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. 2021 Jan 26;11(1):2231.
doi: 10.1038/s41598-021-81819-y.

Anti-Müllerian hormone (AMH) autocrine signaling promotes survival and proliferation of ovarian cancer cells

Maëva Chauvin  1   2   3   4   5 , Véronique Garambois  1   2   3   4 , Pierre-Emmanuel Colombo  1   2   3   4 , Myriam Chentouf  1   2   3   4 , Laurent Gros  1   2   3   4 , Jean-Paul Brouillet  1   2   3   4   6 , Bruno Robert  1   2   3   4 , Marta Jarlier  1   2   3   4 , Karen Dumas  7 , Pierre Martineau  1   2   3   4 , Isabelle Navarro-Teulon  1   2   3   4 , David Pépin  8   5 , Thierry Chardès  1   2   3   4 , André Pèlegrin  9   10   11   12
Affiliations

Anti-Müllerian hormone (AMH) autocrine signaling promotes survival and proliferation of ovarian cancer cells

Maëva Chauvin et al. Sci Rep. .

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.

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Conflict of interest statement

Maëva Chauvin, Myriam Chentouf, Pierre Martineau, Bruno Robert, and André Pèlegrin are inventors of a patent “ANTI-MÜLLERIAN INHIBITING SUBSTANCE ANTIBODIES AND USES THEREOF” filed on 2019, September, 27. Maëva Chauvin, Thierry Chardès, Isabelle Navarro-Teulon and André Pèlegrin are inventors of a patent “USE OF MÜLLERIAN INHIBITING SUBSTANCE INHIBITORS FOR TREATING CANCER” filed on 2019, September, 27. David Pépin is inventor of the patent “MODIFIED MULLERIAN INHIBITING SUBSTANCE (MIS) PROTEINS AND USES THEREOF FOR THE TREATMENT OF DISEASES”, WO2014164981A1. The other authors declare no competing interests.

Figures

Figure 1
Figure 1
Low-dose recombinant AMH (LR-AMH) promotes cell viability in COV434-AMHRII, SKOV3-AMHRII, OVCAR8 and KGN cells. (a) Cell viability (MTS assay) and (b) AKT phosphorylation (western blotting) after incubation with 0.8 to 25 nM LR-AMH for 3 days. (c) AMH expression at 48 h after transfection of siRNAs against AMH, and (d) Effect of siRNA-mediated AMH silencing on cell viability at day 3 post-transfection. Figure shows a representative experiment out of 2 to 4 experiments performed depending on the cell line (mean ± SEM of 3 replicate wells) *p < 0.05; ***p < 0.001. For western blot figure preparation, all proteins were analyzed on the same gel and membrane. After blotting, the lines corresponding to the different proteins were cut and may have been exposed for different lengths of time because the objective was not to compare the different proteins with each other, but to see the effect of AMH (b) or of siRNA (c) on the expression of each protein.
Figure 2
Figure 2
The anti-AMH antibody B10 reduces cell viability/proliferation and induces growth inhibition in COV434-AMHRII, SKOV3-AMHRII, OVCAR8, and KGN cells. (a) Cell viability was analyzed (MTS assay) after incubation with 3 to 333 nM B10 for 3 days. (b) Clonogenic survival in COV434-AMHRII cells (direct clone counting) and in SKOV3-AMHRII, OVCAR8, and KGN cells (measurement of cell confluence using the Celigo Imaging System) after incubation or not with 70 nM B10 for 11 days. (c) AKT phosphorylation and apoptosis induction (cleaved caspase 3 and PARP) after incubation or not with 333 nM B10 for 24 h. All proteins were analyzed on the same gel and membrane. After blotting, the lines corresponding to the different proteins were cut and may have been exposed for different lengths of time because the objective was not to compare the different proteins with each other, but to see the effect of B10 on the expression of each protein. Figure shows a representative experiment out of 2 to 4 experiments performed depending on the cell line (mean ± SEM of 3 replicate wells) *p < 0.05.
Figure 3
Figure 3
The anti-AMH antibody B10 reduces cell viability/proliferation and induces growth inhibition in tumor cells from ovarian carcinoma ascites samples. (a) Cell viability analyzed with the MTS assay after incubation with 0.3 to 333 nM B10 for 3 days. (b) Cell growth inhibition (cell confluence measured with the Celigo Imaging System) after incubation or not (NT) with 70 nM B10 for 48 h. (c) Apoptosis induction (caspase 3/7 activity) after incubation or not with increasing concentrations of B10. Figure shows mean ± SEM of 3 replicate wells. *p < 0.05; **p < 0.01; ***p < 0.001.
Figure 4
Figure 4
The anti-AMH antibody B10 reduces COV434-MISRII tumor growth in vivo. Nude mice bearing COV434-MISRII cell tumors were treated with B10 (anti-AMH antibody), 12G4 (anti-AMHRII antibody) (10 mg/kg/injection for both), or vehicle (NaCl; control) twice a week for 4 weeks. (a) Tumor growth curves (mean + 95% confidence intervals), and (b) Kaplan–Meier survival curves (percentage of mice with a tumor volume < 1500 mm3 as a function of time after graft).
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References

    1. di Clemente N, et al. Processing of anti-mullerian hormone regulates receptor activation by a mechanism distinct from TGF-beta. Mol. Endocrinol. 2010;24:2193–2206. doi: 10.1210/me.2010-0273. - DOI - PMC - PubMed
    1. Josso N, et al. Anti-Müllerian hormone in early human development. Early Hum. Dev. 1993;33:91–99. doi: 10.1016/0378-3782(93)90204-8. - DOI - PubMed
    1. Horbelt D, Denkis A, Knaus P. A portrait of transforming growth factor β superfamily signalling: Background matters. Int. J. Biochem. Cell Biol. 2012;44:469–474. doi: 10.1016/j.biocel.2011.12.013. - DOI - PubMed
    1. Xavier F, Allard S. Anti-Müllerian hormone, beta-catenin and Müllerian duct regression. Mol. Cell. Endocrinol. 2003;211:115–121. doi: 10.1016/j.mce.2003.09.022. - DOI - PubMed
    1. McLennan IS, Pankhurst MW. Anti-Müllerian hormone is a gonadal cytokine with two circulating forms and cryptic actions. J. Endocrinol. 2015;226:R45–57. doi: 10.1530/JOE-15-0206. - DOI - PubMed

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