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spt10.D/Results/Qual/Version4/default.m/DaMethod/Qual/qualitative.xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS11746/FILES/RAW_FILES/METABOLOMICS/DSIM_12_spt10.D/AcqData/sample_info.xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS11746/FILES/RAW_FILES/DSIM_38_spt10.D/Results/Qual/Version4/default.m/DaMethod/Qual/qualitative.xmlprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS11746<p>Data was acquired in MRM mode, the method development, (MRM transitions and Retention time) was built up by synthetic standards.</p>MetaboLightsPublicGas Chromatography MS - positive<p>In total 1 µL of the derivatized sample was injected by an Agilent 7693 autosampler, in splitless mode into an Agilent 7890A gas chromatograph equipped with a multimode inlet (MMI) and 10 m x 0.18 mm fused silica capillary column with a chemically bonded 0.18 μm DB 5-MS UI stationary phase (J&amp;W Scientific). The injector temperature was 260 °C. The carrier gas flow rate through the column was 1 mL/min, the column temperature was held at 70 °C for 2 min, then increased by 40 °C/min to 320 °C and held there for 2 min.</p>Parallel single-cell metabolic analysis and extracellular vesicle profiling reveal vulnerabilities with prognostic significance in acute myeloid leukemia.University of BolognaDorian ForteExtracellular Vesiclemass spectrometry<p>EV isolated following our protocols were diluted in 80% methanol (1:10), vortex, and then centrifuged for 10 min at 4 °C at 18,000 x g.&nbsp;Analysis of the TCA cycle intermediates was performed by GC-QQQ-MS at Swedish Metabolomics Centre. </p><p><br></p><p>For Gas Chromatography-Mass Spectrometry&nbsp;(GC-MS) analysis an 11-point calibration curve (cis-aconitic acid, a-keto-glutaric acid, citric acid, fumaric acid, glucose, glucose 6-phosphate, isocitric acid, lactic acid, malic acid, shikimic acid, succinic acid, sucrose, urea) spanning from 25-2500 pg/µL was prepared by serial dilutions and spiked with internal standards (Fumaric acid (13C4), L-Malic acid (13C4), D-Glucose (13C6), α-ketoglutaric acid (13C4), Succinic acid (D4), citric acid&nbsp;(D4), Sucrose (13C12) at a final concentration of 350 pg/μL. Supernatants were frozen until use. Up to 200 µL of the extracted sample were transferred to GC vials, and the above-mentioned isotopically labeled internal standards were added. The sample was evaporated to dryness under a stream of nitrogen.</p><p><br></p><p>Derivatization was performed according to Gullberg et al 66. In detail, 10 µL of methoxyamine (15 µg/µL in pyridine) was added to the dry sample that was shaken vigorously for 10 min before left to react in room temperature. After 16 h 20 µL of a mixture of MSTFA and methyl stearate (1050 pg/µL in heptane) (1:1) was added and the sample was vortexed.</p>Homo sapiens<p>Data were processed using MassHunter Qualitative Analysis and Quantitative Analysis (QqQ; Agilent Technologies, Atlanta, GA, USA) and Excel (Microsoft, Redmond, Washington, USA) software. Methyl stearate was used as internal standard for compounds lacking a stable isotope internal standard.</p>diseaserisk factorhttps://www.ebi.ac.uk/metabolights/MTBLS11746dorian.forte2@unibo.it<p>For this study we used extracellular vesicles (EVs) isolated from peripheral blood plasma of acute myeloid leukemia patients and healthy donors. EVs were isolated using size-exclusion chromatography and ultrafiltration. </p>MetabolomicsCitrate Cycle (TCA Cycle) PathwayExtracellular Vesicletargeted metabolitesCitrate Cycle (TCA Cycle) PathwayExtracellular Vesicletargeted metabolites<p>The column effluent is introduced into the electron impact (EI) ion source of an Agilent 7000C QQQ mass spectrometer. The thermal AUX 2 (transfer line) and the ion source temperatures were 250 and 230 °C, respectively. Ions were generated by a 70 eV electron beam at an emission current of 35 µA and analyzed in dMRM-mode.&nbsp;The solvent delay was set to 2 min.</p>lactic acidfumaric acidglucosemalic acidsucroseaconitic acidshikimic acidureaalpha-keto-glutaric acidcitric acidglucose-6-phosphatesuccinic acidisocitric acidfalseParallel single-cell metabolic analysis and extracellular vesicle profiling reveal vulnerabilities with prognostic significance in acute myeloid leukemia (Metabolomic analysis on extracellular vesicles)<p>Acute myeloid leukemia (AML) is an aggressive disease with a high relapse rate. In this study, we map the metabolic profile of CD34+(CD38low/-) AML cells and the extracellular vesicle signatures in circulation from AML patients at diagnosis. CD34+ AML cells display high antioxidant glutathione levels and enhanced mitochondrial functionality, both associated with poor clinical outcomes. Although CD34+ AML cells are highly dependent on glucose oxidation and glycolysis for energy, those from intermediate- and adverse-risk patients reveal increased mitochondrial dependence. Extracellular vesicles from AML are mainly enriched in stem cell markers and express antioxidant GPX3, with their profiles showing potential prognostic value. Extracellular vesicles enhance mitochondrial functionality and dependence on CD34+ AML cells via the glutathione/GPX4 axis. Notably, only extracellular vesicles from adverse-risk patients enhance leukemia cell engraftment in vivo. Here, we show a potential noninvasive approach based on liquid ‘cell-extracellular vesicle’ biopsy toward a redefined metabolic stratification in AML.</p><p><br></p><p><strong>Metabolomic analysis on extracellular vesicles</strong> is reported in the current study <a href='https://www.ebi.ac.uk/metabolights/MTBLS11746' rel='noopener noreferrer' target='_blank'><strong>MTBLS11746</strong></a>.</p><p><strong>Lipidomic analysis on extracellular vesicles</strong> is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS11523' rel='noopener noreferrer' target='_blank'><strong>MTBLS11523</strong></a>.</p>2024-12-042024-11-22MTBLS11746MTBLC22211MTBLC30915MTBLC30769MTBLC18012MTBLC15903MTBLC4170MTBLC30887MTBLC422MTBLC30797MTBLC16119MTBLC15741MTBLC17992MTBLC16199CHEBI:22211CHEBI:30915CHEBI:30769CHEBI:18012CHEBI:15903CHEBI:4170CHEBI:30887CHEBI:422CHEBI:30797CHEBI:16119CHEBI:15741CHEBI:17992CHEBI:16199