Summary of Study ST003004
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 | ST003004 |
| Study Title | Extracellular fluid metabolomics of BAT and eWAT |
| Study Summary | We quantified metabolites of extracellular fluid samples from BAT and eWAT. Briefly, we collected the BAT_EF samples and eWAT_EF samples from 12 weeks chow diet C57BL/6J mice (n=5). We run the EF metabolomics using high ph HILIC method on Exploris 240. |
| 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: | SU003118 |
| 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 |
| Species Group: | Mammals |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Tissue |
|---|---|---|
| SA326841 | EF_BAT_neg_1 | BAT |
| SA326842 | EF_BAT_neg_5 | BAT |
| SA326843 | EF_BAT_neg_4 | BAT |
| SA326844 | EF_BAT_neg_2 | BAT |
| SA326845 | EF_BAT_neg_3 | BAT |
| SA326846 | EF_eWAT_neg_5 | eWAT |
| SA326847 | EF_eWAT_neg_4 | eWAT |
| SA326848 | EF_eWAT_neg_1 | eWAT |
| SA326849 | EF_eWAT_neg_2 | eWAT |
| SA326850 | EF_eWAT_neg_3 | eWAT |
| Showing results 1 to 10 of 10 |
Collection:
| Collection ID: | CO003111 |
| Collection Summary: | Animals were sacrificed immediately by cervical dislocation and tissues were rapidly extracted. Tissues were subjected to centrifugation (10 min, 800 g, 4°C) following placement in a 20 μm nylon mesh filter (EMD Millipore). |
| Sample Type: | Extracellular fluid |
Treatment:
| Treatment ID: | TR003127 |
| 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) and had free access to food and water. |
Sample Preparation:
| Sampleprep ID: | SP003124 |
| Sampleprep Summary: | Animals were sacrificed immediately by cervical dislocation. Tissues were subjected to centrifugation (10 min, 800 g, 4°C) following placement in a 20 μm nylon mesh filter (EMD Millipore). Metabolites were extracted by adding extraction buffer at a ratio of 1:100 interstitial fluid to methanol. Samples were then centrifuged twice (5 min, 10,000 g, 4°C) and supernatant was collected. |
Combined analysis:
| Analysis ID | AN004935 |
|---|---|
| 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: | CH003724 |
| 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: | MS004678 |
| Analysis ID: | AN004935 |
| 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 |
