Hepatic glutamine synthetase controls N5-methylglutamine in homeostasis and cancer

VH Villar; MF Allega; R Deshmukh; T Ackermann; MA Nakasone; J Vande Voorde; TM Drake; J Oetjen; A Bloom; C Nixon; M Müller; S May; EH Tan; L Vereecke; M Jans; G Blancke; DJ Murphy; DT Huang; DY Lewis; TG Bird; OJ Sansom; K Blyth; D Sumpton; S Tardito

doi:10.1038/s41589-022-01154-9

Hepatic glutamine synthetase controls N⁵-methylglutamine in homeostasis and cancer

VH Villar, MF Allega, R Deshmukh, T Ackermann, MA Nakasone, J Vande Voorde, TM Drake, J Oetjen, A Bloom, C Nixon, M Müller, S May, EH Tan, L Vereecke, M Jans, G Blancke, DJ Murphy, DT Huang, DY Lewis, TG BirdOJ Sansom, K Blyth, D Sumpton, S Tardito^*

^*Corresponding author for this work

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

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Abstract

Glutamine synthetase (GS) activity is conserved from prokaryotes to humans, where the ATP-dependent production of glutamine from glutamate and ammonia is essential for neurotransmission and ammonia detoxification. Here, we show that mammalian GS uses glutamate and methylamine to produce a methylated glutamine analog, N⁵-methylglutamine. Untargeted metabolomics revealed that liver-specific GS deletion and its pharmacological inhibition in mice suppress hepatic and circulating levels of N⁵-methylglutamine. This alternative activity of GS was confirmed in human recombinant enzyme and cells, where a pathogenic mutation in the active site (R324C) promoted the synthesis of N⁵-methylglutamine over glutamine. N⁵-methylglutamine is detected in the circulation, and its levels are sustained by the microbiome, as demonstrated by using germ-free mice. Finally, we show that urine levels of N⁵-methylglutamine correlate with tumor burden and GS expression in a β-catenin-driven model of liver cancer, highlighting the translational potential of this uncharacterized metabolite.

Original language	English
Pages (from-to)	292-304
Number of pages	13
Journal	Nature Chemical Biology
Volume	19
Issue number	3
Early online date	24 Oct 2022
DOIs	https://doi.org/10.1038/s41589-022-01154-9
Publication status	Published - Mar 2023

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Cite this

Villar, VH., Allega, MF., Deshmukh, R., Ackermann, T., Nakasone, MA., Vande Voorde, J., Drake, TM., Oetjen, J., Bloom, A., Nixon, C., Müller, M., May, S., Tan, EH., Vereecke, L., Jans, M., Blancke, G., Murphy, DJ., Huang, DT., Lewis, DY., ... Tardito, S. (2023). Hepatic glutamine synthetase controls N⁵-methylglutamine in homeostasis and cancer. Nature Chemical Biology, 19(3), 292-304. https://doi.org/10.1038/s41589-022-01154-9

@article{290938fdf14e4159b6090ec99f055404,

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

abstract = "Glutamine synthetase (GS) activity is conserved from prokaryotes to humans, where the ATP-dependent production of glutamine from glutamate and ammonia is essential for neurotransmission and ammonia detoxification. Here, we show that mammalian GS uses glutamate and methylamine to produce a methylated glutamine analog, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific GS deletion and its pharmacological inhibition in mice suppress hepatic and circulating levels of N5-methylglutamine. This alternative activity of GS was confirmed in human recombinant enzyme and cells, where a pathogenic mutation in the active site (R324C) promoted the synthesis of N5-methylglutamine over glutamine. N5-methylglutamine is detected in the circulation, and its levels are sustained by the microbiome, as demonstrated by using germ-free mice. Finally, we show that urine levels of N5-methylglutamine correlate with tumor burden and GS expression in a β-catenin-driven model of liver cancer, highlighting the translational potential of this uncharacterized metabolite.",

author = "VH Villar and MF Allega and R Deshmukh and T Ackermann and MA Nakasone and {Vande Voorde}, J and TM Drake and J Oetjen and A Bloom and C Nixon and M M{\"u}ller and S May and EH Tan and L Vereecke and M Jans and G Blancke and DJ Murphy and DT Huang and DY Lewis and TG Bird and OJ Sansom and K Blyth and D Sumpton and S Tardito",

note = "Funding: 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.).",

year = "2023",

month = mar,

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

language = "English",

volume = "19",

pages = "292--304",

journal = "Nature Chemical Biology",

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Villar, VH, Allega, MF, Deshmukh, R, Ackermann, T, Nakasone, MA, Vande Voorde, J, Drake, TM, Oetjen, J, Bloom, A, Nixon, C, Müller, M, May, S, Tan, EH, Vereecke, L, Jans, M, Blancke, G, Murphy, DJ, Huang, DT, Lewis, DY, Bird, TG, Sansom, OJ, Blyth, K, Sumpton, D & Tardito, S 2023, 'Hepatic glutamine synthetase controls N⁵-methylglutamine in homeostasis and cancer', Nature Chemical Biology, vol. 19, no. 3, pp. 292-304. https://doi.org/10.1038/s41589-022-01154-9

TY - JOUR

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

AU - Villar, VH

AU - Allega, MF

AU - Deshmukh, R

AU - Ackermann, T

AU - Nakasone, MA

AU - Vande Voorde, J

AU - Drake, TM

AU - Oetjen, J

AU - Bloom, A

AU - Nixon, C

AU - Müller, M

AU - May, S

AU - Tan, EH

AU - Vereecke, L

AU - Jans, M

AU - Blancke, G

AU - Murphy, DJ

AU - Huang, DT

AU - Lewis, DY

AU - Bird, TG

AU - Sansom, OJ

AU - Blyth, K

AU - Sumpton, D

AU - Tardito, S

N1 - Funding: 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.).

PY - 2023/3

Y1 - 2023/3

N2 - Glutamine synthetase (GS) activity is conserved from prokaryotes to humans, where the ATP-dependent production of glutamine from glutamate and ammonia is essential for neurotransmission and ammonia detoxification. Here, we show that mammalian GS uses glutamate and methylamine to produce a methylated glutamine analog, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific GS deletion and its pharmacological inhibition in mice suppress hepatic and circulating levels of N5-methylglutamine. This alternative activity of GS was confirmed in human recombinant enzyme and cells, where a pathogenic mutation in the active site (R324C) promoted the synthesis of N5-methylglutamine over glutamine. N5-methylglutamine is detected in the circulation, and its levels are sustained by the microbiome, as demonstrated by using germ-free mice. Finally, we show that urine levels of N5-methylglutamine correlate with tumor burden and GS expression in a β-catenin-driven model of liver cancer, highlighting the translational potential of this uncharacterized metabolite.

AB - Glutamine synthetase (GS) activity is conserved from prokaryotes to humans, where the ATP-dependent production of glutamine from glutamate and ammonia is essential for neurotransmission and ammonia detoxification. Here, we show that mammalian GS uses glutamate and methylamine to produce a methylated glutamine analog, N5-methylglutamine. Untargeted metabolomics revealed that liver-specific GS deletion and its pharmacological inhibition in mice suppress hepatic and circulating levels of N5-methylglutamine. This alternative activity of GS was confirmed in human recombinant enzyme and cells, where a pathogenic mutation in the active site (R324C) promoted the synthesis of N5-methylglutamine over glutamine. N5-methylglutamine is detected in the circulation, and its levels are sustained by the microbiome, as demonstrated by using germ-free mice. Finally, we show that urine levels of N5-methylglutamine correlate with tumor burden and GS expression in a β-catenin-driven model of liver cancer, highlighting the translational potential of this uncharacterized metabolite.

U2 - 10.1038/s41589-022-01154-9

DO - 10.1038/s41589-022-01154-9

M3 - Article

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SN - 1552-4450

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JO - Nature Chemical Biology

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