ADAM17 Inhibition Increases the Impact of Cisplatin Treatment in Ovarian Cancer Spheroids.

Nina Hedemann,Andreas Herz,Susanne Sebens,Jan Dittrich,Christoph Rogmans,Sandra Krüger,Julia Feuerborn,Nils Tribian,Jörg Weimer,Jan Hendrik Schiepanski,Nicolai Maass,Dirk O Bauerschlag,Inken Flörkemeier,Astrid Dempfle

doi:10.3390/cancers13092039

Abstract

Simple SummaryOvarian cancer (OvCa) treatment is still a challenge, mainly due to acquired resistance mechanisms during the course of chemotherapy. Here, we show the enhanced cytotoxicity of the combined treatment with the ADAM17 inhibitor GW280264X and cisplatin in comparison with cisplatin monotherapy. This effect was visible in five of five ovarian cancer cell lines grown as a monolayer and two of three tested cell lines in three-dimensional tumor spheroids. Tumor spheroids derived from primary tumor and ascites cells were sensitized to cisplatin treatment by GW280264X. In summary, the combination of ADAM17 inhibition with conventional chemotherapy seems to be a promising strategy to overcome chemotherapy resistance in OvCa.Chemotherapy resistance is a major challenge in ovarian cancer (OvCa). Thus, novel treatment combinations are highly warranted. However, many promising drug candidates tested in two-dimensional (2D) cell culture have not proved successful in the clinic. For this reason, we analyzed our drug combination not only in monolayers but also in three-dimensional (3D) tumor spheroids. One potential therapeutic target for OvCa is A disintegrin and metalloprotease 17 (ADAM17). ADAM17 can be activated by chemotherapeutics, which leads to enhanced tumor growth due to concomitant substrate cleavage. Therefore, blocking ADAM17 during chemotherapy may overcome resistance. Here, we tested the effect of the ADAM17 inhibitor GW280264X in combination with cisplatin on ovarian cancer cells in 2D and 3D. In 2D, the effect on five cell lines was analyzed with two readouts. Three of these cell lines formed dense aggregates or spheroids (HEY, SKOV-3, and OVCAR-8) in 3D and the treatment effect was analyzed with a multicontent readout (cytotoxicity, viability, and caspase3/7 activation). We tested the combined therapy on tumor spheroids derived from primary patient cells. In 2D, we found a significant reduction in the half minimal (50%) inhibitory concentration (IC50) value of the combined treatment (GW280264X plus cisplatin) in comparison with cisplatin monotherapy in all five cell lines with both 2D readout assays (viability and caspase activation). In contrast, the combined treatment only showed an IC50 reduction in HEY and OVCAR-8 3D tumor spheroid models using caspase3/7 activity or CelltoxTM Green as the readout. Finally, we found an improved effect of GW280264X with cisplatin in tumor spheroids derived from patient samples. In summary, we demonstrate that ADAM17 inhibition is a promising treatment strategy in ovarian cancer.

Highlights

Ovarian cancer (OvCa) is one of the most common gynecological malignancies [1].In the U.S, the majority (76%) of women are diagnosed in an advanced stage FédérationInternationale de Gynécologie et d’Obstétrique III and IV (FIGO III and IV) when disease has already spread within the abdominal cavity
We systematically evaluated the half minimal (50%) inhibitory concentration (IC50 ) values in five ovarian cancer (OvCa) cell lines applying a fluorescence-based viability assay 48 h after treatment
The evaluation of IC50 values revealed greatly reduced cell viability using combinatorial treatment with GW280264X (ADAM10 and A disintegrin and metalloprotease 17 (ADAM17) selective) and cisplatin compared with cisplatin-only treatment

Summary

Introduction

Ovarian cancer (OvCa) is one of the most common gynecological malignancies [1].In the U.S, the majority (76%) of women are diagnosed in an advanced stage FédérationInternationale de Gynécologie et d’Obstétrique III and IV (FIGO III and IV) when disease has already spread within the abdominal cavity. Ovarian cancer (OvCa) is one of the most common gynecological malignancies [1]. In the U.S, the majority (76%) of women are diagnosed in an advanced stage Fédération. Internationale de Gynécologie et d’Obstétrique III and IV (FIGO III and IV) when disease has already spread within the abdominal cavity. Most patients develop chemotherapy resistance during the course of therapy, which is a major drawback of current chemotherapeutics [2]. The average relative 5-year survival rate of 43% in the U.S indicates a need for more effective strategies to improve patient outcomes [3]. Over the last few decades, only a few breakthroughs have occurred regarding the treatment of OvCa. One of them was the introduction of poly-ADP-ribose-polymerase (PARP)

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