Drug repurposing of bladder cancer driven by patients’ proteomic signatures

Abstract

Molecular signatures may present sources of druggable targets not adequately explored yet. The objective of this study was the identification of known de-risked potential therapeutic compounds (drug repurposing) for the treatment of high-risk NMIBC and MIBC, based on high resolution proteomics profiles. The next generation Connectivity Map (CMap) was employed for drug repurposing using as input proteomic signatures from patients with MIBC, and NMIBC of low (NPS3) and high aggressive molecular profile (NPS1) from our recently published datasets (Stroggilos et al. Int J Cancer.2020;146(1):281). The retrieved candidate drugs were ranked according to the disease–drug connectivity score. The impact of selected drugs was investigated in vitro in a panel of multi-origin BC cell lines including benign (HBLAK), non–muscle invasive (BFTC-905, SW1710) and invasive (T24, T24M, VmCub1, 253J, HT-1376) cells. The impact on cell proliferation (MTS), colony formation (matrigel), and apoptosis (Annexin V) was assessed. Among the top ranked compounds reversing the molecular signature of NMIBC from high to low risk subtype were mTOR inhibitors, tubulin inhibitors, caspase activators and RAF inhibitors. WYE-354, an ATP-competitive inhibitor of mTOR, had the highest connectivity score. WYE-354 was also ranked among the top candidate drugs when using as input proteins with consistent changes at the mRNA level in NMIBC progressors versus non-progressors from earlier published studies or proteomics changes in MIBC versus NMIBC. In vitro administration of WYE-354 resulted in a significant reduction in the proliferation rate of all the tested BC cell lines in a concentration-dependent manner and also impaired colony growth by significantly decreasing the size of the colonies. However, no significant effect on BC cell apoptosis could be observed. In this study, we describe a promising pipeline for identifying repurposed drugs of potential therapeutic impact for aggressive NMIBC, based on patients’ proteomic signatures. Deciphering the molecular mechanism of the impact of WYE-354 based on the molecular characteristics of the tested cell lines is ongoing. Evaluation of further compounds or combinations predicted by the CMap analysis is also planned.