Inhibition of Stearoyl-CoA desaturase 1 reverts BRAF and MEK inhibition-induced selection of cancer stem cells in BRAF-mutated melanoma

Maria Elena Pisanu,Eugenio Tabbì,Mauro Picardo,Paolo A Ascierto,Luigi Fattore,Barbara Bellei,Emilia Migliano,Maria Rosaria Torrisi,Franco Fulciniti,Emanuela Camera,Armando Bartolazzi,Claudia Cippitelli,Žiga Jakopin ,Salvatore Raffa,Gennaro Ciliberto,Marcello Maugeri‐Saccà ,Claudia De Vitis,Sara Bruschini,Daniela Kovacs,Rita Mancini

doi:10.1186/s13046-018-0989-7

Abstract

BackgroundCombination therapy with BRAF and MEK inhibitors significantly improves survival in BRAF mutated melanoma patients but is unable to prevent disease recurrence due to the emergence of drug resistance. Cancer stem cells (CSCs) have been involved in these long-term treatment failures. We previously reported in lung cancer that CSCs maintenance is due to altered lipid metabolism and dependent upon Stearoyl-CoA-desaturase (SCD1)-mediated upregulation of YAP and TAZ. On this ground, we investigated the role of SCD1 in melanoma CSCs.MethodsSCD1 gene expression data of melanoma patients were downloaded from TCGA and correlated with disease progression by bioinformatics analysis and confirmed on patient’s tissues by qRT-PCR and IHC analyses. The effects of combination of BRAF/MEKi and the SCD1 inhibitor MF-438 were monitored by spheroid-forming and proliferation assays on a panel of BRAF-mutated melanoma cell lines grown in 3D and 2D conditions, respectively. SCD1, YAP/TAZ and stemness markers were evaluated in melanoma cells and tissues by qRT-PCR, WB and Immunofluorescence.ResultsWe first observed that SCD1 expression increases during melanoma progression. BRAF-mutated melanoma 3D cultures enriched for CSCs overexpressed SCD1 and were more resistant than 2D differentiated cultures to BRAF and MEK inhibitors. We next showed that exposure of BRAF-mutated melanoma cells to MAPK pathway inhibitors enhanced stemness features by upregulating the expression of YAP/TAZ and downstream genes but surprisingly not SCD1. However, SCD1 pharmacological inhibition was able to downregulate YAP/TAZ and to revert at the same time CSC enrichment and resistance to MAPK inhibitors.ConclusionsOur data underscore the role of SCD1 as prognostic marker in melanoma and promote the use of SCD1 inhibitors in combination with MAPK inhibitors for the control of drug resistance.

Highlights

Combination therapy with BRAF and MEK inhibitors significantly improves survival in BRAF mutated melanoma patients but is unable to prevent disease recurrence due to the emergence of drug resistance
Stearoyl-CoA desaturase 1 (SCD1) is potentially useful for discriminating healthy tissue from melanoma We first sought to determine the diagnostic relevance of SCD1 by analysing RNA levels in melanoma patients
The results indicate an enrichment of SCD1 mRNA expression in 3D spheroids compared with 2D parental cells; k) Western blotting analysis of SCD1 protein in a large panel of melanoma stable and primary cell lines grown in 3D and 2D conditions; l) monounsaturated fatty acids (MUFAs) levels in A375 and M14 cell lines analysed by Gas chromatography-mass spectrometry (GS/MS) in 2D and 3D cultures

Summary

Introduction

Combination therapy with BRAF and MEK inhibitors significantly improves survival in BRAF mutated melanoma patients but is unable to prevent disease recurrence due to the emergence of drug resistance. Given that functional evidence pointed to continued activation of the MAPK pathway as a way cancer cells exploit to overcome the roadblock imposed by BRAF inhibitors, combining BRAF and MEK inhibitors resulted in improved clinical outcomes compared to single agent BRAFdirected therapy [4,5,6]. Despite these therapeutic successes, melanoma cells acquire the ability to withstand combined BRAF and MEK inhibition. Adding further complexity to this picture is the existence, within the same tumour, of distinct cell clones relying on different survival pathways

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