Increased HRD score in cisplatin resistant penile cancer cells

Penile cancer is a rare disease in demand for new therapeutic options. Frequently used combination chemotherapy with 5 fluorouracil (5-FU) and cisplatin (CDDP) in patients with metastatic penile cancer mostly results in the development of acquired drug resistance. Availability of cell culture models with acquired resistance against standard therapy could help to understand molecular mechanisms underlying chemotherapy resistance and to identify candidate treatments for an efficient second line therapy.

We generated a cell line from a humanpapilloma virus (HPV) negative penile squamous cell carcinoma (UKF-PEC-1). This cell line was subject to chronic exposure to chemotherapy with CDDP and / or 5-FU to induce acquired resistance in the newly established chemo-resistant sublines (PEC-1^rCDDP²⁵⁰⁰, adapted to 2500 ng/ml CDDP; UKF-PEC-1^r5-FU⁵⁰⁰, adapted to 500 ng/ml 5- FU; UKF-PEC1^rCDDP²⁵⁰⁰/^r5-FU⁵⁰⁰, adapted to 2500 ng/ml CDDP and 500 ng/ml 5 -FU). Afterwards cell line pellets were formalin-fixed, paraffin embedded and subject to sequencing as well as testing for homologous recombination deficiency (HRD). Additionally, exemplary immunohistochemical stainings for p53 and gammaH2AX were applied for verification purposes. Finally, UKF-PEC-1^rCDDP²⁵⁰⁰, UKF-PEC-1^r5-FU⁵⁰⁰, UKF-PEC1^rCDDP²⁵⁰⁰/^r5-FU⁵⁰⁰, and UKF-PEC-3 (an alternative penis cancer cell line) were tested for sensitivity to paclitaxel, docetaxel, olaparib, and rucaparib.

The chemo-resistant sublines differed in their mutational landscapes. UKF-PEC-1^rCDDP²⁵⁰⁰ was characterized by an increased HRD score, which is supposed to be associated with increased PARP inhibitor and immune checkpoint inhibitor sensitivity in cancer. However, UKF-PEC-1^rCDDP²⁵⁰⁰ did not display sensitivity to PARP inhibitors.