David Holthaus ^1,2 Christoph Rogmans ¹ Ina Gursinski ¹ Alvaro Quevedo-Olmos ² Marzieh Ehsani ² Mandy Mangler ^3,4 Inken Flörkemeier ¹ Jörg P. Weimer ¹ Thomas F. Meyer ² Nicolai Maass ¹ Dirk O. Bauerschlag ^1,5 Nina Hedemann ^1*

• ¹ Department of Gynaecology and Obstetrics, University Hospital Schleswig-Holstein, Kiel, Schleswig-Holstein, Germany
• ² Laboratory of Infection Oncology, Institute of Clinical Molecular Biology, Christian-Albrechts-Universität zu Kiel and University Hospital Schleswig-Holstein, Kiel, Schleswig-Holstein, Germany
• ³ Vivantes Auguste Viktoria Clinic, Berlin, Baden-Württemberg, Germany
• ⁴ Department of Gynecology, Charité University Medicine Berlin, Berlin, Baden-Württemberg, Germany
• ⁵ Department of Gynecology and Reproduction Medicine, University Hospital Jena, Jena, Thuringia, Germany

Cervical cancer represents one of the main causes of female, cancer-related mortality worldwide. The majority of cancers are caused by human papillomaviruses such as HPV16 and HPV18. As chemotherapeutic resistance to first-line platinum treatment is still a predominant clinical challenge in advanced cervical cancer, novel treatment options including combinatorial therapies are urgently required to overcome chemotherapeutic resistance. Inhibition of A Disintegrin And Metalloproteinase (ADAM)-family members, heavily involved in tumour progression of a vast range of solid tumours, strongly improved response to chemotherapeutic treatment in other tumour entities including ovarian cancer.We established two-and three-dimensional models derived from three traditional cervical cancer cell lines and ectocervical cancer-derived organoids. Following characterisation, these models were used to investigate their response to cisplatin treatment in the absence and presence of ADAM inhibitors using viability assays and automated live cell imaging.The pivotal role of the metalloprotease ADAM17 driving chemotherapy resistance was detectable in all ectocervical cultures irrespective of the model system used, whereas ADAM10 inhibition was predominantly effective only in loosely aggregated spheroids. We showed prominent differences regarding treatment responses between 2D monolayers compared to 3D spheroid and 3D organoid model systems. Particularly, the organoid system, regarded as the closest representation of primary tumours, exhibited reliably the combinatorial effect of ADAM17 inhibition and cisplatin in all three individual donors. As two-and three-dimensional models of the same cell lines differ in their responses to chemotherapy it is essential to validate treatment strategies in more advanced model systems representing the patient situation more realistically. Ectocervical organoids showed reliable results regarding treatment responses closely mimicking the primary tumours and could therefore serve as an important tool for personalized medicine in cervical cancer. These findings strengthen the role of ADAM17 as a potential novel target for combinatorial treatments to overcome chemoresistance in cervical cancer.