{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST004042","ANALYSIS_ID":"AN006681","VERSION":"1","CREATED_ON":"08-04-2025"},
"PROJECT":{"PROJECT_TITLE":"Untargeted metabolomics analysis on mouse hepatocytes","PROJECT_SUMMARY":"To determine whether specific metabolites are associated with the induction of P2-HNF4α in liver disease and hepatocellular carcinoma (HCC), we performed untargeted metabolomics on hepatocytes isolated from wild-type mouse liver, which predominantly express the tumor-suppressive P1-HNF4α isoform. Using lentiviral transduction, we overexpressed the P2-HNF4α isoform and conducted metabolite profiling on cell lysates from both control and P2-HNF4α-overexpressing cells. Data analysis was performed using MetaboAnalyst (v5.0), resulting in a list of approximately 120 matched compounds representing significantly predicted metabolites. This study aims to uncover metabolic signatures associated with P2-HNF4α expression, offering insights into its potential role in liver pathogenesis and tumor progression.","INSTITUTE":"University of Texas Health Science Center at Houston","DEPARTMENT":"IMM","LABORATORY":"Mahan Lab","LAST_NAME":"Fekry","FIRST_NAME":"Baharan","ADDRESS":"1825 Pressler St, Houston TX 77030. USA,","EMAIL":"Baharan.Fekry@uth.tmc.edu","PHONE":"7135002487","DOI":"http://dx.doi.org/10.21228/M83G11"},
"STUDY":{"STUDY_TITLE":"Untargeted Metabolomics Identifies Oncometabolites Induced by P2-HNF4α Overexpression in Hepatocytes","STUDY_SUMMARY":"This study explores the metabolic impact of P2-HNF4α overexpression in primary mouse hepatocytes. Using untargeted metabolomics, we compared metabolite profiles between control and P2-HNF4α-overexpressing cells to identify metabolic pathways potentially altered in liver disease and HCC.","INSTITUTE":"University of Tennessee Health Science Center","DEPARTMENT":"IMM","LABORATORY":"Mahan Lab","LAST_NAME":"Fekry","FIRST_NAME":"Baharan","ADDRESS":"1825 Pressler St, Houston TX 77030. USA.","EMAIL":"Baharan.Fekry@uth.tmc.edu","PHONE":"18434693199","SUBMIT_DATE":"2025-07-03"},
"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"HEP_6FP1",
"Factors":{"Sample source":"wildtype","Treatment":"GFP"}
},
{
"Subject ID":"-",
"Sample ID":"HEP_6FP2",
"Factors":{"Sample source":"wildtype","Treatment":"GFP"}
},
{
"Subject ID":"-",
"Sample ID":"HEP_6FP3",
"Factors":{"Sample source":"wildtype","Treatment":"GFP"}
},
{
"Subject ID":"-",
"Sample ID":"HEP_A8_1",
"Factors":{"Sample source":"wildtype","Treatment":"P2_HNF4a"}
},
{
"Subject ID":"-",
"Sample ID":"HEP_A8_2",
"Factors":{"Sample source":"wildtype","Treatment":"P2_HNF4a"}
},
{
"Subject ID":"-",
"Sample ID":"HEP_A8_3",
"Factors":{"Sample source":"wildtype","Treatment":"P2_HNF4a"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Hepatocytes were isolated from mouse livers and transduced in culture with either GFP or P2-HNF4α. Following treatment, cells were collected and subjected to untargeted metabolomics analysis.","SAMPLE_TYPE":"Cultured cells"},
"TREATMENT":{"TREATMENT_SUMMARY":"Primary hepatocytes were isolated from mouse livers and cultured in vitro. Cells were transduced with either control GFP or P2-HNF4α-expressing lentivirus. Following 48 hours of transduction, cells were harvested and processed for untargeted metabolomics analysis."},
"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Samples were diluted in a solution containing 25% acetonitrile, 75% water, and 0.1% formic acid prior to LC-MS analysis. The diluted samples were injected into an Agilent 1200 Series HPLC system coupled to an Agilent 6538 UHD Accurate-Mass Q-TOF mass spectrometer. Chromatographic separation was performed using an Agilent ZORBAX Extend-C18 Rapid Resolution HT column (2.1 × 50 mm, 1.8 µm) under a 20-minute gradient of 25% to 90% Buffer B (80% acetonitrile, 20% water, 0.1% formic acid) at a flow rate of 0.2 mL/min."},
"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Liquid chromatography was performed using an Agilent 1200 Series HPLC system equipped with an Agilent ZORBAX Extend-C18 Rapid Resolution HT column (2.1 × 50 mm, 1.8 µm). The mobile phases consisted of Buffer A (2% acetonitrile, 98% water, 0.1% formic acid) and Buffer B (80% acetonitrile, 20% water, 0.1% formic acid). A linear gradient from 25% to 90% Buffer B was applied over 20 minutes at a flow rate of 0.2 mL/min to achieve metabolite separation prior to MS analysis.","INSTRUMENT_NAME":"Agilent 1200 Series HPLC coupled to Agilent 6538 UHD Accurate-Mass Q-TOF","COLUMN_NAME":"Agilent ZORBAX Extend-C18 Rapid Resolution HT (2.1 × 50 mm, 1.8 µm)","COLUMN_TEMPERATURE":"21°C","FLOW_GRADIENT":"25% to 90% Solvent B over 20 minutes","FLOW_RATE":"0.2 mL/min","SOLVENT_A":"2% Acetonitrile/98% Water; 0.1% formic acid","SOLVENT_B":"80% Acetonitrile/20% Water; 0.1% formic acid","CHROMATOGRAPHY_TYPE":"Reversed phase"},
"ANALYSIS":{"ANALYSIS_TYPE":"MS"},
"MS":{"INSTRUMENT_NAME":"Agilent 6538 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","MS_COMMENTS":"MS1 spectra were acquired over a mass range of 100–2000 m/z using electrospray ionization (ESI) in positive mode. Acquisition was performed under the following conditions: capillary voltage of 3.5 kV, drying gas flow rate of 5 L/min, nebulizer pressure of 5 psi, and source temperature of 300°C.","ION_MODE":"POSITIVE"},
"MS_METABOLITE_DATA":{
"Units":"LC/MS Peak intensity",
"Data":[{"Metabolite":"11_12_15-THETA","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0007","HEP_A8_1":"0.0004","HEP_A8_2":"","HEP_A8_3":"0.0003"},{"Metabolite":"11_14_15-THETA","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0008","HEP_A8_1":"0.0004","HEP_A8_2":"","HEP_A8_3":"0.0003"},{"Metabolite":"11b_17a_21-Trihydroxypreg-nenolone","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0011","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"11b_21-Dihydroxy-3_20-oxo-5b-pregnan-18-al","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0013","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"11-cis-Retinaldehyde","HEP_6FP1":"","HEP_6FP2":"0.0003","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"11-cis-Retinol","HEP_6FP1":"0.0010","HEP_6FP2":"","HEP_6FP3":"0.0003","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"11H-14_15-EETA","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0014","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"1_2-Dihydronaphthalene-1_2-diol","HEP_6FP1":"","HEP_6FP2":"0.0004","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"12(R)-HPETE","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0006","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"12(S)-HPETE","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0003","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"15H-11_12-EETA","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0007","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"15(S)-HPETE","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0000","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"16a-Hydroxyandrost-4-ene-3_17-dione","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0009","HEP_A8_1":"","HEP_A8_2":"0.0010","HEP_A8_3":""},{"Metabolite":"16b-Hydroxyestrone","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0004","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"17alpha_21-Dihydroxypregnenolone","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0002","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"17-beta-Estradiol-3-glucuronide","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0007","HEP_A8_1":"","HEP_A8_2":"0.0005","HEP_A8_3":"0.0004"},{"Metabolite":"17-Hydroxyprogesterone","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"0.0010","HEP_A8_3":""},{"Metabolite":"19-Hydroxyandrost-4-ene-3_17-dione","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0004","HEP_A8_1":"","HEP_A8_2":"0.0010","HEP_A8_3":""},{"Metabolite":"19-Oxoandrost-4-ene-3_17-dione","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0006","HEP_A8_1":"","HEP_A8_2":"0.0005","HEP_A8_3":""},{"Metabolite":"19-Oxotestosterone","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0006","HEP_A8_1":"","HEP_A8_2":"0.0010","HEP_A8_3":""},{"Metabolite":"1a_11b-Dihydro-4_9-dimethylbenz[a]anthra[3_4-b]oxirene","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"0.0014","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"(1aalpha_2beta_3alpha_11calpha)-1a_2_3_11c-Tetrahydro-6_11-dimethylbenzo[6_7]phenanthro[3_4-b]oxirene-2_3-diol","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"","HEP_A8_1":"0.0013","HEP_A8_2":"0.0002","HEP_A8_3":""},{"Metabolite":"1-Methylxanthine","HEP_6FP1":"0.0002","HEP_6FP2":"","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"1-Naphthol","HEP_6FP1":"","HEP_6FP2":"0.0004","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"0.0006","HEP_A8_3":""},{"Metabolite":"1-Nitronaphthalene-5_6-oxide","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"0.0002","HEP_A8_3":""},{"Metabolite":"1-Nitronaphthalene-7_8-oxide","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"0.0005","HEP_A8_3":""},{"Metabolite":"1-Pyrroline-4-hydroxy-2-carboxylate","HEP_6FP1":"","HEP_6FP2":"","HEP_6FP3":"","HEP_A8_1":"0.0001","HEP_A8_2":"","HEP_A8_3":"0.0004"},{"Metabolite":"20a-Dihydroprogesterone","HEP_6FP1":"","HEP_6FP2":"0.0007","HEP_6FP3":"","HEP_A8_1":"","HEP_A8_2":"","HEP_A8_3":""},{"Metabolite":"20-Hydroxy-leukotriene 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