Characterisation of a variant linked to cortisol-producing adrenocortical adenoma
Jamaluddin, Aqfan Wyatt, Rachael Pasaliu, Andreea Ruggles, Oliver Calebiro, Davide Gorvin, Caroline M Ronchi, Cristina L
Jamaluddin, Aqfan
Wyatt, Rachael
Pasaliu, Andreea
Ruggles, Oliver
Calebiro, Davide
Gorvin, Caroline M
Ronchi, Cristina L
Affiliation
University of Birmingham; University of Nottingham; University Hospitals Birmingham NHS Foundation Trust; Birmingham Health Partners
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Publication date
2025-05-16
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Abstract
Objective: Adrenocortical adenomas are frequent in the general population and can be associated with autonomous cortisol excess, increasing morbidity and mortality. Altered cAMP/PKA signalling is common in sporadic cortisol-producing adenomas, typically due to somatic activating mutations in the catalytic subunit α of PKA (PRKACA) or the G-protein α subunit, Gαs (GNAS), which activate cAMP signalling. We previously identified a novel p.Lys58Gln GNAS somatic variant in a patient with a 5.3 cm adenoma and overt Cushing's syndrome. This novel mutation was not charactersised before but provided enough evidence to warrant further investigation.
Design and methods: Using HEK293 cells depleted of GNAS, we established wild-type (WT) Gαs and Gαs-Lys58Gln stable cell lines and evaluated adrenocorticotropic hormone (ACTH) receptor signalling using a cAMP GloSensor assay, measured CREB transcription factor phosphorylation (pCREB) by AlphaLISA and assessed CRE luciferase reporter activity. Cell viability and apoptosis were also assessed over 5 days.
Results: The Gαs-Lys58Gln variant showed a significantly higher basal cAMP, pCREB and CRE luciferase reporter concentration and a greater response to ACTH (0-10 nM, P < 0.001) compared to WT Gαs. The variant had no effect on ligand potency. There was also significantly enhanced cell viability and apoptosis in cells with the Gαs-Lys58Gln variant.
Conclusions: In conclusion, our study demonstrated that the Gαs-Lys58Gln variant is associated with constitutive activation of GNAS signalling, similar to Arg201 mutations previously reported in adrenocortical adenomas, potentially representing a new pathogenic mechanism in a subset of patients with adrenal Cushing syndrome. This variant may also affect cell proliferation and requires further study.
Citation
Jamaluddin A, Wyatt R, Pasaliu A, Ruggles O, Calebiro D, Gorvin CM, Ronchi CL. Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma. Endocr Oncol. 2025 May 16;5(1):e250009. doi: 10.1530/EO-25-0009.
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Article