird; David  Lewis; Owen J Sansom; Karen Blyth; David  Sumpton; Saverio Tardito

doi:10.1038/s41589-022-01154-9

Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer

Victor H Villar, Maria Francesca Allega, Ruhi Deshmukh, Tobias Ackermann, Mark A. Nakasone, Johan Vande Voorde, Thomas Drake, Janina Oetjen, Algernon Bloom, Colin Nixon, Lars Vereecke, Maude Jans, Gillian Blancke, Thomas G Bird, David Lewis, Owen J Sansom, Karen Blyth, David Sumpton, Saverio Tardito^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Glutamine synthetase (GS) is the only known enzyme that synthesizes glutamine in mammals. Here we show that mammalian GS utilizes glutamate and methylamine to produce a methylated glutamine analogue, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific deletion of Glul and pharmacologic inhibition of GS, suppress the hepatic and circulating levels of N5-methylglutamine in mice. Conversely, N5-methylglutamine levels are sustained by the metabolism of gut microbiome. This alternative activity of GS was confirmed using human recombinant enzyme and cells where a pathogenic mutation promotes the synthesis of N5-methylglutamine over glutamine. Finally, we show that the urine level of N5-methylglutamine correlates with tumour burden in a novel -catenin-driven model of liver cancer with high GS expression, disclosing the potential of N5-methylglutamine as a biomarker for GS-positive hepatocellular carcinoma.

Original language	English
Journal	Nature Chemical Biology
Early online date	24 Oct 2022
DOIs	https://doi.org/10.1038/s41589-022-01154-9
Publication status	E-pub ahead of print - 24 Oct 2022

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Villar, V. H., Allega, M. F., Deshmukh, R., Ackermann, T., Nakasone, M. A., Voorde, J. V., Drake, T., Oetjen, J., Bloom, A., Nixon, C., Vereecke, L., Jans, M., Blancke, G., Bird, T. G., Lewis, D., Sansom, O. J., Blyth, K., Sumpton, D., & Tardito, S. (2022). Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer. Nature Chemical Biology. Advance online publication. https://doi.org/10.1038/s41589-022-01154-9

@article{755800d752df444badf5ca0d7747c97d,

title = "Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer",

abstract = "Glutamine synthetase (GS) is the only known enzyme that synthesizes glutamine in mammals. Here we show that mammalian GS utilizes glutamate and methylamine to produce a methylated glutamine analogue, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific deletion of Glul and pharmacologic inhibition of GS, suppress the hepatic and circulating levels of N5-methylglutamine in mice. Conversely, N5-methylglutamine levels are sustained by the metabolism of gut microbiome. This alternative activity of GS was confirmed using human recombinant enzyme and cells where a pathogenic mutation promotes the synthesis of N5-methylglutamine over glutamine. Finally, we show that the urine level of N5-methylglutamine correlates with tumour burden in a novel -catenin-driven model of liver cancer with high GS expression, disclosing the potential of N5-methylglutamine as a biomarker for GS-positive hepatocellular carcinoma.",

author = "Villar, {Victor H} and Allega, {Maria Francesca} and Ruhi Deshmukh and Tobias Ackermann and Nakasone, {Mark A.} and Voorde, {Johan Vande} and Thomas Drake and Janina Oetjen and Algernon Bloom and Colin Nixon and Lars Vereecke and Maude Jans and Gillian Blancke and Bird, {Thomas G} and David Lewis and Sansom, {Owen J} and Karen Blyth and David Sumpton and Saverio Tardito",

note = "Funding Information: We thank the Core Services and Advanced Technologies at the Cancer Research UK Beatson Institute (C596/A17196 and A31287) and particularly the Metabolomics, Translational Molecular Imaging, Biological Services Unit, Histology Service and Molecular Technologies. We acknowledge M. Mazzone (KU Leuven) for providing the mice carrying the Glul allele, U. Kutay (ETH Zurich) for providing the GLUL pcDNA/FRT/TOcHA (wt, R324A, R324C) plasmids, J.P. Morton for providing the KPC mice and S. Niclou (Luxembourg Institute of Health) for providing the T16 glioblastoma cells. We acknowledge C. Winchester for comments on the manuscript and all members of the Oncometabolism laboratory and the Metabolomics Facility for constructive discussions. This work was funded by Cancer Research UK awards A17196 and A31287 (CRUK Beatson Institute), Cancer Research UK RadNet Glasgow Centre award A28803 (A.B.), Wellcome Trust grant WT107492Z (T.G.B. and M.M.), Cancer Research UK HUNTER accelerator award A26813 (E.H.T.), Cancer Research UK award A25045 and DRCQQR-May21\100002 (O.J.S.), Cancer Research UK award A29256 (D.T.H.), Cancer Research UK award A29799 (K.B.), Cancer Research UK award A25006 (D.Y.L.) and Cancer Research UK award A23982 (S.T.). tm3Whla fl Publisher Copyright: {\textcopyright} 2022, The Author(s).",

year = "2022",

month = oct,

day = "24",

doi = "10.1038/s41589-022-01154-9",

language = "English",

journal = "Nature Chemical Biology",

issn = "1552-4450",

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Villar, VH, Allega, MF, Deshmukh, R, Ackermann, T, Nakasone, MA, Voorde, JV, Drake, T, Oetjen, J, Bloom, A, Nixon, C, Vereecke, L, Jans, M, Blancke, G, Bird, TG, Lewis, D, Sansom, OJ, Blyth, K, Sumpton, D & Tardito, S 2022, 'Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer', Nature Chemical Biology. https://doi.org/10.1038/s41589-022-01154-9

TY - JOUR

T1 - Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer

AU - Villar, Victor H

AU - Allega, Maria Francesca

AU - Deshmukh, Ruhi

AU - Ackermann, Tobias

AU - Nakasone, Mark A.

AU - Voorde, Johan Vande

AU - Drake, Thomas

AU - Oetjen, Janina

AU - Bloom, Algernon

AU - Nixon, Colin

AU - Vereecke, Lars

AU - Jans, Maude

AU - Blancke, Gillian

AU - Bird, Thomas G

AU - Lewis, David

AU - Sansom, Owen J

AU - Blyth, Karen

AU - Sumpton, David

AU - Tardito, Saverio

N1 - Funding Information: We thank the Core Services and Advanced Technologies at the Cancer Research UK Beatson Institute (C596/A17196 and A31287) and particularly the Metabolomics, Translational Molecular Imaging, Biological Services Unit, Histology Service and Molecular Technologies. We acknowledge M. Mazzone (KU Leuven) for providing the mice carrying the Glul allele, U. Kutay (ETH Zurich) for providing the GLUL pcDNA/FRT/TOcHA (wt, R324A, R324C) plasmids, J.P. Morton for providing the KPC mice and S. Niclou (Luxembourg Institute of Health) for providing the T16 glioblastoma cells. We acknowledge C. Winchester for comments on the manuscript and all members of the Oncometabolism laboratory and the Metabolomics Facility for constructive discussions. This work was funded by Cancer Research UK awards A17196 and A31287 (CRUK Beatson Institute), Cancer Research UK RadNet Glasgow Centre award A28803 (A.B.), Wellcome Trust grant WT107492Z (T.G.B. and M.M.), Cancer Research UK HUNTER accelerator award A26813 (E.H.T.), Cancer Research UK award A25045 and DRCQQR-May21\100002 (O.J.S.), Cancer Research UK award A29256 (D.T.H.), Cancer Research UK award A29799 (K.B.), Cancer Research UK award A25006 (D.Y.L.) and Cancer Research UK award A23982 (S.T.). tm3Whla fl Publisher Copyright: © 2022, The Author(s).

PY - 2022/10/24

Y1 - 2022/10/24

N2 - Glutamine synthetase (GS) is the only known enzyme that synthesizes glutamine in mammals. Here we show that mammalian GS utilizes glutamate and methylamine to produce a methylated glutamine analogue, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific deletion of Glul and pharmacologic inhibition of GS, suppress the hepatic and circulating levels of N5-methylglutamine in mice. Conversely, N5-methylglutamine levels are sustained by the metabolism of gut microbiome. This alternative activity of GS was confirmed using human recombinant enzyme and cells where a pathogenic mutation promotes the synthesis of N5-methylglutamine over glutamine. Finally, we show that the urine level of N5-methylglutamine correlates with tumour burden in a novel -catenin-driven model of liver cancer with high GS expression, disclosing the potential of N5-methylglutamine as a biomarker for GS-positive hepatocellular carcinoma.

AB - Glutamine synthetase (GS) is the only known enzyme that synthesizes glutamine in mammals. Here we show that mammalian GS utilizes glutamate and methylamine to produce a methylated glutamine analogue, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific deletion of Glul and pharmacologic inhibition of GS, suppress the hepatic and circulating levels of N5-methylglutamine in mice. Conversely, N5-methylglutamine levels are sustained by the metabolism of gut microbiome. This alternative activity of GS was confirmed using human recombinant enzyme and cells where a pathogenic mutation promotes the synthesis of N5-methylglutamine over glutamine. Finally, we show that the urine level of N5-methylglutamine correlates with tumour burden in a novel -catenin-driven model of liver cancer with high GS expression, disclosing the potential of N5-methylglutamine as a biomarker for GS-positive hepatocellular carcinoma.

U2 - 10.1038/s41589-022-01154-9

DO - 10.1038/s41589-022-01154-9

M3 - Article

SN - 1552-4450

JO - Nature Chemical Biology

JF - Nature Chemical Biology

ER -

Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer

Abstract

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