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Genetics and Genomics

A cellular model provides insights into the pathogenicity of the oncogenic FOXL2 somatic variant p.Cys134Trp

British Journal of Cancer (2024)Cite this article

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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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Fig. 1: Profound transcriptomic changes due to the presence of the C134W pathogenic variant point to a perturbation of TGFb signalling.
Fig. 2: Comparison of DEGs in WT/MUT KGN cells or mAGCTs and DEGs in the ovary of the Trim28 KO mouse.
Fig. 3: Comparison of DEGs-WT/MUT (KGN cells) versus hAGCTs or mAGCTs.
Fig. 4: Interplay of DEGs-WT/MUT (KGN cells) with TADs and ChIPseq peaks.
Fig. 5: Morphological differences of WT/MUT vs WT/WT KGN cells.
Fig. 6: Stress fibres and focal adhesions in WT/MUT and WT/WT KGN cells.
Fig. 7: TGF-β/Smad signalling in WT/MUT cells.

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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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Author notes

  1. These authors jointly supervised this work: Reiner A. Veitia, Anne-Laure Todeschini.

Authors and Affiliations

  1. Université Paris Cité, CNRS, Institut Jacques Monod, Paris, France

    Laetitia Herman, Angélique Amo, Berangère Legois, Caterina Di Carlo, Reiner A. Veitia & Anne-Laure Todeschini

  2. Université Paris Saclay, Paris, France

    Reiner A. Veitia

  3. Institut de Biologie François Jacob, CEA, Fontenay aux Roses, Paris, France

    Reiner A. Veitia

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Contributions

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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Correspondence to Anne-Laure Todeschini.

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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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