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Slc12a8 is a nicotinamide mononucleotide transporter

An Author Correction to this article was published on 12 July 2019

Matters Arising to this article was published on 12 July 2019

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Abstract

Nicotinamide mononucleotide (NMN) is a biosynthetic precursor of nicotinamide adenine dinucleotide (NAD+) known to promote cellular NAD+ production and counteract age-associated pathologies associated with a decline in tissue NAD+ levels. How NMN is taken up into cells has not been entirely clear. Here we show that the Slc12a8 gene encodes a specific NMN transporter. We find that Slc12a8 is highly expressed and regulated by NAD+ in the mouse small intestine. Slc12a8 knockdown abrogates the uptake of NMN in vitro and in vivo. We further show that Slc12a8 specifically transports NMN, but not nicotinamide riboside, and that NMN transport depends on the presence of sodium ion. Slc12a8 deficiency significantly decreases NAD+ levels in the jejunum and ileum, which is associated with reduced NMN uptake as traced by doubly labelled isotopic NMN. Finally, we observe that Slc12a8 expression is upregulated in the aged mouse ileum, which contributes to the maintenance of ileal NAD+ levels. Our work identifies a specific NMN transporter and demonstrates that Slc12a8 has a critical role in regulating intestinal NAD+ metabolism.

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Fig. 1: Identification and characterization of the Slc12a8 gene.
Fig. 2: The kinetic features of the Slc12a8 NMN transporter and its specificity, sodium dependency and effects on NAD+ biosynthesis.
Fig. 3: In vivo knockdown of Slc12a8 in the small intestine.
Fig. 4: The age-associated upregulation of Slc12a8 in the ileum and its effect on NMN uptake and ileal NAD+ biosynthesis in aged mice.

Data availability

The microarray data used in this study has been deposited into the National Center for Biotechnology Information Gene Expression Omnibus (GEO) database (GEO accession numbers GSE49784 and GSE118365). All data generated or analysed during this study are included in the article and its Supplementary Information.

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  • 12 July 2019

    An amendment to this paper has been published and can be accessed via a link at the top of the paper.

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Acknowledgements

We thank C. Cantoni for flow cytometric analysis, L. Guida for support in proteoliposome preparation and R. Lewis for the production of Slc12a8-KO mice. We also thank E. Schulak for his generous support to A.G., members of the S.I. lab for critical comments and suggestions on this study and staff members in the core facilities provided by the Diabetes Research Center (P30 DK020579), the Nutrition Obesity Research Center (P30 DK56341) and the Hope Center for Neurological Disorders at Washington University. This work was also performed in a facility supported by NCRR grant C06 RR015502. A.G. was supported as the Tanaka Scholar by T. Tanaka and M. Tanaka. M.E.M was supported by UK Research Councils and Biotechnology and Biological Science Research Council (BBSRC; BB/N001842/1). This work was mainly supported by grants from the National Institute on Aging (AG024150, AG037457, AG047902) to S.I.

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Contributions

S.I. conceived the project, and A.G. and S.I. mainly designed research, analysed data and wrote the manuscript. A.G. performed most experiments. K.F.M. performed the analyses of gut-specific Slc12a8-KD mice and whole-body Slc12a8-KO mice with A.G. J.Y. performed microarray analyses. S.B. and G.S. conducted proteoliposome experiments and analysed the results with A.G. and S.I. K.T. and A.G. performed immunostaining, and H.C.L. and A.G. conducted Slc12a8 overexpression in vivo. R.C. and M.E.M. synthesized O18-D-NR and O18-D-NMN for this study, and Y.S. and A.G. conducted mass spectrometry analyses.

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Correspondence to Shin-ichiro Imai.

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A.G. and S.I. are inventors on a patent (PCT/US18/46233) about the Slc12a8 NMN transporter, whose applicant is Washington University and which has been licensed by Teijin Limited (Japan). Other authors declare no competing interests.

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Grozio, A., Mills, K.F., Yoshino, J. et al. Slc12a8 is a nicotinamide mononucleotide transporter. Nat Metab 1, 47–57 (2019). https://doi.org/10.1038/s42255-018-0009-4

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