Bladder cancer (BCa) is the sixth most common cancer in the United States. Pathologically, BCa is categorized into muscle invasive (MIBC) and non-muscle invasive (NMIBC) disease. NMIBC accounts for approximately 75% of BCa cases. While the majority of NMI tumors carry a good prognosis; recurrence is common with progression to MI and metastatic disease. BCa requires life-long surveillance, thus BCa is the one of the most expensive cancer to treat. Treatment for BCa remained stagnant in the last 30 years with a few new-approved drugs for advanced disease. Thus, there is unmet need for better understanding of the pathobiology of the disease to develop better therapeutics. Metabolic programming of cancer is increasingly recognized as one of the hallmarks of cancer as cancer cells exploit multiple pathways to generate ATP and metabolic intermediates for the biomass of the rapidly proliferating, invasive and resistant cells. To date, profiling of metabolic programming of BCa is still in infancy. To bridge this knowledge gap, we performed comprehensive metabolomics and transcriptomic profiling of urothelial lesions isolated from the well-established murine chemical carcinogenesis model integrated with patients' data. We validated the therapeutic potential of targeting the upregulated metabolic programs in preclinical models. We found that early urothelial carcinogenesis is associated with progressively increased levels of glucose and glycolysis intermediates concomitant with shift to pentose phosphate pathway with increased levels with AMP and nucleotide precursors required for ATP production and redox homeostasis. Early urothelial lesions also exhibited significant progressive increase in carnitine-conjugated fatty acids suggesting increased fatty acid synthesis, oxidation or both. Transcriptomic profiling of patients' tumors across multiple datasets revealed significant upregulation of glycolytic enzymes in tumors compared to normal tissues, with negative correlation of glucose transporters and glycolytic enzymes with patients' survival. The enzymes regulating both fatty acid synthesis and oxidation are not only upregulated in patients' tumors but are also associated with poor survival. To further determine metabolic dependence/vulnerability of BCa, we treated BCa cell lines with inhibitors of glycolysis (2-DG), ATP production (AMP analog AICAR), TCA cycle (CPI-613), fatty acid synthesis (orlistat), fatty acid oxidation (etomoxir) and mitochondrial electron transport chain (phenformin). We found that each of these inhibitors exhibited significant dose-dependent inhibition on BCa cell survival and invasiveness in vitro. Together, our data provide evidence of metabolic plasticity BCa and dependence on multiple energy generating pathways that be exploited as potential therapeutic targets.

Citation Format: Hesham Afify, Daniela Gonzalez, Alia Ghoneum, Neveen A. Said. Metabolic programming of bladder cancer: Vulnerabilities and opportunities [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4772.