Summary of Study ST003005
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 PR001873. The data can be accessed directly via it's Project DOI: 10.21228/M8MF0Z 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 | ST003005 |
| Study Title | Brown fat metabolomics of Chow and HFD diet |
| Study Summary | Brown adipose tissue (BAT) metabolites in mice fed a standard diet or a high-fat diet for 12 weeks. N = 6 per group. |
| Institute | Harvard Medical School |
| Last Name | Wang |
| First Name | Dandan |
| Address | 3 Blackfan Circle, Boston, MA, 02115, USA |
| dandanwang2022@gmail.com | |
| Phone | 5083733714 |
| Submit Date | 2023-12-12 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-02-19 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001873 |
| Project DOI: | doi: 10.21228/M8MF0Z |
| Project Title: | Uncoupling Metabolic Health from Thermogenesis via BCAA Flux in Brown Fat |
| Project Type: | MS quantitative analysis |
| Project Summary: | Brown adipose tissue (BAT) is best known for thermogenesis. Whereas numerous studies in rodents found tight associations between the metabolic benefits of BAT and enhanced whole-body energy expenditure, emerging evidence in humans suggests that BAT is protective against Type 2 diabetes independent of body-weight. The underlying mechanism for this dissociation remained unclear. Here, we report that impaired mitochondrial flux of branched-chain amino acids (BCAA) in BAT, by deleting mitochondrial BCAA carrier (MBC, encoded by Slc25a44), was sufficient to cause systemic insulin resistance without affecting whole-body energy expenditure or body-weight. We found that brown adipocytes catabolized BCAAs in the mitochondria as essential nitrogen donors for the biosynthesis of glutamate, N-acetylated amino acids, and one of the products, glutathione. BAT-selective impairment in mitochondrial BCAA flux led to elevated oxidative stress and insulin resistance in the liver, accompanied by reduced levels of BCAA-derived metabolites in the circulation. In turn, supplementation of glutathione restored insulin sensitivity of BAT-specific MBC knockout mice. Notably, a high-fat diet rapidly impaired BCAA catabolism and the synthesis of BCAA-nitrogen derived metabolites in the BAT, while cold-induced BAT activity is coupled with an active synthesis of these metabolites. Together, the present work uncovers a mechanism through which brown fat controls metabolic health independent of thermogenesis via BCAA-derived nitrogen carriers acting on the liver. |
| Institute: | Harvard Medical School |
| Last Name: | Wang |
| First Name: | Dandan |
| Address: | 3 Blackfan Circle, Boston, MA, 02115, USA |
| Email: | dandanwang2022@gmail.com |
| Phone: | 5083733714 |
Subject:
| Subject ID: | SU003119 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 12 weeks |
| Weight Or Weight Range: | 25-30g |
| Gender: | Male |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Diet |
|---|---|---|
| SA326851 | BAT_chow_neg_1 | Chow |
| SA326852 | BAT_chow_neg_6 | Chow |
| SA326853 | BAT_chow_neg_5 | Chow |
| SA326854 | BAT_chow_neg_2 | Chow |
| SA326855 | BAT_chow_neg_3 | Chow |
| SA326856 | BAT_chow_neg_4 | Chow |
| SA326857 | BAT_HFD_neg_6 | HFD |
| SA326858 | BAT_HFD_neg_5 | HFD |
| SA326859 | BAT_HFD_neg_1 | HFD |
| SA326860 | BAT_HFD_neg_2 | HFD |
| SA326861 | BAT_HFD_neg_3 | HFD |
| SA326862 | BAT_HFD_neg_4 | HFD |
| Showing results 1 to 12 of 12 |
Collection:
| Collection ID: | CO003112 |
| Collection Summary: | Animals were sacrificed immediately by cervical dislocation and tissues were rapidly extracted and immediately snap frozen in liquid nitrogen for metabolite profiling. |
| Sample Type: | Brown fat tissue |
Treatment:
| Treatment ID: | TR003128 |
| Treatment Summary: | All mice were housed under a 12 h – 12 h light/dark cycle. Room-temperature mice were housed at 23˚C in ventilated cages with an ACH of 25. Mice were fed a standard diet (Lab Diet 5008) or high-fat diet (Research Diets; D12492; 60% Fat) and had free access to food and water. |
Sample Preparation:
| Sampleprep ID: | SP003125 |
| Sampleprep Summary: | Animals were sacrificed immediately by cervical dislocation and tissues were rapidly extracted and immediately snap frozen in liquid nitrogen for metabolite profiling. For metabolite extraction, tissues were weighed and then homogenized with extraction buffer which consisted of 80% methanol containing D₈-Phenylalanine internal standards at a 40:1 volume to wet weight ratio. Samples were centrifuged at 16,000 x g at 4 °C for 15 min. Finally, 50 μL supernatant was transferred to the glass insert for LC-MS detection. |
Combined analysis:
| Analysis ID | AN004936 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Vanquish |
| Column | Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo orbitrap exploris 240 |
| Ion Mode | NEGATIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH003725 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC BEH Amide (100 x 2.1mm,1.7um) |
| Column Temperature: | 25 °C |
| Flow Gradient: | The linear gradient eluted from 95% B (0.0–1 min), 95% B to 65% B (1–7.0 min), 65% B to 40% B (7.0–8.0 min), 40% B (8.0–9.0 min), 40% B to 95% B (9.0–9.1 min), then stayed at 95% B for 5.9 min. |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 25mM ammonium acetate; 25mM ammonium hydroxide |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004679 |
| Analysis ID: | AN004936 |
| Instrument Name: | Thermo orbitrap exploris 240 |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | ESI source parameters were set as follows: spray voltage, 3500 V or −2800 V, in positive or negative modes, respectively; vaporizer temperature, 350 °C; sheath gas, 50 arb; aux gas, 10 arb; ion transfer tube temperature, 325 °C. The full scan was set as: orbitrap resolution, 60,000; maximum injection time, 100 ms; scan range, 70–1050 Da. The ddMS2 scan was set as: orbitrap resolution, 30,000; maximum injection time, 60 ms; top N setting, 6; isolation width, 1.0 m/z; HCD collision energy (%), 30; Dynamic exclusion mode was set as auto. The data was analyzed by Compound Discoverer 3.3. |
| Ion Mode: | NEGATIVE |
