Summary of Study ST003175
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 PR001974. The data can be accessed directly via it's Project DOI: 10.21228/M8K13N This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST003175 |
| Study Title | Inactivation of mitochondrial MUL1 E3 ubiquitin ligase inhibits lipogenesis and prevents diet-induced obesity in mice |
| Study Type | Basic Research |
| Study Summary | Characterize the role of mitochondrial MUL1 E3-ubiquitin ligase on energy metabolism and lipogenesis using Mul1 deficient, Mul1(-/-), mice. MUL1 protein is involved in mitochondrial dynamics, and innate immune response but its primary function might be the regulation of lipogenesis under conditions of nutritional overload. Using metabolic cages, we monitored whole body energy expenditure, metabolism, and thermoregulation of the Mul1(-/-) mice under standard diet (ND) or high fat diet (HFD). We examined the effect of Mul1 inactivation on body weight, HFD-induced adiposity, fatty liver, glucose intolerance, and insulin resistance. We performed global metabolomics, lipidomic, and genome-wide mRNA sequencing using liver from Mul1(+/+) and Mul1(-/-) animals on HFD. The expression level of key proteins involved in lipogenesis and their regulation in the absence of MUL1 was monitored by SDS-PAGE and Western blot analysis. We found that Mul1(-/-) animals have a metabolic phenotype that confers robust resistance to HFD-induced obesity. Several metabolic and lipidomic pathways are perturbed in the liver of Mul1(-/-) animals on HFD, particularly the one driven by Stearoyl-CoA Desaturase 1 (SCD1), a key regulator of lipid metabolism and obesity. In addition, key enzymes involved in lipogenesis and fatty acid oxidation such as ACC1, FASN, AMPK, and CTP1 were also modulated. The concerted deregulation of these enzymes, in the absence of MUL1, causes reduced fat storage and increased fatty acid oxidation. We identified a new function of mitochondrial MUL1 E3 ubiquitin ligase in the regulation of lipogenesis and adiposity, particularly during conditions of HFD. Inactivation of MUL1 provides resistance to HFD-induced obesity and can be a therapeutic target for the treatment of obesity, type 2 diabetes, and nonalcoholic fatty liver disease (NAFLD). |
| Institute | University of Florida |
| Last Name | Merritt |
| First Name | Matthew |
| Address | SW ARCHER RD, Gainesville, FL 32611 |
| rmahar@hnbgu.ac.in | |
| Phone | 9557559849 |
| Submit Date | 2024-04-10 |
| Num Groups | 2 |
| Total Subjects | 12 |
| Num Males | 6 |
| Num Females | 6 |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-04-19 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001974 |
| Project DOI: | doi: 10.21228/M8K13N |
| Project Title: | Inactivation of mitochondrial MUL1 E3 ubiquitin ligase inhibits lipogenesis and prevents diet-induced obesity in mice |
| Project Type: | Basic Research |
| Project Summary: | Characterize the role of mitochondrial MUL1 E3-ubiquitin ligase on energy metabolism and lipogenesis using Mul1 deficient, Mul1(-/-), mice. MUL1 protein is involved in mitochondrial dynamics, and innate immune response but its primary function might be the regulation of lipogenesis under conditions of nutritional overload. Using metabolic cages, we monitored whole body energy expenditure, metabolism, and thermoregulation of the Mul1(-/-) mice under standard diet (ND) or high fat diet (HFD). We examined the effect of Mul1 inactivation on body weight, HFD-induced adiposity, fatty liver, glucose intolerance, and insulin resistance. We performed global metabolomics, lipidomic, and genome-wide mRNA sequencing using liver from Mul1(+/+) and Mul1(-/-) animals on HFD. The expression level of key proteins involved in lipogenesis and their regulation in the absence of MUL1 was monitored by SDS-PAGE and Western blot analysis. We found that Mul1(-/-) animals have a metabolic phenotype that confers robust resistance to HFD-induced obesity. Several metabolic and lipidomic pathways are perturbed in the liver of Mul1(-/-) animals on HFD, particularly the one driven by Stearoyl-CoA Desaturase 1 (SCD1), a key regulator of lipid metabolism and obesity. In addition, key enzymes involved in lipogenesis and fatty acid oxidation such as ACC1, FASN, AMPK, and CTP1 were also modulated. The concerted deregulation of these enzymes, in the absence of MUL1, causes reduced fat storage and increased fatty acid oxidation. We identified a new function of mitochondrial MUL1 E3 ubiquitin ligase in the regulation of lipogenesis and adiposity, particularly during conditions of HFD. Inactivation of MUL1 provides resistance to HFD-induced obesity and can be a therapeutic target for the treatment of obesity, type 2 diabetes, and nonalcoholic fatty liver disease (NAFLD). |
| Institute: | University of Florida |
| Department: | Biochemistry |
| Laboratory: | Merritt's Lab |
| Last Name: | Merritt |
| First Name: | Matthew |
| Address: | SW ARCHER RD, Gainesville, FL 32611 |
| Email: | rmahar@hnbgu.ac.in |
| Phone: | 9557559849 |
| Funding Source: | NIH |
| Contributors: | Lucia Cilenti, Jacopo Di Gregorio, Rohit Mahar, Fei Liu, Camilla T. Ambivero, Muthu Periasamy, Matthew E. Merrit, and Antonis S. Zervos |
Subject:
| Subject ID: | SU003294 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | Wild type and Mul1(+/+) mice |
| Age Or Age Range: | 8 weeks |
| Gender: | Male and female |
| Animal Light Cycle: | 12-hour/12-hour light/dark cycle |
| Animal Feed: | Research Diets D12492 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype | Sample source |
|---|---|---|---|
| SA343203 | Mul1(+/+)_TZ 34 | Mul1(+/+) | Liver |
| SA343204 | Mul1(+/+)_TZ 37 | Mul1(+/+) | Liver |
| SA343205 | Mul1(+/+)_TZ 36 | Mul1(+/+) | Liver |
| SA343206 | Mul1(+/+)_TZ 35 | Mul1(+/+) | Liver |
| SA343207 | Mul1(-/-)_TZ 41 | Mul1(-/-) | Liver |
| SA343208 | Mul1(-/-)_TZ 38 | Mul1(-/-) | Liver |
| SA343209 | Mul1(-/-)_TZ 39 | Mul1(-/-) | Liver |
| SA343210 | Mul1(-/-)_TZ 40 | Mul1(-/-) | Liver |
| Showing results 1 to 8 of 8 |
Collection:
| Collection ID: | CO003287 |
| Collection Summary: | Liver tissue was obtained from Mul1(+/+) and Mul1(-/-) mice following a HFD for 16 weeks and stained with H&E or Oil Red Oil. |
| Sample Type: | Liver |
| Storage Conditions: | -20℃ |
Treatment:
| Treatment ID: | TR003303 |
| Treatment Summary: | Liver tissue was obtained from Mul1(+/+) and Mul1(-/-) mice following a HFD for 16 weeks and stained with H&E or Oil Red Oil. |
Sample Preparation:
| Sampleprep ID: | SP003301 |
| Sampleprep Summary: | liver tissues from mice were fixed overnight in 10% neutral formalin and embedded in paraffin. Paraffin-embedded tissues were cut into sections and stained with hematoxylin and eosin (H&E) for assessment of liver histology. Liver tissue from Mul1(+/+) and Mul1(-/-) mice on HFD were subjected to the Folch extraction in order to separate lipids and polar metabolites. |
| Processing Storage Conditions: | 4℃ |
| Extract Storage: | -20℃ |
Combined analysis:
| Analysis ID | AN005211 | AN005212 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | A.U. | A.U. |
Chromatography:
| Chromatography ID: | CH003942 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0−0.5 min, 60% B; 0.5−2 min, 60−95% B; 2−17 min, 95% B; 17−17.5 min, 95−60% B; 17.5−20 min, 60% B |
| Flow Rate: | 350 µl/min |
| Solvent A: | 60:40 acetonitrile:water; 10mM ammonium formate; 0.1% formic acid |
| Solvent B: | 90:8:2 2-propanol:acetonitrile:water; 10mM ammonium formate; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004944 |
| Analysis ID: | AN005211 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The aqueous phase was analyzed using the Thermo Q-Exactive Orbitrap mass spectrometer with Dionex UHPLC and autosampler. All samples were analyzed in positive and negative simultaneously with heated electrospray ionization (HESI) with a mass resolution of 35,000 at m/z 200 as separate injections. The polar metabolites were analyzed on Thermo Q-Exactive Orbitrap mass spectrometer equipped with UHPLC. Data were analyzed with MZmine and features were aligned for identification across samples. The metabolites were searched against the Southeastern Center for Integrated Metabolomics (SECIM) metabolite library using retention time and corresponding mass spectral data. Lipids isolated from Folch extraction procedure were also analyzed by the same instrument. Lipidomics data were analyzed using the LipidMatch software and identified lipid entities were exported in a tabular form. |
| Ion Mode: | POSITIVE |
| MS ID: | MS004945 |
| Analysis ID: | AN005212 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The aqueous phase was analyzed using the Thermo Q-Exactive Orbitrap mass spectrometer with Dionex UHPLC and autosampler. All samples were analyzed in positive and negative simultaneously with heated electrospray ionization (HESI) with a mass resolution of 35,000 at m/z 200 as separate injections. The polar metabolites were analyzed on Thermo Q-Exactive Orbitrap mass spectrometer equipped with UHPLC. Data were analyzed with MZmine and features were aligned for identification across samples. The metabolites were searched against the Southeastern Center for Integrated Metabolomics (SECIM) metabolite library using retention time and corresponding mass spectral data. Lipids isolated from Folch extraction procedure were also analyzed by the same instrument. Lipidomics data were analyzed using the LipidMatch software and identified lipid entities were exported in a tabular form. |
| Ion Mode: | NEGATIVE |
