MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15896/m_MTBLS15896_LC-MS_alternating_hilic_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15896/s_MTBLS15896.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15896/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15896/a_MTBLS15896_LC-MS_alternating_hilic.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15896/FILES/RAW_FILES/20240718_Craig_Priya_lipid.wiffftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15896/FILES/RAW_FILES/20240718_Craig_Priya_lipid.wiff.scanprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15896<p>Hierarchical clustering heatmaps were generated in R using the&nbsp;ComplexHeatmap&nbsp;package</p>MetaboLightsPublicLiquid Chromatography MS - alternating - hilic<p>Chromatographic separation was performed using an Agilent 1290 Infinity LC system equipped with an XBridge Amide column (3.5 µm, 4.6 × 150 mm; Waters) using hydrophilic interaction liquid chromatography (HILIC).</p>DEGS1-mTORC1-driven lipid metabolic reprogramming fuels rhabdomyosarcoma progression and unveils a therapeutic vulnerability.Duke-NUS Medical SchoolPriyadarshini Gopalcellmass spectrometry assay<p> lysate was mixed with 270 µL of ice-cold acetonitrile, followed by vortexing and/or additional sonication to ensure homogeneity. Samples were aliquoted as required for downstream analyses. 25uL of samples &nbsp;were added with&nbsp;25 μL of the premixed lipid internal standard (Sciex lipidyzer platform) extracted in a DCM: Methanol: H20 mixture. Evaporate solvent and reconstitute the extracted lipids in 100 μL of MeOH.&nbsp;</p>Mus musculusHomo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS15896Hong Wen Tang. Duke-NUS Medical School. hongwen.tang@duke-nus.edu.sg.Priyadarshini Gopal. Duke-NUS Medical School. priya.gk@duke-nus.edu.sg.<p>Data were log2-transformed and row-wise z-scored prior to heatmap generation.&nbsp;</p>Cell lineGenetic modificationpriya.gk@duke-nus.edu.sg<p>Cells were cultured to full confluency in standard tissue culture plates. Cells were washed once with 2 mL of ice-cold phosphate-buffered saline (PBS), then incubated with an additional 1 mL of ice-cold PBS and scraped from the plate. After centrifugation at 1,000 x g for 5 minutes at 4 °C, the supernatant was aspirated, and cell pellets were resuspended in 300 µL of ice-cold deionized water containing 0.6% formic acid. Samples were sonicated on ice for 10 minutes to ensure complete lysis.&nbsp;</p>MetabolomicsMus musculustargeted analysisHomo sapiensLipidomicsAB SCIEX QTRAP 5500 SystemcellrhabdomyosarcomaAgilent 1290 Infinity LCDUKE- NUS Metabolomics FacilityMus musculustargeted analysisHomo sapiensLipidomicsAB SCIEX QTRAP 5500 SystemcellrhabdomyosarcomaAgilent 1290 Infinity LCDUKE- NUS Metabolomics Facility<p>Chromatographic separation was achieved using XBridge Amide 3.5 μm, 4.6 × 150 mm column&nbsp;(Waters). Positive ion mode was used to detect SM/CER/DCER/HCER/LCER/TAG/DAG/MAG, while negative ion mode was used to detect PC/PE/PG/PI/PS/FFA.Raw LC–MS data were processed using MultiQuant software (SCIEX). Lipid species were annotated based on parent and daughter fragments and retention time.&nbsp;To check for the quality of data, PCA plots for the samples, blank and internal standard controls&nbsp;were generated using Agilent Mass Profiler Professional (MPP) software.</p>falseDEGS1-mTORC1-driven lipid metabolic reprogramming fuels rhabdomyosarcoma progression and unveils a therapeutic vulnerabilityThese findings establish a mechanistic link between sphingolipid metabolism, mTORC1-mediated regulation of lipophagy, and tumor aggressiveness, highlighting lipid droplet metabolism as a novel and tractable therapeutic target in RMS.2026-10-022026-10-02MTBLS15896