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

Friederike Höpfner, Sarah Paisdzior, Nanina Reininghaus, Iqra Sohail, Patrick Scheerer, Paolo Annibale, Heike Biebermann, Peter Kühnen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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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 Gq/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, Gs activation was almost impossible in W258X and Q307X with wild-type-like cell surface expression. Results for Gq/11 signaling were comparable. Based on our data, this approach improbably represents a therapeutic option.

Original languageEnglish
Article number1793
Number of pages17
JournalLife
Volume12
Issue number11
DOIs
Publication statusPublished - 5 Nov 2022

Keywords

  • Melanocortin 4 receptor
  • MC4R
  • Stop mutation
  • PTC
  • Translational readthrough
  • G418

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