Abstract
In ovarian carcinoma, anti-Müllerian hormone (AMH) type II receptor (AMHRII) and the AMH/AMHRII signaling pathway are potential therapeutic targets. Here, AMH dose-dependent effect on signaling and proliferation was analyzed in four ovarian cancer cell lines, including sex cord stromal/granulosa cell tumors and high grade serous adenocarcinomas (COV434-AMHRII, SKOV3-AMHRII, OVCAR8 and KGN). As previously shown, incubation with exogenous AMH at concentrations above the physiological range (12.5–25 nM) decreased cell viability. Conversely, physiological concentrations of endogenous AMH improved cancer cell viability. Partial AMH depletion by siRNAs was sufficient to reduce cell viability in all four cell lines, by 20% (OVCAR8 cells) to 40% (COV434-AMHRII cells). In the presence of AMH concentrations within the physiological range (5 to 15 pM), the newly developed anti-AMH B10 antibody decreased by 25% (OVCAR8) to 50% (KGN) cell viability at concentrations ranging between 3 and 333 nM. At 70 nM, B10 reduced clonogenic survival by 57.5%, 57.1%, 64.7% and 37.5% in COV434-AMHRII, SKOV3-AMHRII, OVCAR8 and KGN cells, respectively. In the four cell lines, B10 reduced AKT phosphorylation, and increased PARP and caspase 3 cleavage. These results were confirmed in ovarian cancer cells isolated from patients’ ascites, demonstrating the translational potential of these results. Furthermore, B10 reduced COV434-MISRII tumor growth in vivo and significantly enhanced the median survival time compared with vehicle (69 vs 60 days; p = 0.0173). Our data provide evidence for a novel pro-survival autocrine role of AMH in the context of ovarian cancer, which was targeted therapeutically using an anti-AMH antibody to successfully repress tumor growth.
Highlights
We analyzed the effect of anti-Müllerian hormone (AMH) concentration on cell survival in four AMHRII-positive ovarian cancer cell lines, COV434-AMHRII, SKOV3-AMHRII, OVCAR8, and KGN, using an MTS assay to measure cell viability and proliferation
Cell viability was significantly increased by the lowest tested concentrations of active recombinant human AMH (0.8 nM LR-AMH), whereas it was reduced by incubation with LR-AMH doses above the physiological range (Fig. 1a)
To analyze the involvement of AMH non-canonical signaling p athways[15,16], we monitored AKT phosphorylation and found that it was decreased upon incubation with LR-AMH, as observed for cell viability (Fig. 1b)
Summary
Production and purification of recombinant AMH is still a c hallenge[12], Pépin et al recently described an original production strategy and an alternative delivery approach using gene therapy to be translated in clinical phase[13,14] To overcome this challenge and to determine whether lower doses of AMH could be used, we wanted to identify the lowest AMH concentration that can induce apoptosis and tumor regression. The effect on proliferation/viability was inhibited by anti-AMH siRNAs and by the new anti-AMH monoclonal antibody B10 characterized in this study This antibody reduced cell viability and increased apoptosis in four ovarian cancer cell lines and in ascites cells from patients with ovarian cancer. Our data provide a new vision of AMH in the context of ovarian cancer and open the way to an innovative therapeutic approach to suppress AMH proliferative effect
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