Dual mTORC1/2 inhibition sensitizes testicular cancer models to cisplatin treatment.

Testicular cancer (TC) is the most common cancer type among young men. Despite highly effective cisplatin-based chemotherapy, around 20% of patients with metastatic disease will still die from the disease. The aim of this study was to explore the use of kinase inhibitors to sensitize testicular cancer cells to cisplatin treatment. Activation of kinases, including receptor tyrosine kinases, and downstream substrates was studied in five cisplatin-sensitive or resistant TC cell lines using phospho-kinase arrays and western blotting. The phospho-kinase array showed AKT and S6 to be among the top phosphorylated proteins in TC cells, which are part of the PI3K/AKT/mTORC pathway. Inhibitors of most active kinases in the PI3K/AKT/mTORC pathway were tested using apoptosis assays and survival assays. Two mTORC1/2 inhibitors, AZD8055 and MLN0128, strongly enhanced cisplatin-induced apoptosis in all tested TC cell lines. Inhibition of mTORC1/2 blocked phosphorylation of the mTORC downstream proteins S6 and 4E-BP1. Combined treatment with AZD8055 and cisplatin led to reduced clonogenic survival of TC cells. Two TC patient-derived xenografts (PDX), either from a chemo-sensitive or -resistant patient, were treated with cisplatin in the absence or presence of kinase inhibitor. Combined AZD8055 and cisplatin treatment resulted in effective mTORC1/2 inhibition, increased caspase-3 activity, and enhanced tumor growth inhibition. In conclusion, we identified mTORC1/2 inhibition as an effective strategy to sensitize TC cell lines and PDX models to cisplatin treatment. Our results warrant further investigation of this combination therapy in the treatment of TC patients with high risk relapsed or refractory disease.