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normalizing the original peak area information with the total peak area, the follow-up analysis was performed. Principal component analysis and Spearman correlation analysis were used to judge the repeatability of the samples within group and the quanlity control samples. The identified compounds are searched for classification and pathway information in KEGG, HMDB and lipidmaps databases.According to the grouping information, calculate and compare the difference multiples, T test was used to calculate the difference significance pvalue of each compound. The R language package ropls was used to perform OPLS-DA modeling, and 200 times permutation tests was performed to verify the reliability of the model. The VIP value of the model was calculated using multiple cross-validation. The method of combining the difference multiple, the P value and the VIP value of the OPLS-DA model was adopted to screen the differential metabolites. The screening criteria are FC>1, P value<0.05 and VIP>1. The difference metabolites of KEGG pathway enrichment significance were calculated using hypergeometric distribution test.MetaboLightsPublicLiquid Chromatography MS - negative - reverse phaseLiquid Chromatography MS - positive - reverse phaseThe LC/MS system used for metabolomics analysis consists of a Waters ACQUITY UPLC I-Class PLUS System. The chromatographic column used is an Acquity UPLC CSH C18 column (1.7um 2.1*100mm) purchased from Waters. Positive ion mode: Mobile phase A: 60% acetonitrile in water, 10mM ammonium acetate, 0.1% formic acid; Mobile phase B: 90% isopropanol acetonitrile solution, 10mM ammonium acetate, 0.1% formic acid. Negative ion mode: Mobile phase A: 60% acetonitrile in water, 10mM ammonium acetate, 0.1% formic acid; Mobile phase B: 90% isopropanol acetonitrile solution, 10mM ammonium acetate, 0.1% formic acid. Injection volume 5ul, column temperature 55 degrees CelsiusIdentification of AK4 and RHOC as novel oncogenes addicted by adult T-cell leukemia.benquan liuChina Pharmaceutical UniversityT-cell acute lymphoblastic leukemia celladult T-cell lymphoma cellcell extractsmass spectrometry assayWeigh 50mg sample into EP tube and add 200μL water. Add 480μL extract (methyl tert-butyl ether: methanol = 5:1), add steel beads, grind at 1000rpm for 12min, vortex mix for 30s, and ultrasonicate for 15min (ice water bath). Let stand at -20 °C for 1h. Centrifuge the sample at 4 °C, 13000rpm for 15min, take 380μL supernatant into EP tube, and vacuum dry. Add 200μL solution (dichloromethane: methanol = 1:1) for re-dissolution, vortex for 30s, and ultrasonicate in ice water bath for 10 minutes. Centrifuge the sample at 4 °C, 12000rpm for 15min. Carefully take out 180μL supernatant into the injection bottle, take 10μL of each sample and mix into QC sample for machine detection.Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS11844Xiaorui Zuo.Takashi Matsumoto.Guangyong Ma. 3120070227@stu.cpu.edu.cn.Kosuke Toyoda.Michi Miura.Yang Sikai.Yuan Xiaoyi.Ruoning Zhou.Yi Liang.Yusuke Higuchi.Jie Liu.Benquan Liu.Jun-ichirou Yasunaga.Masao Matsuoka.The raw data collected using MassLynx V4.2 were processed by Progenesis QI software for peak extraction, peak alignment and other data processing operations. Identification was performed based on the Progenesis QI software online database and Biomarker's self-built library, and theoretical fragment identification was performed at the same time, the deviation of the parent ion mass number was within 100ppm and the deviation of the fragment ion mass number was within 50ppm.Disease3120070227@stu.cpu.edu.cnATL cell lines: MT-4, Hut102, ED and TL-Om1, T-ALL cell lines: Jurkat, CEM-T4, MOLT4 and CTCL cell line HuT 78, are grown in RPMI-1640 supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (100 μg/mL). When the number of cultured cells is >1.0 x 10^7, collect the cells and centrifuge at 1200rpm and 4 °C for 5min. After centrifugation, remove the supernatant, precipitate the cells, count them, and the number of cells in each sample should be at least 1 x 10^7. Then add pre-cooled PBS to the cells, mix well and transfer to a 1.5ml centrifuge tube. Centrifuge at 900rpm and 4 °C for 3min, and remove the supernatant. Repeat the washing of cells 3 times. Then soak in liquid nitrogen for >3min to quench. Store in a -80 °C refrigerator.MetabolomicsMIR455HTLV-1AK4RHOCMIR455HTLV-1AK4RHOCWaters Xevo G2-XS QTof high-resolution mass spectrometer: the MSe mode under the control of the acquisition software (MassLynx V4.2, Waters) performs primary and secondary mass spectrometry data acquisition. In each data acquisition cycle, dual-channel data acquisition can be performed simultaneously for low collision energy and high collision energy. Low collision energy 2V, high collision energy range 10~40V, scanning frequency 0.2 seconds per mass spectrum. ESI ion source parameters are as follows: capillary voltage: 2500V (positive ion mode) or -2000V (negative ion mode); cone voltage: 30V; ion source temperature: 120℃; desolvation gas temperature 550℃; backflush gas flow rate: 50L/h; desolvation gas flow rate: 900L/h. The mass-nucleus ratio (m/z) collection range is 50-2000.falseIdentification of AK4 and RHOC as novel oncogenes addicted by adult T-cell leukemiaAdult T-cell leukemia (ATL) is a highly aggressive T-cell malignancy characterized by human T-cell leukemia virus type 1 (HTLV-1) infection. ATL has a very poor prognosis and lacks satisfactory treatments; therefore, it is critical to identify novel targets in ATL cells in order to develop effective targeted therapeutics. Here we report the identification of two novel oncogenes, AK4 and RHOC, as target genes of miR-455-3p, a tumor suppressive microRNA in ATL patients. Importantly, AK4 and RHOC are highly expressed in ATL and exhibit oncogenic potentials in vitro and in vivo. Interestingly, transcriptome and metabolome analyses reveal a functional overlap of AK4 and RHOC, including activating oncogenic pathways such as Myc targets and deregulating lipid metabolism such as enhancing the production of sphingomyelin, a tumor-promoting lipid. In particular, compared to other types of T-cell malignancy such as T-ALL and CTCL, ATL is sensitive to sphingomyelin inhibition and AK4 or RHOC depletion. Altogether, we report a distinct dependency of ATL on newly characterized oncogenes AK4 and RHOC and an oncometabolite sphingomyelin, which together represent novel targetable vulnerabilities of ATL that could be exploited for developing effective therapeutics.2025-02-032024-12-03MTBLS11844