PD51-06 TARGETING GLUTAMINE ADDICTION TO INDUCE FERROPTOSIS WITH POTENT DRUG COMBINATION THERAPY FOR RENAL CELL CARCINOMA

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology II (PD51)1 May 2024PD51-06 TARGETING GLUTAMINE ADDICTION TO INDUCE FERROPTOSIS WITH POTENT DRUG COMBINATION THERAPY FOR RENAL CELL CARCINOMA Akihito Takeuchi Akihito TakeuchiAkihito Takeuchi View All Author Informationhttps://doi.org/10.1097/01.JU.0001009360.84490.42.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: In recent years, glutaminolysis has been implicated in tumor progression in a variety of malignancies. Alanine-serine-cysteine transporter 2 (ASCT2) has been reported to be associated with progression and survival in RCC. On the other hand, our previous study examined the effect of V9302 treatment, a specific inhibitor of ASCT2, which unexpectedly resulted in increased intracellular glutamine levels. In this study, we investigated whether the combination of CB839, an inhibitor of glutaminase (GLS), which is also associated with glutaminolysis, would have a more effective antitumor effect. METHODS: The human RCC cell lines, 786-O, and 769-P were used. V9302 was used as the specific ASCT2 inhibitor and CB839 as the specific GLS inhibitor. Cell viability was assessed using WST-8 assay. The protein expression was detected by Western blotting. Intracellular metabolite concentrations were analyzed using liquid chromatography mass spectrometry (LC/MS). Reactive Oxygen Species (ROS) and lipid peroxides involved in ferroptosis were measured by flow cytometry. As the xenograft model in vivo, athymic nude mice were injected subcutaneously with 786-O cells and given various doses of V9302 and CB839. RESULTS: In both cell lines, a decrease in cell viability was observed with the combination of CB839 and V9302 compared to V9302 alone. LC/MS assays showed that intracellular glutamine levels in RCC cell lines were significantly reduced with the combination of V9302 and CB839 compared to monotherapy. In addition, glutathione levels, which are involved in the regulation of ferroptosis, were markedly decreased by combination treatment, while ROS and lipid peroxide levels were dramatically increased. In a tumor xenograft study, the combination of V9302 (30 mg/kg, intraperitoneal injection) and CB839 (200 mg/kg, oral administration) significantly inhibited tumor growth compared to the control group within 4 weeks of administration. CONCLUSIONS: The results of this study indicate that the combination of V9302 and CB839 induces potent glutamine metabolic inhibition, resulting in a more potent antitumor effect. Our results also suggest that the suppression of glutathione production through the regulation of glutamine metabolism may lead to anti-tumor effects related to ferroptosis. This combination therapy might have the potential to become a new treatment for renal cell carcinoma. Source of Funding: None © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1066 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Akihito Takeuchi More articles by this author Expand All Advertisement PDF downloadLoading ...