MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/m_MTBLS15598_LC-MS_positive_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/s_MTBLS15598.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/a_MTBLS15598_LC-MS_positive_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/256_KO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/94_WT_normalchow.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/75_DKO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/257_KO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/91_WT_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/90_WT_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/259_WT_normalchow.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/202_KO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/254_WT_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/201_WT_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/215_DKO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/208_WT_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/262_KO_normalchow.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/260_WT_normalchow.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/109_DKO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/95_KO_normalchow.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/116_KO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/209_KO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/Blank.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/210_DKO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/115_WT_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/93_KO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/108_DKO_MCD.mzXMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598/FILES/DERIVED_FILES/92_KO_MCD.mzXMLprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15598<p>Lipid species identities were confirmed by MS2 analysis and retention time of authentic isotope-labeled lipid standard mixtures and fatty acids purchased from Avanti and Sigma. For MS/dd-MS2, the properties are: resolution, 17,500; AGC target was 1e5; maximum IT, 50ms; loop count, 15; isolation window, 1.2m/z; stepped CE, ± 20 and 50eV with the following operational parameters: sheath gas flow rate, 40; aux gas flow rate, 10; sweep gas flow rate, 2; spray voltage, + 3.8kV and −3.5kV; spray current, 33μA; capillary temperature, 300°C; s-lens RF level, 50; aux gas heater temperature, 360°C. Data were analyzed using the EI-MAVEN software.</p>MetaboLightsPublicLiquid Chromatography MS - positive - reverse-phase<p>LC separation was achieved on a Poroshell 120 EC-C18 column (2.1 x 150 mm2, 2.7 μm particle size; Agilent) using a gradient of solvent A (90:10 water:methanol with 1 mM of ammonium acetate and 0.2% acetic acid) and solvent B (2:98 methanol:isopropanol with 1 mM of ammonium acetate and 0.2% acetic acid). The flow rate was 150 μL/min. The LC gradient was 0 min, 25% B; 2 min, 25% B; 4 min, 65% B; 16 min, 100% B; 20 min, 100% B; 21 min, 25% B; 27 min, 25% B. The autosampler temperature was 5°C and the injection volume was 3 μL.Please update this protocol description</p>Selective mTOR inhibition via FLCN drives TFE3-dependent VLDL-TG secretion and protects from MAFLD/MASH.Izabelle LeUniversity of California, IrvineblankLivermass spectrometry assay<p>Approximately 20 mg of homegenized liver powder was dissolved in 1 mL of ice-cold isopropanol, and subsequently centrifuged at 16,000g at 4°C for 10 minutes. The resulting supernatant was transferred to a glass vial for LC-MS analysis</p>Mus musculusblankhttps://www.ebi.ac.uk/metabolights/MTBLS15598Zolt Arany. University of Pennsylvania. TRC 11-106, 3400 Civic Ctr Blvd University of Pennsylvania Philadelphia, PA 19104. zarany@pennmedicine.upenn.edu.<p>Data were analyzed using the EI-MAVEN software</p>DietGenotypeimle@uci.edu<p>With the exception of the in vivo DNL assays, all mice were fasted for 4-5 hours before experiment/sacrifice, with fasting starting between 9am and 10:30am. For VLDL-TG secretion assays, blood was collected retro-orbitally with heparin-coated Natelson tubes (Fisherbrand, 02-668-10) at time points 0h and 4h, and via tail snip using heparin-coated microvette tubes (Sarstedt, 16.443.100) at time points 1h and 2h. Livers were quickly dissected after the terminal 4h blood collection timepoint, and rapidly flash-frozen in liquid nitrogen. For in vivo DNL assays and MAFLD/MASH studies, blood was collected retroorbitally immediately before sacrifice. Livers were dissected and rapidly flash-frozen in liquid nitrogen. All blood samples were centrifuged at 10,000 rpm for 7 minutes at 4°C, and plasma was collected. All samples were stored at -80°C until processing.</p>MetabolomicsMus musculusblankuntargeted analysisLiverThermo Scientific Vanquish Flex UHPLC Systemsolvent blankLipidomicstargeted metabolite profilingmTORC1Thermo Scientific Q Exactive Plusmetabolic dysfunction-associated steatohepatitisexperimental sampleMus musculusblankuntargeted analysisThermo Scientific Vanquish Flex UHPLC SystemLiversolvent blankLipidomicstargeted metabolite profilingmTORC1metabolic dysfunction-associated steatohepatitisThermo Scientific Q Exactive Plusexperimental sample<p>Lipid species were detected with a quadrupole orbitrap mass spectrometer (Q Exactive Plus) operating in positive ion mode with electrospray ionization. The following scan settings were used: m/z 350 to 1,700 at 1 Hz, with a 140,000 resolution.</p>falseSelective mTOR inhibition via FLCN drives TFE3-dependent VLDL-TG secretion and protects from MAFLD/MASHHepatic lipid homeostasis is controlled by multiple processes, including VLDL-TG secretion. We have previously shown that the FLCN/TFE3 branch of mTORC1 signaling controls hepatic fatty acid oxidation and de novo lipogenesis to protect mice from MAFLD. We now show that the FLCN/TFE3 pathway also controls VLDL-TG secretion via direct transcriptional activation of a broad program of genes involved in VLDL-TG secretion, including PCYT1A, the rate-limiting step of phosphatidylcholine synthesis. Loss of hepatic FLCN upregulates VLDL-TG secretion and prevents MAFLD, and this protection requires TFE3 and PCYT1A. The data unveil a new pathway that orchestrates multiple steps of VLDL-TG synthesis and secretion. The data also highlight that suppressing the selective FLCN/TFE3 branch of mTORC1 signaling maximizes hepatic lipid clearance by coordinately promoting fatty acid oxidation, suppressing de novo lipogenesis, and boosting VLDL-TG secretion, providing an attractive therapeutic target for the treatment of MAFLD.2026-09-082026-09-08MTBLS15598