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Evaluation of pharmacological rescue of melanocortin-4 receptor nonsense mutations by aminoglycoside

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Item Type:Article
Title:Evaluation of pharmacological rescue of melanocortin-4 receptor nonsense mutations by aminoglycoside
Creators Name:Höpfner, F. and Paisdzior, S. and Reininghaus, N. and Sohail, I. and Scheerer, P. and Annibale, P. and Biebermann, H. and Kühnen, P.
Abstract:The melanocortin-4 receptor (MC4R) is critical for central satiety regulation, therefore presenting a potent target for pharmacological obesity treatment. Melanocortin-4 receptor mutations prevalently cause monogenetic obesity. A possibility of overcoming stop mutations is aminoglycoside-mediated translational readthrough. Promising results were achieved in COS-7 cells, but data for human cell systems are still missing, so uncertainty surrounds this potential treatment. In transfected HEK-293 cells, we tested whether translational readthrough by aminoglycoside Geneticin combined with high-affinity ligand setmelanotide, which is effective in proopiomelanocortin or leptin receptor deficiency patients, is a treatment option for affected patients. Five MC4R nonsense mutants (W16X, Y35X_D37V, E61X, W258X, Q307X) were investigated. Confocal microscopy and cell surface expression assays revealed the importance of the mutations' position within the MC4R. N-terminal mutants were marginally expressed independent of Geneticin treatment, whereas mutants with nonsense mutations in transmembrane helix 6 or helix 8 showed wild-type-like expression. For functional analysis, Gs and G(q/11) signaling were measured. N-terminal mutants (W16X, Y35X_D37V) showed no cAMP formation after challenge with alpha-MSH or setmelanotide, irrespective of Geneticin treatment. Similarly, G(s) activation was almost impossible in W258X and Q307X with wild-type-like cell surface expression. Results for G(q/11) signaling were comparable. Based on our data, this approach improbably represents a therapeutic option.
Keywords:Melanocortin 4 Receptor, MC4R, Stop Mutation, PTC, Translational Readthrough, G418
Page Range:1793
Date:5 November 2022
Official Publication:https://doi.org/10.3390/life12111793
PubMed:View item in PubMed

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