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

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Item Type:	Article
Title:	Characterization of the melanocortin-4-receptor nonsense mutation W16X in vitro and in vivo
Creators Name:	Bolze, F., Rink, N., Brumm, H., Kühn, R., Mocek, S., Schwarz, A.E., Kless, C., Biebermann, H., Wurst, W., 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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