Establishment and Molecular Characterization of an In Vitro Model for PARPi-Resistant Ovarian Cancer

Author:

Klotz Daniel Martin123,Schwarz Franziska Maria123ORCID,Dubrovska Anna2345ORCID,Schuster Kati123,Theis Mirko26ORCID,Krüger Alexander27,Kutz Oliver123,Link Theresa123,Wimberger Pauline123,Drukewitz Stephan278,Buchholz Frank236ORCID,Thomale Jürgen9,Kuhlmann Jan Dominik123ORCID

Affiliation:

1. Department of Gynecology and Obstetrics, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany

2. National Center for Tumor Diseases (NCT), 01307 Dresden, Germany; German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany; Faculty of Medicine, University Hospital Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany; Helmholtz-Zentrum Dresden-Rossendorf (HZDR), 01307 Dresden, Germany

3. German Cancer Consortium (DKTK), Partner Site Dresden and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany

4. Helmholtz-Zentrum Dresden-Rossendorf, Institute of Radiooncology-OncoRay, 01328 Dresden, Germany

5. OncoRay-National Center for Radiation Research in Oncology, Faculty of Medicine and University Hospital Carl Gustav Carus, Technische Universität Dresden and Helmholtz-Zentrum Dresden-Rossendorf, 01307 Dresden, Germany

6. UCC Section Medical Systems Biology, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, 01307 Dresden, Germany

7. Core Unit for Molecular Tumor Diagnostics (CMTD), National Center for Tumor Diseases (NCT), Partner Site Dresden, German Cancer Consortium (DKTK), Dresden, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany

8. Institute of Human Genetics, University of Leipzig Medical Center, 04103 Leipzig, Germany

9. Institute of Cell Biology (Cancer Research), University of Duisburg-Essen Medical School, 45147 Essen, Germany

Abstract

Overcoming PARPi resistance is a high clinical priority. We established and characterized comparative in vitro models of acquired PARPi resistance, derived from either a BRCA1-proficient or BRCA1-deficient isogenic background by long-term exposure to olaparib. While parental cell lines already exhibited a certain level of intrinsic activity of multidrug resistance (MDR) proteins, resulting PARPi-resistant cells from both models further converted toward MDR. In both models, the PARPi-resistant phenotype was shaped by (i) cross-resistance to other PARPis (ii) impaired susceptibility toward the formation of DNA-platinum adducts upon exposure to cisplatin, which could be reverted by the drug efflux inhibitors verapamil or diphenhydramine, and (iii) reduced PARP-trapping activity. However, the signature and activity of ABC-transporter expression and the cross-resistance spectra to other chemotherapeutic drugs considerably diverged between the BRCA1-proficient vs. BRCA1-deficient models. Using dual-fluorescence co-culture experiments, we observed that PARPi-resistant cells had a competitive disadvantage over PARPi-sensitive cells in a drug-free medium. However, they rapidly gained clonal dominance under olaparib selection pressure, which could be mitigated by the MRP1 inhibitor MK-751. Conclusively, we present a well-characterized in vitro model, which could be instrumental in dissecting mechanisms of PARPi resistance from HR-proficient vs. HR-deficient background and in studying clonal dynamics of PARPi-resistant cells in response to experimental drugs, such as novel olaparib-sensitizers.

Publisher

MDPI AG

Subject

Cancer Research,Oncology

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