Project Funding Details

BACKGROUND: Melanoma incidence is still rising, making it the 6th most prevalent cancer in the UK. Metastatic disease is largely incurable. 15% of those diagnosed with melanoma will succumb to their disease, accounting for approximately 2000 deaths every year in the UK and, on average, 20 years of life lost per death. Notwithstanding recent clinical success through targeting mutant BRAF, new therapeutic avenues must still be explored if advanced melanoma is to be cured. The extensive reprogramming of intracellular signalling, metabolism and gene expression in advanced melanoma cells makes target selection a significant challenge. Screening of biologically active compounds in a whole-organism cancer model represents an alternative, efficient and unbiased method of probing the proteome for therapeutic targets. Increasingly, combining drugs is seen as a way of improving efficacy; an added challenge is to identify safe and effective drug combinations.AIMS & OBJECTIVES: The overall aim of our proposal is to identify pharmacological targets that co-operate with melanoma drivers to induce melanoma and to subsequently validate them as combinatorial therapeutic targets. Our specific objectives are: 1.To identify small molecule hits from libraries of biologically active compounds that in combination with PD184352 or NVPBEZ235 selectively antagonize melanoma onset in a RAS-driven zebrafish transgenic melanoma model we have recently developed.2.To assess the ability of hits to antagonize the proliferation or survival of human melanoma cells in 3D culture models and to inhibit melanoma growth in zebrafish and mouse models. 3.To assess the mode of action of hits through gene knock-down approaches as well as by target identification. METHODS: Libraries of compounds of known biological activity, including licensed drugs, will be screened for Hits that suppress the development of melanoma in V12RAS-expressing zebrafish embryos when combined with either PD184352 (a MEK inihibitor) or NVPBEZ235 (a dual PI3K/MTOR inhibitor). Activity of the combined agents will then be confirmed in 3D culture models of human melanoma cells, as well as in tumours in adult zebrafish and human melanoma xenografts in mouse. The mechanism of action of Hits will be investigated by gene knock-down, but since the targets of Hits may not always be known, we also propose employing Magic tag technology to identify putative targets in these instances.HOW THE RESULTS OF THIS RESEARCH WILL BE USED: Validated therapeutic targets could serve as entry points for future rational drug design projects. Alternatively, the screen may repurpose existing drugs.