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Characterization of the melanocortin-4-receptor nonsense mutation W16X in vitro and in vivo

The Pharmacogenomics Journal volume 13, pages 80–93 (2013)Cite this article

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Abstract

Several genetic diseases are triggered by nonsense mutations leading to the formation of truncated and defective proteins. Aminoglycosides have the capability to mediate a bypass of stop mutations during translation thus resulting in a rescue of protein expression. So far no attention has been directed to obesity-associated stop mutations as targets for nonsense suppression. Herein, we focus on the characterization of the melanocortin-4-receptor (MC4R) nonsense allele W16X identified in obese subjects. Cell culture assays revealed a loss-of-function of Mc4rX16 characterized by impaired surface expression and defect signaling. The aminoglycoside G-418 restored Mc4rX16 function in vitro demonstrating that Mc4rX16 is susceptible to nonsense suppression. For the evaluation of nonsense suppression in vivo, we generated a Mc4rX16 knock-in mouse line by gene targeting. Mc4rX16 knock-in mice developed hyperphagia, impaired glucose tolerance, severe obesity and an increased body length demonstrating that this new mouse model resembles typical characteristics of Mc4r deficiency. In a first therapeutic trial, the aminoglycosides gentamicin and amikacin induced no amelioration of obesity. Further experiments with Mc4rX16 knock-in mice will be instrumental to establish nonsense suppression for Mc4r as an obesity-associated target gene expressed in the central nervous system.

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Acknowledgements

This work was supported by the Bundesministerium für Bildung und Forschung within the framework of the National Genome Research Network NGFNplus to Martin Klingenspor (01GS0822), Wolfgang Wurst (01GS0823) and to Heike Biebermann (01GS0825). We also would like to thank the BMBF/DLR for their support to Wolfgang Wurst (01GS0858).

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Authors and Affiliations

  1. Technische Universität München, Molecular Nutritional Medicine, Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, Freising, Germany

    F Bolze, N Rink, S Mocek, A-E Schwarz, C Kless, J Rozman & M Klingenspor

  2. Institute of Experimental Pediatric Endocrinology, Charité-Universitätsmedizin Berlin, Berlin, Germany

    H Brumm & H Biebermann

  3. Institute of Developmental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg/München, Germany

    R Kühn & W Wurst

  4. Institute of Experimental Genetics, Helmholtz Zentrum München, German Research Center for Environmental Health (GmbH), Neuherberg/München, Germany

    A-E Schwarz & J Rozman

  5. Max Planck Institute of Psychiatry, München, Germany

    W Wurst

  6. Technische Universität München, Chair of Developmental Genetics, Neuherberg/München, Germany

    W Wurst

  7. DZNE, Deutsches Zentrum für Neurodegenerative Erkrankungen, LMU, Ludwig Maximilian Universität, Adolf Butenandt-Institut für Biochemie, München, Germany

    W Wurst

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  1. F Bolze
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Correspondence to F Bolze.

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Bolze, F., Rink, N., Brumm, H. et al. Characterization of the melanocortin-4-receptor nonsense mutation W16X in vitro and in vivo. Pharmacogenomics J 13, 80–93 (2013). https://doi.org/10.1038/tpj.2011.43

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