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

Abstract

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.

The Influence of Antitumor Unsymmetrical Bisacridines on 3D Cancer Spheroids Growth and Viability

“…For example, HT29 spheroids were used to determine drug responses and highlight the effects of the drugs oxaliplatin, 5-fluorouracil, and folinic acid following the encapsulation of their respective liposomes [ 35 ]. In other studies, tumor spheroids derived from ovarian cancer cell lines or primary patients’ cells were sensitized to cisplatin treatment by using an inhibitor of A disintegrin and metalloprotease 17 (ADAM17), named GW280264X, to overcome resistance [ 36 ]. In summary, the application of 3D models has the potential to improve drug discovery research and bridge the gap between results obtained in preclinical phases and promising outcomes found in clinical trials.…”

High Antitumor Activity of the Dual Topoisomerase Inhibitor P8-D6 in Breast Cancer

“…The number of cells for seeding was determined experimentally. The aim was to create a spheroid with a size of 400–600 μm after 96 h. To form spheroids, cells were maintained for 96 h, then half of the medium was removed and treated as triplicates with control and treatment groups for 48 h. Simultaneously, CellTox™ Green assay (Promega #G8731) was added and detected (485Ex/520Em) 24 h and 48 h after treatment using NYONE ®® Scientific (SYNENTEC) with 4× magnification [ 18 ]. The following excitation sources and emission filters were used: Brightfield—BFEx/GreenEm (530/43 nm); CellToxTM Green (BlueEx (475/28 nm)/GreenEm (530/43 nm)).…”

NGF/TRKA Promotes ADAM17-Dependent Cleavage of P75 in Ovarian Cells: Elucidating a Pro-Tumoral Mechanism

“…These discrepancies could be explained at least in part by the use of antibodies that recognize different aminoacidic sequences of P75 (and then the intact receptor or its fragments), and also by the interaction of P75-ICD with other receptors producing its trans-activation (for instance, TRK receptors), which could yield P75-mediated tumoral effects. Because ADAM17 displays an important role in inflammatory diseases, it has been proposed as a therapeutic target in several kinds of cancer [51], including EOC [32,52,53]. This knowledge has motivated the development of pharmacologic inhibitors and antibodies targeting ADAM17 as anti-tumoral therapy, which have been tested in some preclinical models of EOC [51][52][53].…”

“…Because ADAM17 displays an important role in inflammatory diseases, it has been proposed as a therapeutic target in several kinds of cancer [51], including EOC [32,52,53]. This knowledge has motivated the development of pharmacologic inhibitors and antibodies targeting ADAM17 as anti-tumoral therapy, which have been tested in some preclinical models of EOC [51][52][53].…”