Abstract
Background
FOXL2 is a transcription factor expressed in ovarian granulosa cells. A somatic variant of FOXL2 (c.402 C > G, p.Cys134Trp) is the hallmark of adult-type granulosa cell tumours.
Methods
We generated KGN cell clones either heterozygous for this variant (MUT) or homozygous for the wild-type (WT) allele by CRISPR/Cas9 editing. They underwent RNA-Seq and bioinformatics analyses to uncover pathways impacted by deregulated genes. Cell morphology and migration were studied.
Results
The differentially expressed genes (DEGs) between WT/MUT and WT/WT KGN cells (DEGs-WT/MUT), pointed to several dysregulated pathways, like TGF-beta pathway, cell adhesion and migration. Consistently, WT/MUT cells were rounder than WT/WT cells and displayed a different distribution of stress fibres and paxillin staining. A comparison of the DEGs-WT/MUT with those found when FOXL2 was knocked down (KD) in WT/WT KGN cells showed that most DEGs-WT/MUT cells were not so in the KD experiment, supporting a gain-of-function (GOF) scenario. MUT-FOXL2 also displayed a stronger interaction with SMAD3.
Conclusions
Our work, aiming at better understanding the GOF scenario, shows that the dysregulated genes and pathways are consistent with this idea. Besides, we propose that GOF might result from an enhanced interaction with SMAD3 that could underlie an ectopic capacity of mutated FOXL2 to bind SMAD4.
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Data availability
The datasets generated and/or analysed during the current study (RNA-seq) can be accessed at [Gene Expression Omnibus, GSE225781 and GSE227040] repository.
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Acknowledgements
The authors are indebted to Emma Vidal, Lakshmi Balasubramaniam, Joseph d’Alessandro and Alexandros Glentis for their help and advices.
Funding
This work was supported by the University of Paris Cité and the Centre National de la Recherche Scientifique and by ARC (Association pour la Recherche contre le Cancer) and Les Entreprises contre le cancer.
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LH: conceptualisation, formal analysis, investigation, methodology, validation, visualisation and writing—original draft. AA: investigation. BL: investigation. CDC: investigation. RAV: conceptualisation, formal analysis, funding acquisition, project administration, supervision, validation, writing—original draft. ALT: conceptualisation, formal analysis, funding acquisition, investigation, project administration, supervision, validation, visualisation, writing—original draft.
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Herman, L., Amo, A., Legois, B. et al. A cellular model provides insights into the pathogenicity of the oncogenic FOXL2 somatic variant p.Cys134Trp. Br J Cancer (2024). https://doi.org/10.1038/s41416-024-02613-x
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DOI: https://doi.org/10.1038/s41416-024-02613-x