Abstract 2020: Investigation of CYP450 enzymes as therapeutic targets in rhabdomyosarcoma

Abstract

Abstract Introduction: Rhabdomyosarcoma (RMS) is a rare soft tissue sarcoma (STS) that predominantly affects children and teenagers. It is the most frequent STS in children and accounts for around 5-8% of total childhood cancers. The improvement of chemotherapeutic regimens has raised RMS survival chances over the last decades. However, the mortality rate remains exceptionally high in high-risk RMS patients. Duocarmycins have arisen as an opportunity for targeted therapy in cancer. These ultrapotent anticancer agents have been reengineered to generate bioprecursors that are selectively activated bycertain cytochrome P450 (CYP) isoforms (e.g. CYP1A1, CYP1B1, CYP2W1). The overexpression of these isoforms in tumors provides the possibility to locoregionally bioactivate duocarmycin prodrugs only in the tumor site. Previous studies have shown that duocarmycins can be chemically modified and deactivated by removal of a hydroxyl group in the pharmacophore, which can be reinstalled by CYPsto generate a potent DNA-targeted cytotoxic metabolite. In the past, a consistent CYP2W1overexpression in patient-derived RMS biopsies as compared to matched, non-transformed tissue was revealed. Results: Ongoing studies are focused on measuring CYP1A1, CYP1B1 and CYP2W1 expression levels in RMS cell lines and in three-dimensional spheroids as well as evaluating the antiproliferative activity of duocarmycin bioprecursors in these models. With the exception of RH30 cells (IC50=0.2µM forICT2700) the IC50 for CYP1A1- and CYP2W1-bioactivated duocarmycins (ICT2700 and ICT2706,respectively) lie in the low micromolar range in RMS monolayer cultures, suggesting that CYP1A1 andCYP2W1 might not be a suitable target in these 2D cancer models. Consistently, only RH30 cells displayed a high CYP1A1 expression in monolayer cultures among the RMS cell line panel, and noCYP2W1 expression could be identified. However, preliminary data suggests that CYP1A1 and CYP2W1enzymes might be restored when spheroids are generated from RMS cell lines. A functional 7-ethoxyresorufin-O-deethylase (EROD) assay has been validated to analyze CYP1A1- and CYP1B1-specific enzymatic activity in RMS cell lines. Vorinostat (SAHA) is an epigenetic drug currently used in the clinic for cancer treatment, and it has been associated with modulatory effects on CYP1expression. Our data suggests cell line-dependent synergistic or antagonistic effects of SAHA when combined with ICT2700, and these correlate with SAHA-induced CYP1A1 upregulation and downregulation respectively at both gene and protein levels. Conclusion: Evaluation of CYP target expression in RMS is being established to assess if duocarmycin bioprecursorscan be progressed as a novel type of therapy in RMS. Interestingly, CYP450 expression in RMS cell lines seems to vary depending on the complexity of the in vitro model used. Citation Format: Enric Arasanz Picher, Klaus Pors. Investigation of CYP450 enzymes as therapeutic targets in rhabdomyosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2020.