Summary of Study ST002400
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001546. The data can be accessed directly via it's Project DOI: 10.21228/M8VM64 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002400 |
| 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 |
| jeremie.gautheron@gmail.com | |
| Phone | +33623398373 |
| Submit Date | 2022-12-13 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | wiff |
| Analysis Type Detail | LC-MS |
| Release Date | 2022-12-28 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001546 |
| Project DOI: | doi: 10.21228/M8VM64 |
| 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 |
Subject:
| Subject ID: | SU002489 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Species Group: | Mammals |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype |
|---|---|---|
| SA239098 | ADH1B_KO_03_LIP329 | ADH1B_KO |
| SA239099 | ADH1B_KO_04_LIP329 | ADH1B_KO |
| SA239100 | ADH1B_KO_05_LIP329 | ADH1B_KO |
| SA239101 | ADH1B_KO_02_LIP329 | ADH1B_KO |
| SA239102 | ADH1B_KO_01_LIP329 | ADH1B_KO |
| SA239103 | CTL_02_LIP329 | CTL |
| SA239104 | CTL_03_LIP329 | CTL |
| SA239105 | CTL_04_LIP329 | CTL |
| SA239106 | CTL_05_LIP329 | CTL |
| SA239107 | CTL_01_LIP329 | CTL |
| Showing results 1 to 10 of 10 |
Collection:
| Collection ID: | CO002482 |
| 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 ID: | TR002501 |
| 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. |
Sample Preparation:
| Sampleprep ID: | SP002495 |
| 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. |
Combined analysis:
| Analysis ID | AN003907 | AN003908 | AN003909 | AN003910 | AN003911 |
|---|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS | MS |
| Chromatography type | HILIC | HILIC | HILIC | Reversed phase | Reversed phase |
| Chromatography system | Shimadzu 20AD | Shimadzu 20AD | Shimadzu 20AD | Shimadzu 20AD | Shimadzu 20AD |
| Column | Phenomenex Kinetex HILIC (150 x 3mm,2.6um) | Phenomenex Kinetex HILIC (150 x 3mm,2.6um) | Phenomenex Kinetex HILIC (150 x 3mm,2.6um) | Merck Supelco Ascentis Express C18 (150 x 2.1mm,2.7um) | Merck Supelco Ascentis Express C18 (150 x 2.1mm,2.7um) |
| MS Type | ESI | ESI | ESI | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole | Triple quadrupole | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex 4000 QTrap | ABI Sciex 4000 QTrap | ABI Sciex 4000 QTrap | ABI Sciex 4000 QTrap | ABI Sciex 4000 QTrap |
| Ion Mode | POSITIVE | POSITIVE | POSITIVE | POSITIVE | POSITIVE |
| Units | mol% of total lipids | mol% of total lipids | mol% of total lipids | mol% of total lipids | mol% of total lipids |
Chromatography:
| Chromatography ID: | CH002893 |
| 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; 30 mM ammonium acetate |
| Solvent B: | 100% acetonitrile; 0.2% acetic acid |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002894 |
| Instrument Name: | Shimadzu 20AD |
| Column Name: | Merck Supelco Ascentis Express C18 (150 x 2.1mm,2.7um) |
| Column Temperature: | 43 |
| Flow Gradient: | - |
| Flow Rate: | 300ul/min |
| Solvent A: | 60% acetonitrile/40% water; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 10% acetonitrile/90% isopropanol; 0.1% formic acid; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003646 |
| Analysis ID: | AN003907 |
| Instrument Name: | ABI Sciex 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | MRM acquisition of broad chromatographic peaks of low abundant phospho- and sphingolipid classes: PS, PA, LPC, LPE and some ceramides this acquisition is called "long_1x" |
| Ion Mode: | POSITIVE |
| MS ID: | MS003647 |
| Analysis ID: | AN003908 |
| Instrument Name: | ABI Sciex 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| 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 ID: | MS003648 |
| Analysis ID: | AN003909 |
| Instrument Name: | ABI Sciex 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | MRM acquisition of narrow chromatographic peaks of abundant phospho- and sphingolipid classes: PC and SM following 20-fold dilution This acquisition is referred to as "short_20x" |
| Ion Mode: | POSITIVE |
| MS ID: | MS003649 |
| Analysis ID: | AN003910 |
| Instrument Name: | ABI Sciex 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | MRM acquisition of low abundant neutral lipids: DG This acquisition is referred to as "C18_1x" |
| Ion Mode: | POSITIVE |
| MS ID: | MS003650 |
| Analysis ID: | AN003911 |
| Instrument Name: | ABI Sciex 4000 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | MRM acquisition of abundant neutral lipids: CE and TG after 10 fold dilution This acquisition is referred to as "C18_10x" |
| Ion Mode: | POSITIVE |
