{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002400","ANALYSIS_ID":"AN003908","VERSION":"1","CREATED_ON":"12-16-2022"},

"PROJECT":{"PROJECT_TITLE":"Alcohol dehydrogenase 1B is crucial for adipocyte homeostasis.","PROJECT_SUMMARY":"Background. Alcohol dehydrogenase (ADH1B), encoded by the ADH1B gene, is a cytosolic enzyme mainly known for its role in ethanol catabolism in the liver. A few studies have paved the way to show an equally important role of ADH1B in adipocytes. This study aimed to better identify the cellular mechanisms and signaling pathways involving ADH1B in adipose tissue and to determine if ADH1B variants might contribute to adipose tissue dysfunction. Results. We showed that CRISPR-Cas9-mediated ADH1B knockout (KO) in human adipose stem cells (ASC) abolished adipocyte differentiation and decreased insulin response. This was accompanied by oxidative stress, altered mitochondrial functions, and cellular senescence. Lipidomic analysis revealed that ADH1B deficiency results in a major remodeling of lipid composition in ASC. An ADH1B homozygous loss-of-function variant was also identified in a patient presenting with a lipodystrophic and insulin resistant syndrome associated with major liver dysfunction, leading to early death. Discussion. This translational study underlines the crucial role of ADH1B in adipose tissue. It unveils cellular mechanisms accounting for its key role in adipogenesis, and adipocyte homeostasis. This study also identifies ADH1B as a candidate gene in monogenic forms of lipodystrophic and insulin resistant syndromes.","INSTITUTE":"INSERM","LAST_NAME":"Gautheron","FIRST_NAME":"Jérémie","ADDRESS":"27 rue Chaligny, 75012 Paris France","EMAIL":"jeremie.gautheron@gmail.com","PHONE":"+33623398373","DOI":"http://dx.doi.org/10.21228/M8VM64"},

"STUDY":{"STUDY_TITLE":"Alcohol dehydrogenase 1B is crucial for adipocyte homeostasis","STUDY_SUMMARY":"Background. Alcohol dehydrogenase (ADH1B), encoded by the ADH1B gene, is a cytosolic enzyme mainly known for its role in ethanol catabolism in the liver. A few studies have paved the way to show an equally important role of ADH1B in adipocytes. This study aimed to better identify the cellular mechanisms and signaling pathways involving ADH1B in adipose tissue and to determine if ADH1B variants might contribute to adipose tissue dysfunction. Results. We showed that CRISPR-Cas9-mediated ADH1B knockout (KO) in human adipose stem cells (ASC) abolished adipocyte differentiation and decreased insulin response. This was accompanied by oxidative stress, altered mitochondrial functions, and cellular senescence. Lipidomic analysis revealed that ADH1B deficiency results in a major remodeling of lipid composition in ASC. An ADH1B homozygous loss-of-function variant was also identified in a patient presenting with a lipodystrophic and insulin resistant syndrome associated with major liver dysfunction, leading to early death. Discussion. This translational study underlines the crucial role of ADH1B in adipose tissue. It unveils cellular mechanisms accounting for its key role in adipogenesis, and adipocyte homeostasis. This study also identifies ADH1B as a candidate gene in monogenic forms of lipodystrophic and insulin resistant syndromes.","INSTITUTE":"INSERM","LAST_NAME":"Gautheron","FIRST_NAME":"Jérémie","ADDRESS":"27 rue Chaligny","EMAIL":"jeremie.gautheron@gmail.com","PHONE":"+33623398373","SUBMIT_DATE":"2022-12-13"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Homo sapiens"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"ADH1B_KO_01_LIP329",
"Sample ID":"ADH1B_KO_01_LIP329",
"Factors":{"Genotype":"ADH1B_KO"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_KO_01","RAW_FILE_NAME":"LIP329_C18_10x_KO_01","RAW_FILE_NAME":"LIP329_long_1x_KO_01","RAW_FILE_NAME":"LIP329_short_1x_KO_01","RAW_FILE_NAME":"LIP329_short_20x_KO_01"}
},
{
"Subject ID":"ADH1B_KO_02_LIP329",
"Sample ID":"ADH1B_KO_02_LIP329",
"Factors":{"Genotype":"ADH1B_KO"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_KO_02","RAW_FILE_NAME":"LIP329_C18_10x_KO_02","RAW_FILE_NAME":"LIP329_long_1x_KO_02","RAW_FILE_NAME":"LIP329_short_1x_KO_02","RAW_FILE_NAME":"LIP329_short_20x_KO_02"}
},
{
"Subject ID":"ADH1B_KO_03_LIP329",
"Sample ID":"ADH1B_KO_03_LIP329",
"Factors":{"Genotype":"ADH1B_KO"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_KO_03","RAW_FILE_NAME":"LIP329_C18_10x_KO_03","RAW_FILE_NAME":"LIP329_long_1x_KO_03","RAW_FILE_NAME":"LIP329_short_1x_KO_03","RAW_FILE_NAME":"LIP329_short_20x_KO_03"}
},
{
"Subject ID":"ADH1B_KO_04_LIP329",
"Sample ID":"ADH1B_KO_04_LIP329",
"Factors":{"Genotype":"ADH1B_KO"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_KO_04","RAW_FILE_NAME":"LIP329_C18_10x_KO_04","RAW_FILE_NAME":"LIP329_long_1x_KO_04","RAW_FILE_NAME":"LIP329_short_1x_KO_04","RAW_FILE_NAME":"LIP329_short_20x_KO_04"}
},
{
"Subject ID":"ADH1B_KO_05_LIP329",
"Sample ID":"ADH1B_KO_05_LIP329",
"Factors":{"Genotype":"ADH1B_KO"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_KO_05","RAW_FILE_NAME":"LIP329_C18_10x_KO_05","RAW_FILE_NAME":"LIP329_long_1x_KO_05","RAW_FILE_NAME":"LIP329_short_1x_KO_05","RAW_FILE_NAME":"LIP329_short_20x_KO_05"}
},
{
"Subject ID":"CTL_01_LIP329",
"Sample ID":"CTL_01_LIP329",
"Factors":{"Genotype":"CTL"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_CTL_01","RAW_FILE_NAME":"LIP329_C18_10x_CTL_01","RAW_FILE_NAME":"LIP329_long_1x_CTL_01","RAW_FILE_NAME":"LIP329_short_1x_CTL_01","RAW_FILE_NAME":"LIP329_short_20x_CTL_01"}
},
{
"Subject ID":"CTL_02_LIP329",
"Sample ID":"CTL_02_LIP329",
"Factors":{"Genotype":"CTL"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_CTL_02","RAW_FILE_NAME":"LIP329_C18_10x_CTL_02","RAW_FILE_NAME":"LIP329_long_1x_CTL_02","RAW_FILE_NAME":"LIP329_short_1x_CTL_02","RAW_FILE_NAME":"LIP329_short_20x_CTL_02"}
},
{
"Subject ID":"CTL_03_LIP329",
"Sample ID":"CTL_03_LIP329",
"Factors":{"Genotype":"CTL"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_CTL_03","RAW_FILE_NAME":"LIP329_C18_10x_CTL_03","RAW_FILE_NAME":"LIP329_long_1x_CTL_03","RAW_FILE_NAME":"LIP329_short_1x_CTL_03","RAW_FILE_NAME":"LIP329_short_20x_CTL_03"}
},
{
"Subject ID":"CTL_04_LIP329",
"Sample ID":"CTL_04_LIP329",
"Factors":{"Genotype":"CTL"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_CTL_04","RAW_FILE_NAME":"LIP329_C18_10x_CTL_04","RAW_FILE_NAME":"LIP329_long_1x_CTL_04","RAW_FILE_NAME":"LIP329_short_1x_CTL_04","RAW_FILE_NAME":"LIP329_short_20x_CTL_04"}
},
{
"Subject ID":"CTL_05_LIP329",
"Sample ID":"CTL_05_LIP329",
"Factors":{"Genotype":"CTL"},
"Additional sample data":{"Batch":"1","RAW_FILE_NAME":"LIP329_C18_1x_CTL_05","RAW_FILE_NAME":"LIP329_C18_10x_CTL_05","RAW_FILE_NAME":"LIP329_long_1x_CTL_05","RAW_FILE_NAME":"LIP329_short_1x_CTL_05","RAW_FILE_NAME":"LIP329_short_20x_CTL_05"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"ASC were isolated from surgical samples of subcutaneous abdominal adipose tissue from a 25-year-old healthy woman with a normal body mass index (BMI). Adipose tissue was enzymatically digested with collagenase B (0.2%).","SAMPLE_TYPE":"Adipose tissue"},

"TREATMENT":{"TREATMENT_SUMMARY":"After centrifugation, stromal vascular fraction was filtered, rinsed, plated and cultured in α-MEM with 10% Fetal Calf Serum (FCS), 1% GlutaMAX (#35050061, Thermo Fisher Scientific), 1% Penicillin/streptomycin (PS - 10,000 UI/mL), 1% HEPES and Fibroblast Growth Factor-2 (FGF-2 -145 nmol/L). After 24 h, only ASC adhered to plastic surfaces, while other cells were removed after culture medium replacement. ASC were maintained in an undifferentiated state in α-MEM supplemented with 10 % newborn calf serum (#CA-1151500; Biosera, MI, USA), 1% GlutaMAX, HEPES and P/S, and FGF-2 (145 nmol/L). Adipocyte differentiation was induced by treating 2-day post-confluent cultures with high-glucose (25 mmol/L) DMEM supplemented with 10 % FCS, 1 % PS, 1 µmol/L dexamethasone (#D4902; Sigma-Aldrich, MI, USA), 1 µM rosiglitazone (#D4902; Sigma-Aldrich), 250 µM 3-isobutyl-1-methyl xanthine (IBMX) (#I7018; Sigma-Aldrich) and 0.17 µmol/L insulin (#I0516; Sigma-Aldrich) for ten days. The medium was then replaced with high-glucose DMEM supplemented with 10% FCS, 1 % PS, 1 µmol/L rosiglitazone and 0.17 µM insulin, and changed to fresh medium every 2 days until the 20th day."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Lipid extraction. Lipids were extracted from ASC cells according to a modified Folch method. Cell pellets were resuspended in 100µl methanol and supplemented with deuterated internal standards. Lipids were extracted with 1.5 mL chloroform and 650 µL methanol, sonicated for 15 min. Phase separation was triggered by addition of 450 µL of ammonium carbonate (250 mM). Lower organic phase was dried and resuspended in 200 µL of liquid chromatography – mass spectrometry (LC-MS) solvent."},

"CHROMATOGRAPHY":{"INSTRUMENT_NAME":"Shimadzu 20AD","COLUMN_NAME":"Phenomenex Kinetex HILIC (150 x 3mm,2.6um)","COLUMN_TEMPERATURE":"45","FLOW_GRADIENT":"-","FLOW_RATE":"300ul/min","SOLVENT_A":"100% water; 0.2% acetic acid; 30mM ammonium acetate","SOLVENT_B":"100% acetonitrile; 0.2% acetic acid","CHROMATOGRAPHY_TYPE":"HILIC"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 4000 QTrap","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","MS_COMMENTS":"MRM acquisition of narrow chromatographic peaks of low abundant phospho- and sphingolipid classes: cer, PG, PI, PE, PE-P This acquisition is referred to as short_1x","ION_MODE":"POSITIVE"},

"MS_METABOLITE_DATA":{
"Units":"mol% of total lipids",

"Data":[{"Metabolite":"Cer(d18:0/14:0)","ADH1B_KO_01_LIP329":"","ADH1B_KO_02_LIP329":"","ADH1B_KO_03_LIP329":"","ADH1B_KO_04_LIP329":"","ADH1B_KO_05_LIP329":"","CTL_01_LIP329":"0.0007","CTL_02_LIP329":"0.0010","CTL_03_LIP329":"0.0011","CTL_04_LIP329":"0.0008","CTL_05_LIP329":"0.0010"},{"Metabolite":"Cer(d18:0/16:0)","ADH1B_KO_01_LIP329":"0.0094","ADH1B_KO_02_LIP329":"0.0082","ADH1B_KO_03_LIP329":"0.0116","ADH1B_KO_04_LIP329":"0.0123","ADH1B_KO_05_LIP329":"0.0141","CTL_01_LIP329":"0.0233","CTL_02_LIP329":"0.0261","CTL_03_LIP329":"0.0226","CTL_04_LIP329":"0.0262","CTL_05_LIP329":"0.0260"},{"Metabolite":"Cer(d18:0/18:0)","ADH1B_KO_01_LIP329":"0.0016","ADH1B_KO_02_LIP329":"0.0018","ADH1B_KO_03_LIP329":"0.0024","ADH1B_KO_04_LIP329":"0.0015","ADH1B_KO_05_LIP329":"0.0018","CTL_01_LIP329":"0.0028","CTL_02_LIP329":"0.0030","CTL_03_LIP329":"0.0030","CTL_04_LIP329":"0.0033","CTL_05_LIP329":"0.0029"},{"Metabolite":"Cer(d18:0/18:1)","ADH1B_KO_01_LIP329":"","ADH1B_KO_02_LIP329":"","ADH1B_KO_03_LIP329":"0.0019","ADH1B_KO_04_LIP329":"0.0011","ADH1B_KO_05_LIP329":"","CTL_01_LIP329":"0.0011","CTL_02_LIP329":"0.0009","CTL_03_LIP329":"0.0011","CTL_04_LIP329":"0.0011","CTL_05_LIP329":"0.0010"},{"Metabolite":"Cer(d18:0/20:0)","ADH1B_KO_01_LIP329":"","ADH1B_KO_02_LIP329":"","ADH1B_KO_03_LIP329":"0.0025","ADH1B_KO_04_LIP329":"0.0017","ADH1B_KO_05_LIP329":"0.0026","CTL_01_LIP329":"0.0030","CTL_02_LIP329":"0.0045","CTL_03_LIP329":"0.0037","CTL_04_LIP329":"0.0036","CTL_05_LIP329":"0.0030"},{"Metabolite":"Cer(d18:0/22:0)","ADH1B_KO_01_LIP329":"0.0032","ADH1B_KO_02_LIP329":"0.0034","ADH1B_KO_03_LIP329":"0.0044","ADH1B_KO_04_LIP329":"0.0042","ADH1B_KO_05_LIP329":"0.0056","CTL_01_LIP329":"0.0089","CTL_02_LIP329":"0.0091","CTL_03_LIP329":"0.0073","CTL_04_LIP329":"0.0072","CTL_05_LIP329":"0.0067"},{"Metabolite":"Cer(d18:0/23:0)","ADH1B_KO_01_LIP329":"","ADH1B_KO_02_LIP329":"","ADH1B_KO_03_LIP329":"0.0007","ADH1B_KO_04_LIP329":"0.0012","ADH1B_KO_05_LIP329":"0.0013","CTL_01_LIP329":"0.0012","CTL_02_LIP329":"0.0011","CTL_03_LIP329":"0.0018","CTL_04_LIP329":"0.0014","CTL_05_LIP329":"0.0015"},{"Metabolite":"Cer(d18:0/24:0)","ADH1B_KO_01_LIP329":"0.0027","ADH1B_KO_02_LIP329":"0.0019","ADH1B_KO_03_LIP329":"0.0037","ADH1B_KO_04_LIP329":"0.0028","ADH1B_KO_05_LIP329":"0.0042","CTL_01_LIP329":"0.0067","CTL_02_LIP329":"0.0056","CTL_03_LIP329":"0.0052","CTL_04_LIP329":"0.0059","CTL_05_LIP329":"0.0045"},{"Metabolite":"Cer(d18:0/24:1)","ADH1B_KO_01_LIP329":"0.0038","ADH1B_KO_02_LIP329":"0.0037","ADH1B_KO_03_LIP329":"0.0041","ADH1B_KO_04_LIP329":"0.0062","ADH1B_KO_05_LIP329":"0.0056","CTL_01_LIP329":"0.0115","CTL_02_LIP329":"0.0132","CTL_03_LIP329":"0.0124","CTL_04_LIP329":"0.0135","CTL_05_LIP329":"0.0135"},{"Metabolite":"Cer(d18:0/24:2)","ADH1B_KO_01_LIP329":"0.0003","ADH1B_KO_02_LIP329":"0.0003","ADH1B_KO_03_LIP329":"0.0005","ADH1B_KO_04_LIP329":"0.0004","ADH1B_KO_05_LIP329":"0.0005","CTL_01_LIP329":"0.0011","CTL_02_LIP329":"0.0010","CTL_03_LIP329":"0.0011","CTL_04_LIP329":"0.0009","CTL_05_LIP329":"0.0011"},{"Metabolite":"Cer(d18:0/24:3)","ADH1B_KO_01_LIP329":"0.0002","ADH1B_KO_02_LIP329":"0.0003","ADH1B_KO_03_LIP329":"0.0003","ADH1B_KO_04_LIP329":"0.0002","ADH1B_KO_05_LIP329":"0.0004","CTL_01_LIP329":"0.0004","CTL_02_LIP329":"0.0006","CTL_03_LIP329":"0.0005","CTL_04_LIP329":"0.0006","CTL_05_LIP329":"0.0005"},{"Metabolite":"Cer(d18:0/25:0)","ADH1B_KO_01_LIP329":"0.0001","ADH1B_KO_02_LIP329":"0.0001","ADH1B_KO_03_LIP329":"0.0001","ADH1B_KO_04_LIP329":"0.0000","ADH1B_KO_05_LIP329":"0.0001","CTL_01_LIP329":"0.0001","CTL_02_LIP329":"0.0001","CTL_03_LIP329":"0.0001","CTL_04_LIP329":"0.0001","CTL_05_LIP329":"0.0001"},{"Metabolite":"Cer(d18:0/25:1)","ADH1B_KO_01_LIP329":"0.0017","ADH1B_KO_02_LIP329":"0.0022","ADH1B_KO_03_LIP329":"0.0020","ADH1B_KO_04_LIP329":"0.0018","ADH1B_KO_05_LIP329":"0.0022","CTL_01_LIP329":"0.0050","CTL_02_LIP329":"0.0048","CTL_03_LIP329":"0.0054","CTL_04_LIP329":"0.0044","CTL_05_LIP329":"0.0048"},{"Metabolite":"Cer(d18:0/25:2)","ADH1B_KO_01_LIP329":"0.0012","ADH1B_KO_02_LIP329":"0.0020","ADH1B_KO_03_LIP329":"0.0014","ADH1B_KO_04_LIP329":"0.0015","ADH1B_KO_05_LIP329":"0.0017","CTL_01_LIP329":"0.0036","CTL_02_LIP329":"0.0037","CTL_03_LIP329":"0.0047","CTL_04_LIP329":"0.0038","CTL_05_LIP329":"0.0043"},{"Metabolite":"Cer(d18:0/26:0)","ADH1B_KO_01_LIP329":"0.0000","ADH1B_KO_02_LIP329":"0.0001","ADH1B_KO_03_LIP329":"0.0000","ADH1B_KO_04_LIP329":"0.0000","ADH1B_KO_05_LIP329":"0.0001","CTL_01_LIP329":"0.0000","CTL_02_LIP329":"0.0000","CTL_03_LIP329":"0.0001","CTL_04_LIP329":"0.0001","CTL_05_LIP329":"0.0000"},{"Metabolite":"Cer(d18:0/26:1)","ADH1B_KO_01_LIP329":"0.0000","ADH1B_KO_02_LIP329":"0.0003","ADH1B_KO_03_LIP329":"0.0002","ADH1B_KO_04_LIP329":"0.0001","ADH1B_KO_05_LIP329":"0.0001","CTL_01_LIP329":"0.0002","CTL_02_LIP329":"0.0001","CTL_03_LIP329":"0.0002","CTL_04_LIP329":"0.0002","CTL_05_LIP329":"0.0002"},{"Metabolite":"Cer(d18:0/26:2)","ADH1B_KO_01_LIP329":"0.0001","ADH1B_KO_02_LIP329":"","ADH1B_KO_03_LIP329":"","ADH1B_KO_04_LIP329":"0.00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