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

Item Type:	Article
Title:	Characterization of the melanocortin-4-receptor nonsense mutation W16X in vitro and in vivo
Creators Name:	Bolze, F. and Rink, N. and Brumm, H. and Kühn, R. and Mocek, S. and Schwarz, A.E. and Kless, C. and Biebermann, H. and Wurst, W. and Rozman, J. and Klingenspor, M.
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 Mc4r(X16) characterized by impaired surface expression and defect signaling. The aminoglycoside G-418 restored Mc4r(X16) function in vitro demonstrating that Mc4r(X16) is susceptible to nonsense suppression. For the evaluation of nonsense suppression in vivo, we generated a Mc4r(X16) knock-in mouse line by gene targeting. Mc4r(X16) 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 Mc4r(X16) knock-in mice will be instrumental to establish nonsense suppression for Mc4r as an obesity-associated target gene expressed in the central nervous system.
Keywords:	Melanocortin-4-Receptor, Obesity, Nonsense Suppression, Aminoglycosides, Animals, Mice
Source:	Pharmacogenomics Journal
ISSN:	1470-269X
Publisher:	Nature Publishing Group
Volume:	13
Number:	1
Page Range:	80-93
Date:	February 2013
Official Publication:	https://doi.org/10.1038/tpj.2011.43
PubMed:	View item in PubMed

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