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Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside

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 tr...

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Autores principales: Höpfner, Friederike, Paisdzior, Sarah, Reininghaus, Nanina, Sohail, Iqra, Scheerer, Patrick, Annibale, Paolo, Biebermann, Heike, Kühnen, Peter
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9697516/
https://www.ncbi.nlm.nih.gov/pubmed/36362948
http://dx.doi.org/10.3390/life12111793
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author Höpfner, Friederike
Paisdzior, Sarah
Reininghaus, Nanina
Sohail, Iqra
Scheerer, Patrick
Annibale, Paolo
Biebermann, Heike
Kühnen, Peter
author_facet Höpfner, Friederike
Paisdzior, Sarah
Reininghaus, Nanina
Sohail, Iqra
Scheerer, Patrick
Annibale, Paolo
Biebermann, Heike
Kühnen, Peter
author_sort Höpfner, Friederike
collection PubMed
description 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, G(s) 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.
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spelling pubmed-96975162022-11-26 Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside Höpfner, Friederike Paisdzior, Sarah Reininghaus, Nanina Sohail, Iqra Scheerer, Patrick Annibale, Paolo Biebermann, Heike Kühnen, Peter Life (Basel) Article 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, G(s) 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. MDPI 2022-11-05 /pmc/articles/PMC9697516/ /pubmed/36362948 http://dx.doi.org/10.3390/life12111793 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Höpfner, Friederike
Paisdzior, Sarah
Reininghaus, Nanina
Sohail, Iqra
Scheerer, Patrick
Annibale, Paolo
Biebermann, Heike
Kühnen, Peter
Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside
title Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside
title_full Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside
title_fullStr Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside
title_full_unstemmed Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside
title_short Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside
title_sort evaluation of pharmacological rescue of melanocortin-4 receptor nonsense mutations by aminoglycoside
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9697516/
https://www.ncbi.nlm.nih.gov/pubmed/36362948
http://dx.doi.org/10.3390/life12111793
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