MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15007/m_MTBLS15007_LC-MS_negative_hilic_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15007/a_MTBLS15007_LC-MS_negative_hilic.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15007/s_MTBLS15007.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15007/i_Investigation.txtprimary200OKftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15007<p>Metabolite identification was based on the combination of two criteria: (1) retention time (RT) and (2) the specific MRM ion transition (Q1 -&gt; Q3) , both of which were predetermined using authentic reference standards. The accurate comparison of these parameters against the standards analyzed under identical chromatographic and mass spectrometric conditions ensured the unambiguous identification of each target metabolite in the biological samples.</p>MetaboLightsPublicLiquid Chromatography MS - negative - hilic<p>Chromatographic separation was performed on an ultra-high performance liquid chromatography (UHPLC) system (ExionLC™ AD, AB SCIEX Corp., USA). The separation was carried out on a Waters Atlantis Premier BEH Z-HILIC column (2.1 × 100 mm, 1.7 μm) , which is a hydrophilic interaction liquid chromatography (HILIC) column. The column temperature was maintained at 50°C. The mobile phase consisted of solvent A (15 mM ammonium acetate with 10 µM imino-bis(methylphosphonic acid) in water) and solvent B (15 mM ammonium acetate in acetonitrile). The flow rate was 0.4 mL/min, and the injection volume was 2 µL. The gradient elution program was as follows: 0-5 min, 95% B; 5-8 min, 95-70% B; 8-16 min, 70-40% B; 16-21 min, 40% B; 21-22.1 min, 40-95% B; and 22.1-24 min, 95% B.</p>Hyaluronan catabolism supports the peritoneal disseminated metastasis of cancer through the glucuronic acid pathway.Jie ShiNankai universityOVCAR-8 cellmass spectrometry assay<p>Metabolites were extracted from the samples using a standardized protocol. For a detailed description of the extraction procedure, please refer to the supplementary information (Method.pdf). In brief, the extraction process was optimized for the targeted panel of central carbon metabolism (CCM) metabolites. Following extraction, samples were analyzed in two dilution series (1X and 100X) to ensure all metabolites fell within the dynamic range of the standard curves.</p>Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS15007Jie Shi. Nankai university. 1120220784@mail.nankai.edu.cn.<p>Data processing and quantification were performed using the instrument's software. Metabolite concentrations were determined using a stable isotope-labeled internal standard (IS) method. A 9-point calibration curve was constructed for each metabolite by plotting the peak area ratio of the analyte to its corresponding internal standard against the concentration ratio. Linear regression analysis was performed on the calibration curves, and all correlation coefficients (r) were &gt; 0.99. The limit of quantification (LOQ) for each metabolite was determined using a signal-to-noise ratio of 10:1. The concentrations of metabolites in the original samples were back-calculated from the linear regression equations.</p>Treatment1120220784@mail.nankai.edu.cn<p> Cells (1×107/sample) were quenched in liquid nitrogen and extracted with 80% methanol containing 10 μM norvaline (internal standard). Tests were performed and analyzed on an ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) system (Novogene, Beijing, China). The samples were centrifuged at 1000 rpm for 3 min (4°C) to remove the supernatant. Then homogenized with 250 μL of methanol (80%) which contained mixed internal standards and centrifuged at 15000 rpm for 15 min (4°C) to remove the protein. The supernatant was added to water by well vortexing as the diluted sample. Then 100 μL of them were taken respectively and homogenized with 100 μL of imino-bis (methylphosphonic acid) by well vortexing. After that, centrifuged at 15000 rpm for 15 min. Finally, the supernatant was injected into the LC-MS/MS system for analysis.</p>MetabolomicsMetabolomicsOVCAR-8 cellSCIEX ExionLC ADtargeted analysisOpenMSHomo sapiensmzmineAB SCIEX QTRAP 6500Waters softwareexperimental sampleMetabolomicsOVCAR-8 cellSCIEX ExionLC ADtargeted analysisOpenMSHomo sapiensmzmineAB SCIEX QTRAP 6500Waters softwareexperimental sample<p>Mass spectrometric detection was performed using an AB Sciex QTRAP® 6500+ hybrid triple quadrupole/linear ion trap mass spectrometer equipped with an electrospray ionization (ESI) source. The instrument was operated in negative ionization mode. The scan mode used was multiple reaction monitoring (MRM) . The MRM transitions for each of the 34 target metabolites and 5 internal standards were optimized and are detailed in Table 2-2 of the report. The specific Q1 and Q3 values for each compound were between 87 to 810.3 m/z and 42.8 to 427.9 m/z, respectively. The mass spectrometer parameters were set as follows: IonSpray Voltage, -4500 V; Curtain Gas, 35 psi; Ion Source Temperature, 550°C; and Ion Source Gas 1 and 2, 60 psi.</p>dGDPGDPGTPdATPATPfalseHyaluronan catabolism supports the peritoneal disseminated metastasis of cancer through the glucuronic acid pathwayWhen disseminated into the peritoneal cavity at the very early stage, cancer cells must adapt to the glucose- and oxygen-limited environment in peritoneal fluid, yet the key molecular regulators remain undefined. Here, we reveal that early disseminated OC cells are ingeniously utilize hyaluronic acid as an energy source and a signal molecule to survive in the nutrient-limited peritoneal microenvironment. Through a genome-wide CRISPR/Cas9 knockout screening in an orthotopic ovarian cancer (OC) model, we identified a series of genes involved in hyaluronic acid (HA) catabolism and glucuronic acid (GlcA) metabolism, including the HA receptor LAYN, HA catabolism enzymes (including HYAL1 and HYAL3) and key GlcA metabolic enzymes (such as AKR1A1 and XYLB). By integrating transcriptomic and metabolic analyses in multiple experimental systems, we demonstrated that HA induced the expression of key HA catabolism and GlcA pathway enzymes, which further led to the release of free GlcA from HA degradation. This GlcA is subsequently metabolized through the GlcA pathway, the pentose phosphate pathway (PPP) and glycolysis to support the maintenance and growth of disseminated OC cells. In addition, we found an atypical Rho GTPase RHOU facilitated the LAYN endosomal recycling for efficient HA uptake. Intriguingly, the rewiring of HA catabolism through GlcA pathway was regulated by its classical receptor CD44 and occurred in other peritoneal disseminating cancers such as bladder cancer and pancreatic adenocarcinoma. This signaling-metabolic axis demonstrates the dual role of HA as both a signaling molecule and an energy source supporting cancer cells survival. Importantly, pharmacological inhibition of HYAL1 with garcinol potently suppressed peritoneal disseminated metastasis in xenograft mice and synergized with cisplatin. In this study, we collectively reported a novel HA-induced metabolic reprogramming feature of the early peritoneal disseminated cancer cells, which provides new diagnostic and therapeutic strategies for the cancers prone to the potential dissemination.2026-07-102026-07-10MTBLS15007MTBLC15996MTBLC16284MTBLC28862MTBLC15422MTBLC17552CHEBI:15996CHEBI:16284CHEBI:28862CHEBI:15422CHEBI:17552