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Modulation of lipid metabolism is a well-established cancer hallmark, and SCD1 has been recognized as a key enzyme in promoting cancer cell growth, including in glioblastoma (GBM), the deadliest brain tumor and a paradigm of cancer resistance. The central goal of this work was to identify, by MS, the phospholipidome alterations resulting from the silencing of SCD1 in human GBM cells, in order to implement an innovative therapy to fight GBM cell resistance. With this purpose, RNAi technology was employed, and low serum-containing medium was used to mimic nutrient deficiency conditions, at which SCD1 is overexpressed. Besides the expected increase in the saturated to unsaturated fatty acid ratio in SCD1 silenced-GBM cells, a striking increase in polyunsaturated chains, particularly in phosphatidylethanolamine and cardiolipin species, was noticed and tentatively correlated with an increase in autophagy (evidenced by the increase in LC3BII/I ratio). The contribution of autophagy to mitigate the impact of SCD1 silencing on GBM cell viability and growth, whose modest inhibition could be correlated with the maintenance of energetically associated mitochondria, was evidenced by using autophagy inhibitors. In conclusion, SCD1 silencing could constitute an important tool to halt GBM resistance to the available treatments, especially when coupled with a mitochondria disrupter chemotherapeutic.

The occurrence of protein synthesis errors (mistranslation) above the typical mean mistranslation level of 10−4 is mostly deleterious to yeast, zebrafish and mammal cells. Previous yeast studies have shown that mistranslation affects fitness and deregulates genes related to lipid metabolism, but there is no experimental proof that such errors alter yeast lipid profiles. We engineered yeast strains to misincorporate serine at alanine and glycine sites on a global scale and evaluated the putative effects on the lipidome. Lipids from whole cells were extracted and analysed by thin layer chromatography (TLC), liquid chromatography-mass spectrometry(LC-MS) and gas chromatography (GC). Oxidative damage, fatty acid desaturation and membrane fluidity changes were screened to identify putative alterations in lipid profiles in both logarithmic (fermentative) and post-diauxic shift (respiratory) phases. There were alterations in several lipid classes, namely lyso-phosphatidylcholine, phosphatidic acid, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and triglyceride, and in the fatty acid profiles, namely C16:1, C16:0, C18:1 and C18:0. Overall, the relative content of lipid species with saturated FA increased in detriment of those with unsaturated fatty acids. The expression of the OLE1 mRNA was deregulated, but phospholipid fluidity changes were not observed. These data expand current knowledge of mistranslation biology and highlight its putative roles in human diseases. This work was supported by FCT(Fundação para a Ciência e a Tecnologia), the European Union, QREN, POPH, FEDER and COMPETE [FCT-ANR/IMI-MIC/0041/2012; PTDC/IMI-MIC/5350/2014; PTDC/BEX-BCM/2121/2014] through the iBiMED (UID/BIM/04501/2013) and QOPNA research units [PTDC/QUI-BIQ/104968/2008; UID QUI/UI0062/2013, FCOMP-01-0124- FEDER-037296], and RNEM [LISBOA-01-0145-FEDER-402-022125 that concerns the Portuguese Mass Spectrometry Network), REDE/1504/REM/2005]. This work was also financed by the European Regional Development Fund (ERDF), through the Centro 2020 Regional Operational Programme under project CENTRO-01-0145-FEDER-000008: BrainHealth 2020, and through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT - Fundação para a Ciência e a Tecnologia, I.P., under projects POCI-01-0145-FEDER-016390:CANCEL STEM and POCI-01-0145-FEDER-007440. Ana R. D. Araújo [SFRH/BD/78835/2011], Catarina Morais [SFRH/BD/79077/2011], Elisabete Maciel [SFRH/BPD/104165/2014], Helena Oliveira [SFRH/BPD/111736/2015], and Tânia Melo [BPD/UI51/5388/2017] were supported by FCT grants. Clara Pereira was supported by Fundo Social Europeu and Programa Operacional Potencial Humano through FCT investigator grant IF/00889/2015.