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Link to original content: https://doi.org/10.1038/s41591-018-0159-7
Metformin reduces liver glucose production by inhibition of fructose-1-6-bisphosphatase | Nature Medicine
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Metformin reduces liver glucose production by inhibition of fructose-1-6-bisphosphatase

Nature Medicine volume 24, pages 1395–1406 (2018)Cite this article

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Abstract

Metformin is a first-line drug for the treatment of individuals with type 2 diabetes, yet its precise mechanism of action remains unclear. Metformin exerts its antihyperglycemic action primarily through lowering hepatic glucose production (HGP). This suppression is thought to be mediated through inhibition of mitochondrial respiratory complex I, and thus elevation of 5′-adenosine monophosphate (AMP) levels and the activation of AMP-activated protein kinase (AMPK), though this proposition has been challenged given results in mice lacking hepatic AMPK. Here we report that the AMP-inhibited enzyme fructose-1,6-bisphosphatase-1 (FBP1), a rate-controlling enzyme in gluconeogenesis, functions as a major contributor to the therapeutic action of metformin. We identified a point mutation in FBP1 that renders it insensitive to AMP while sparing regulation by fructose-2,6-bisphosphate (F-2,6-P2), and knock-in (KI) of this mutant in mice significantly reduces their response to metformin treatment. We observe this during a metformin tolerance test and in a metformin-euglycemic clamp that we have developed. The antihyperglycemic effect of metformin in high-fat diet–fed diabetic FBP1-KI mice was also significantly blunted compared to wild-type controls. Collectively, we show a new mechanism of action for metformin and provide further evidence that molecular targeting of FBP1 can have antihyperglycemic effects.

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Fig. 1: Generation of the AMP-insensitive FBP1-G27P-KI mouse model.
Fig. 2: FBP1-G27P-KI mice display normal glucose homeostasis.
Fig. 3: FBP1-G27P-KI mice are resistant to the hypoglycemic action of an AMP-mimetic FBPase inhibitor.
Fig. 4: FBP1-G27P-KI mice are resistant to the hypoglycemic action of AICAR.
Fig. 5: FBP1-G27P-KI mice exhibit resistance to the acute glucose-lowering effect of metformin.
Fig. 6: FBP1-G27P-KI mice are resistant to the glucose lowering effects of metformin in an obesity-induced model of diabetes.

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Acknowledgements

We thank M. Deak for molecular biology assistance and S. Jakobsen and J. Frøkiær for support in method development of the [11C]metformin-uptake study. We also thank E. Heikkilä for performing islet isolation, S. Ducommun for performing the pTBC1D1 blot, and S. Cotting for constructing Wollenberger tongs.GLUT2 antibody was provided by B. Thorens, GCK/HXK4 antibody was provided by M. Magnuson, GCKR antibody was provided by M. Shiota, G6PC antibody was provided by G. Mithieux, PFKFB1 antibody was provided by S. Baltrusch, pS33 PFKFB1 antibody was provided by J. Xie and pS8 GYS2 antibody was provided by J. Guinovart. This study was supported by Vanderbilt Mouse Metabolic Phenotyping Center Grant DK059637 (D.H.W.) and R37 DK050277 (D.H.W.), a Foundation Grant (FND 143277) from the Canadian Institutes of Health Research (F.S.), the Danish Council for Independent Research DFF—4183-00384 (N.J.) and the Novo Nordisk Foundation NNF13OC0003882 (N.J.). E.Z. was supported by a Sir Henry Wellcome postdoctoral fellowship. C.C.H. was supported by a Canadian Diabetes Association postdoctoral fellowship.

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  1. Roger W. Hunter

    Present address: School of Physiology, Pharmacology & Neuroscience, University of Bristol, Bristol, UK

  2. Elton Zeqiraj

    Present address: Astbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK

Authors and Affiliations

  1. Nestlé Institute of Health Sciences SA, Lausanne, Switzerland

    Roger W. Hunter & Kei Sakamoto

  2. Department of Molecular Physiology and Biophysics and the Vanderbilt Mouse Metabolic Phenotyping Center, Vanderbilt University, Nashville, TN, USA

    Curtis C. Hughey, Louise Lantier & David H. Wasserman

  3. Departments of Clinical Medicine and Biomedicine, Aarhus University, Aarhus, Denmark

    Elias I. Sundelin & Niels Jessen

  4. MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee, UK

    Mark Peggie

  5. Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, ON, Canada

    Elton Zeqiraj & Frank Sicheri

  6. Departments of Biochemistry and Molecular Genetics, University of Toronto, Toronto, ON, Canada

    Frank Sicheri

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Contributions

R.W.H. and K.S. designed the study. R.W.H. performed all biochemical assays and the majority of in vivo experiments, assisted by K.S. Analysis of FBP1 structure and design of the mutants were performed by E.Z. and F.S. M.P. performed molecular cloning and mutagenesis of FBP1. N.J. and E.I.S. performed the [11C]metformin-uptake kinetics study and analyzed the data. C.C.H. and L.L. performed the metformin euglycemic clamp and analyzed the data. D.H.W. supervised C.C.H. and L.L. and contributed to interpretation of data from the clamp study. R.W.H and K.S. wrote the manuscript. All authors reviewed, edited and approved the manuscript.

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Correspondence to Kei Sakamoto.

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Hunter, R.W., Hughey, C.C., Lantier, L. et al. Metformin reduces liver glucose production by inhibition of fructose-1-6-bisphosphatase. Nat Med 24, 1395–1406 (2018). https://doi.org/10.1038/s41591-018-0159-7

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