Abstract

In obesity and type 2 diabetes, Glut4 glucose transporter expression is decreased selectively in adipocytes1. Adipose-specific knockout or overexpression of Glut4 alters systemic insulin sensitivity2. Here we show, using DNA array analyses, that nicotinamide N-methyltransferase (Nnmt) is the most strongly reciprocally regulated gene when comparing gene expression in white adipose tissue (WAT) from adipose-specific Glut4-knockout or adipose-specific Glut4-overexpressing mice with their respective controls. NNMT methylates nicotinamide (vitamin B3) using S-adenosylmethionine (SAM) as a methyl donor3,4. Nicotinamide is a precursor of NAD+, an important cofactor linking cellular redox states with energy metabolism5. SAM provides propylamine for polyamine biosynthesis and donates a methyl group for histone methylation6. Polyamine flux including synthesis, catabolism and excretion, is controlled by the rate-limiting enzymes ornithine decarboxylase (ODC) and spermidine–spermine N1-acetyltransferase (SSAT; encoded by Sat1) and by polyamine oxidase (PAO), and has a major role in energy metabolism7,8. We report that NNMT expression is increased in WAT and liver of obese and diabetic mice. Nnmt knockdown in WAT and liver protects against diet-induced obesity by augmenting cellular energy expenditure. NNMT inhibition increases adipose SAM and NAD+ levels and upregulates ODC and SSAT activity as well as expression, owing to the effects of NNMT on histone H3 lysine 4 methylation in adipose tissue. Direct evidence for increased polyamine flux resulting from NNMT inhibition includes elevated urinary excretion and adipocyte secretion of diacetylspermine, a product of polyamine metabolism. NNMT inhibition in adipocytes increases oxygen consumption in an ODC-, SSAT- and PAO-dependent manner. Thus, NNMT is a novel regulator of histone methylation, polyamine flux and NAD+-dependent SIRT1 signalling, and is a unique and attractive target for treating obesity and type 2 diabetes.

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Accessions

Primary accessions

Gene Expression Omnibus

Data deposits

The microarray data from adipose tissue of adipose-specific knockout and adipose-specific overexpression have been published by our laboratory and are available in NCBI Gene Expression Omnibus under accession number GSE35378.

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Acknowledgements

We thank R. Weinshilboum for NNMT antibody; P. Woster for DFMO; M. Yuan for tandem mass spectrometry; A. Karppinen, A. Korhonen, T. Reponen, A. Uimari, S. Pirnes-Karhu and T. Koponen for measurements of ODC and SSAT activity; C. Semenkovich and S. Fried for protocols for FAS activity measurements; and P. Aryal for assistance with real-time qPCR. D.Kr. is supported by the Deutsche Forschungsgemeinschaft (KR 3475/1-1) and American Heart Association (AHA) (09POST2250499); Q.Y. is a Klarman Scholar at the Beth Israel Deaconess Medical Center. This work is supported by grants from the NIH (R37 DK43051, P30 DK57521) and a grant from the JPB foundation to B.B.K.; grants from the NIH (KO8 DK090149, R01 DK100385, BNORC P30 DK046200 and NORCH P30 DK040561) to Q.Y.; grant RO1 DK69966 to P.P.; P01CA120964 and P30CA006516-46 to J.M.A.; AHA 13SDG14620005 and P&F P30 DK0460200 to D.K.; the Ellison Medical Foundation New Scholar in Aging Award to A.A.S.; and academy of Finland grant 118590 to L.A.

Author information

Author notes

    • Daniel Kraus
    •  & Qin Yang

    These authors contributed equally to this work.

    • Daniel Kraus
    • , Qin Yang
    • , Eija Pirinen
    • , Thomas C. Pulinilkunnil
    • , Fengying Gong
    •  & Leena Alhonen

    Present addresses: Division of Nephrology, Department of Internal Medicine I, Würzburg University Hospital, Oberdürrbacher Straße 6, 97080 Würzburg, Germany (D.K.); Department of Medicine, Physiology and Biophysics, Center for Diabetes Research and Treatment, and Center for Epigenetics and Metabolism, University of California, Irvine, California 92697, USA (Q.Y.); Research Programs Unit, Molecular Neurology, Biomedicum Helsinki, University of Helsinki, 00290, Helsinki, Finland (E.P.); Department of Biochemistry and Molecular Biology, Faculty of Medicine, Dalhousie Medicine New Brunswick, Dalhousie University, Saint John, New Brunswick E2L4L5, USA (T.C.P.); Department of Endocrinology, Key Laboratory of Endocrinology of Ministry of Health, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China (F.G.); School of Pharmacy, University of Eastern Finland, P.O. Box 1627, FI-70211 Kuopio, Finland (L.A.).

Affiliations

  1. Division of Endocrinology, Diabetes, and Metabolism, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, 330 Brookline Avenue, Boston, Massachusetts 02215, USA

    • Daniel Kraus
    • , Qin Yang
    • , Dong Kong
    • , Lin Zhang
    • , Thomas C. Pulinilkunnil
    • , Fengying Gong
    • , Ya-chin Wang
    • , Odile D. Peroni
    •  & Barbara B. Kahn
  2. Department of Cancer Biology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Alexander S. Banks
    • , Joseph T. Rodgers
    •  & Pere Puigserver
  3. Biotechnology and Molecular Medicine, A.I. Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Eastern Finland, Kuopio Campus, PO Box 1627, FI-70211 Kuopio, Finland

    • Eija Pirinen
    •  & Leena Alhonen
  4. Department of Pharmacology, Weill Medical College of Cornell University, 1300 York Avenue, New York, New York 10065, USA

    • Yana Cen
    •  & Anthony A. Sauve
  5. Division of Signal Transduction, Beth Israel Deaconess Medical Center and Harvard Medical School, 330 Brookline Ave, Boston, Massachusetts 02215, USA

    • John M. Asara
  6. Isis Pharmaceuticals, 1896 Rutherford Road, Carlsbad, California 92008-7326, USA

    • Brett P. Monia
    •  & Sanjay Bhanot

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Contributions

Q.Y. discovered NNMT from the initial microarray analysis. D.Kr., Q.Y. and B.B.K designed the experiments, interpreted the data and wrote the paper. D.Ko. performed oxygen consumption experiments in adipocytes. A.S.B. performed CLAMS studies. L.Z., T.C.P., F.G., YC.W. and O.D.P. provided assistance with cell culture and animal experiments. O.D.P. also performed the microarray studies. J.T.R. and P.P. performed PGC-1α acetylation experiments. E.P. and L.A. provided expertise on polyamines and measured ODC and SSAT activity. Y.C. and A.A.S. measured nicotinamide and metabolites. J.M.A. performed metabolomics studies. B.P.M and S.B. provided Nnmt and control ASOs.

Competing interests

B.B.K., Q.Y. and D. Kr. are inventors on a patent application related to NNMT. B.P.M. and S.B. are employees of Isis Pharmaceuticals Inc.

Corresponding authors

Correspondence to Qin Yang or Barbara B. Kahn.

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DOI

https://doi.org/10.1038/nature13198

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