Ovarian granulosa cell tumors (GCTs) are indolent tumors of the ovary affecting women at all ages and potentially displaying late recurrence. Even if there is still little information regarding the mechanisms involved in GCT development and progression, FOXL2 would be a major tumor suppressor gene in granulosa cells. We analyzed the mechanisms underlying GCT initiation and progression by using mice with targeted expression of SV40 large T-antigen in granulosa cells (AT mouse), which develop GCTs. Consistent with patients, AT mice with developing GCTs displayed increased levels in circulating anti-Müllerian hormone (AMH), estradiol and androgens, as well as decreased FOXL2 protein abundance. Very few mice developed metastases (1 out of 30). In situ analyses revealed that GCT initiation resulted from both increased granulosa cell survival and proliferation in large antral follicles. Tumorigenesis was associated with the combined inactivation of p53 and Rb pathways, as shown by the impaired expression of respective downstream targets regulating cell apoptosis and proliferation, i.e., Bax, Bak, Gadd45a, Ccna2, Ccne1, E2f1, and Orc1. Importantly, the expression of FOXL2 was still present in newly developed GCTs and its downregulation only started during GCT growth. Collectively, our experiments provide evidence that disrupted p53/Rb signaling can drive tumor initiation and growth. This model challenges the current paradigm that impaired FOXL2 signaling is a major switch of granulosa cell tumorigenesis, albeit possibly contributing to tumor growth.
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We wish to thank Pr. R Veitia, Dr. AL Todeschini (Institut Jacques Monod, Paris, France), and Dr. S Bouret (Institut du Cerveau et de la Moëlle, Paris, France) for valuable discussions regarding this work. They acknowledge the technical assistance of M. Surenaud for gonadotropin assays (IMRB-INSERM U955, Hôpital Henri Mondor, France), and Dr. B. Querat and Dr. D. L’hôte (University Paris 7, BFA unit, France) for providing p53 antibody and Foxl2 primers. They also thank the technical help of Dr. M. Pannetier (INRA, Jouy en Josas, France) for the detection of FOXL2 by Western blots. They thank O. Trassard (UMS-32 INSERM, Hôpital du Kremlin Bicêtre, France) for his kind help in scanning histological slides. They acknowledge the great help of the technicians of the Buffon Animal Facility (University Paris Diderot, France). This work was supported by Ligue contre le Cancer Ile-de-France (attributed to NC), Projet Fondation ARC (PJA 20151203391, attributed to CJG) and GEFLUC Paris-Ile de France (CJG). It was also funded by “Institut National de la Santé & de la Recherche Médicale” (Inserm), “Center National de la Recherche Scientifique” (CNRS), University Paris Diderot, and by fellowships from Ecole Doctorale Bio-SPC (VC, MMD, CMF).
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Cluzet, V., Devillers, M.M., Petit, F. et al. Aberrant granulosa cell-fate related to inactivated p53/Rb signaling contributes to granulosa cell tumors and to FOXL2 downregulation in the mouse ovary. Oncogene 39, 1875–1890 (2020). https://doi.org/10.1038/s41388-019-1109-7