MetaboLights

application/xml

ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14024/m_MTBLS14024_LC-MS_alternating_reverse-phase_metabolite_profiling_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14024/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14024/a_MTBLS14024_LC-MS_alternating_reverse-phase_metabolite_profiling.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14024/s_MTBLS14024.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14024Mass spectrometry-based quantitative metabolomics refers to the determination of the concentration of a substance in an unknown sample by comparing the unknown to a set of standard samples of known concentration (i.e., calibration curve). The calibration curve is a plot of how the analytical signal changes with the concentration of the analyte (the substance to be measured). For most analyses a plot of instrument response vs. concentration will show a linear relationship. This yields a model described by the equation y = ax + b, where y is the instrument response e.g., peak height or area, a represents the slope/sensitivity, and b is a constant that describes the background. The analyte concentration (x) of unknown samples may be calculated from this equation.MetaboLightsPublicLiquid Chromatography MS - alternating - reverse-phaseA ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) system (ACQUITY UPLC-Xevo TQ-S, Waters Corp., Milford, MA, USA) was used to quantitate a l l t a r g e t e d metabolite s in this project.The optimized instrument settings are briefly described below. The instrument performance optimization and routine maintenance were performed every week.ACOD1 deficiency promotes dermal papilla cell senescence via mitochondrial dysfunction in androgenetic alopecia.B SZhejiang University of Technologypmol/million cellsmass spectrometry assayEach cell sample that was harvested and stored in an Eppendorf Safelock microcentrifuge tube, was mixed with 10 pre-chilled zirconium oxide beads and 20μL of deionized water. The sample was homogenated for 3 minutes and 150μL of Methanol containing internal standard was added to extract the metabolites. The sample was homogenated for another 3 minutes and then centrifuged at 18000g for 20 minutes. Then the supernatant was transferred to a 96-well plate. The following procedures were performed on a Eppendorf epMotion Workstation(Eppendorf Inc., Humburg, Germany). 20μL of freshly prepared derivative reagents was added to each well. The plate was sealed and the derivatization was carried out at 30° Cfor 60 min. After derivatization, the sample was evaporated for 2h. 330μL of ice-cold 50% methanol solution was added to reconstitute the sample. Then the plate was stored at -20° Cfor 20 minutes and followed by 4000g centrifugation at 4 °C for 30 minutes. 135μL of supernatant was transferred to a new 96-well plate with 10μL internal standards in each well. Serial dilutions of derivatized stock standards were added to the left wells. Finally the plate was sealed for LC-MS analysis.Homo sapienshttps://www.ebi.ac.uk/metabolights/MTBLS14024Weixin Fan. Department of Dermatology, the First Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu, 210029, China. hairmanfwx@163.com.Min Zhao. Department of Dermatology, the First Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu, 210029, China. zhaomin20201122@sina.com.The raw data files generated by UPLC-MS/MS were processed using the TMBQ software (v1.0, Metabo-Profile, Shanghai, China) to perform peak integration, calibration, and quantitation for each metabolite. The platform iMAP (v1.0, Metabo-Profile, Shanghai, China) was used for statistical analyses, including PCA, OPLS-DA, univariate analysis and pathway analysis, et al. Treatment253313593@qq.comCollected cell samples were homogenized with pre-chilled zirconium beads and deionized water.MetabolomicsSenescenceMitochondrial dysfunctionDDX1Dermal papilla cellsACOD1Androgenetic alopeciaSenescenceMitochondrial dysfunctionDDX1Dermal papilla cellsACOD1Androgenetic alopeciaA ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) system (ACQUITY UPLC-Xevo TQ-S, Waters Corp., Milford, MA, USA) was used to quantitate a l l t a r g e t e d metabolite s in this project.The optimized instrument settings are briefly described below. The instrument performance optimization and routine maintenance were performed every week.HMDB0000158HMDB0000714HMDB0000756HMDB0002931HMDB0000679HMDB0000634HMDB0003229HMDB0000716HMDB0000759HMDB0000517HMDB0000719HMDB0000191HMDB0013713HMDB0000190HMDB0001043HMDB0244268HMDB0002250HMDB0000673HMDB0000156HMDB0000232HMDB0002013HMDB0000791HMDB0000230HMDB0000197HMDB0000622HMDB0002925HMDB0000824HMDB0000867HMDB0001999HMDB0000821HMDB0006528HMDB0000904HMDB0013128HMDB0006248HMDB0000064HMDB0000062HMDB0000182HMDB0000222HMDB0000023HMDB0000187HMDB0060038HMDB0000696HMDB0000736HMDB0000812HMDB0000613HMDB0002271HMDB0000092HMDB0012328HMDB0006270HMDB0000172HMDB0000094HMDB0000651HMDB0001388HMDB0000134HMDB0005066HMDB0000452HMDB0000177HMDB0000056HMDB0000011HMDB0000176HMDB0000098HMDB0000721HMDB0000687HMDB0002226HMDB0000201HMDB0000168HMDB0000641HMDB0000725HMDB0000207HMDB0000008HMDB0001976HMDB0002108HMDB0000848HMDB0000529HMDB0000606HMDB0000929HMDB0013302HMDB0003073HMDB0013622HMDB0002183HMDB0003070HMDB0000162HMDB0000161HMDB0006469HMDB0006029HMDB0000621|HMDB0001644HMDB0000123HMDB0000167HMDB0000484HMDB0029765falseACOD1 deficiency promotes dermal papilla cell senescence via mitochondrial dysfunction in androgenetic alopeciaBackground Androgenetic alopecia (AGA), the most prevalent form of hair loss, is driven by the dysfunction of dermal papilla cells (DPCs). Emerging evidence implicates DPC senescence in the pathogenesis of AGA; however, the underlying molecular mechanisms remain incompletely elucidated.Methods We employed a multi-faceted experimental approach, including analyses of human scalp tissues, primary DPCs, a dihydrotestosterone (DHT)-induced AGA mouse model, and immortalized DPCs stimulated with DHT. Cellular senescence was evaluated via expression of senescence markers (p16INK4a, p21, p53) and senescence-associated β-galactosidase staining. Cell proliferation, migration, and apoptosis were also assessed. Mitochondrial function was evaluated using transmission electron microscopy, MitoTracker, MitoSOX, JC-1, and Seahorse assays. RNA sequencing and bioinformatics analyses were performed to identify differentially expressed genes. The interaction between Aconitate Decarboxylase 1 (ACOD1) and DDX1 was verified via co-immunoprecipitation, mass spectrometry, and molecular docking. Metabolomic analysis was performed to profile intracellular metabolic alterations. Functional experiments included ACOD1 knockdown/overexpression and exogenous supplementation with 4-octyl itaconate (4-OI). Hair follicle morphology and hair loss in AGA mice were evaluated following 4-OI treatment.Results Senescence markers were significantly elevated in AGA DPCs, accompanied by increased senescence-associated β-galactosidase staining, reduced cell proliferation and migration, and enhanced apoptosis. DHT-induced mitochondrial dysfunction in DPCs was characterized by increased mitochondrial fragmentation, decreased mitochondrial cristae, superoxide accumulation, reduced membrane potential, and impaired oxidative phosphorylation. RNA sequencing identified ACOD1 as significantly downregulated in DHT-treated DPCs. ACOD1 knockdown induced mitochondrial dysfunction, cellular senescence, and functional impairment in DPCs, while ACOD1 overexpression rescued these DHT-induced phenotypes. Mechanistically, ACOD1 interacted with DDX1 to inhibit its methylation; ACOD1 knockdown enhanced DDX1 methylation, thereby promoting mitochondrial dysfunction and DPC senescence. Metabolomic analysis demonstrated that ACOD1 knockdown significantly reduced itaconate levels. Exogenous 4-OI supplementation reduced DDX1 methylation, ameliorated DHT-induced mitochondrial dysfunction and cellular senescence, promoted cell proliferation and migration, suppressed apoptosis, mitigated hair follicle miniaturization, and alleviated hair loss in AGA mice.Conclusions Our findings reveal a novel mechanism underlying DPC senescence in AGA, wherein ACOD1 deficiency promotes DDX1 methylation, mitochondrial dysfunction, and subsequent DPC senescence. ACOD1 represents a promising therapeutic target for AGA, and 4-OI may have translational potential for the treatment of AGA.2026-05-232026-03-11MTBLS14024CreatineCitrullineCitraconic acidCitric acidCarnitine