To develop the drug repurposing pipeline for BC, we initially focused on the proteomic signature characterizing aggressive NMIBC based on our previously published high-resolution tissue proteomics analyses. This signature was utilized as input data in the CMap tool (
https://clue.io/(si apre in una nuova finestra)) for data-driven drug repurposing. From the CMap analysis six compounds were initially predicted as most prominent to reverse the high-risk non-muscle invasive molecular signature according to their high relative connectivity scores. These included the mTOR inhibitors WYE-354, PP-30 and AZD-8055, the tubulin inhibitor NPI-2358, the PKC activator phorbol-12-myristate-13-acetate (PMA), and the caspase activator PAC-1. WYE-354, PP-30, and AZD-8055 were also retrieved when considering in the CMap analysis features with concordant expression patterns at the proteome and transcriptome level of aggressive NMIBC but also proteomic changes of MIBC (based on our previously published data). Among them WYE-354 was prioritized due to its novelty in the context of BC, its availability as well as the high connectivity scores for reversing both MIBC and aggressive phenotype of NMIBC to early NMIBC.
To establish the drug repurposing pipeline, the impact of WYE-354 was further checked on the tumorigenic properties of BC cell lines representing different stages during disease progression. WYE-354 significantly reduced the proliferation rate of the BC cell lines and impaired their colony forming ability by decreasing the size but not the number of the colonies. No significant impact on apoptosis or necrosis was observed demonstrating the absence of non-specific toxic effects of the inhibitor in the cells. Overall, the in vitro results supported the successful establishment of the drug repurposing pipeline.
To better characterize BC aggressiveness we have also extended the molecular analysis to include also differences observed between NMIBC and MIBC. To this end, different -omic signatures were created and investigated through our established pipeline using tissue proteomics, transcriptomics and omics data retrieved from the literature, but also proteomics and transcriptomics data from BC cell lines. CMap analyses using as input these molecular BC signatures resulted in 23 compounds that may have the potential to reverse the disease signature. Thirteen out of the 23 compounds were not previously evaluated in the context of BC. Three of the 13 compounds demonstrated a prominent impact on cell growth at relatively low concentrations. Additional functional experiments as well as the characterization of the proteomic profile of BC cell lines treated with the three most prominent compounds to unravel the drug response signature are currently ongoing.
Overall, ReDrugBC project resulted in the establishment of a drug repurposing pipeline for the identification of drugs that have the potential of reversing BC; a pipeline that can be further expanded to other cancer types. As part of the pipeline establishment, in vitro assessment of 1 compound was successfully completed. In addition 13 compounds with potential to reverse BC aggressiveness were identified and tested in vitro. Three of them are currently already under evaluation in preclinical animal models. Through the successful implementation of the work planned within ReDrugBC, two research articles and one opinion letter have been already published and additional manuscripts are in preparation. Furthermore, the MSCA-IF Fellow received an invitation to contribute a book chapter describing the developed pipeline within ReDrugBC. The Project results were also presented in international conferences and web-seminars. A website for the project was also launched (
https://www.ReDrugBC.eu(si apre in una nuova finestra)). In parallel to the scientific activities, the MSCA-IF Fellow received an extensive inter-sectorial training that strengthened her skills and supported her career development and independence as a researcher in the industrial setting.
