Summary of Study ST003785
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 PR002362. The data can be accessed directly via it's Project DOI: 10.21228/M89Z6M 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 | ST003785 |
| Study Title | Sex differences in lipid profiles of visceral adipose tissue with obesity and gonadectomy |
| Study Type | Lipidomics |
| Study Summary | In obesity, adipose tissue (AT) expansion is accompanied by chronic inflammation. Altered lipid composition in the visceral or gonadal white AT (GWAT) directly drive AT macrophage (ATM) accumulation and activation, switching immune cells to a proinflammatory phenotype. Sex steroid hormones modulate sex differences in visceral vs. subcutaneous lipid accumulation that correlates with metabolic syndrome, especially in men and post-menopausal women who are more prone to truncal obesity. Prior studies demonstrated sex differences in lipid species in HFD-fed mice but there is a gap in understanding the role of sex hormones in these lipid species. We hypothesized that sex hormone alterations with gonadectomy (GX) would further impact lipid composition in the obese GWAT. We performed untargeted lipidomics on GWAT from castrated (CAS) male (M) mice and ovariectomized (OVX) female (F) mice. The control groups are named M SHAM HFD and F SHAM HFD, and the experimental groups are named M CAS HFD (or M GX HFD in text) and F OVX HFD (or F GX HFD in text). Untargeted lipidomics of obese GWAT identified sex differences in phospholipids, sphingolipids, sterols, fatty acyls, saccharo-lipids and prenol- lipids. Males had significantly higher content of key precursor fatty acids (palmitic, oleic, linoleic and arachidonic acid) when compared to female and GX mice. Bulk RNA sequencing of sorted GWAT ATMs highlighted sex and diet differences in PUFA metabolism genes and oxylipin genes. These findings of sexual dimorphism in stored lipid species emphasize sex-differences in GWAT lipid metabolism pathways that alter lipid composition driving inflammation responses and metabolic disease risk. |
| Institute | University of Michigan |
| Department | BRCF |
| Laboratory | Metabolomics core |
| Last Name | Kachman |
| First Name | Maureen |
| Address | 1000 Wall St. |
| mkachman@med.umich.edu | |
| Phone | 734-232-0842 |
| Submit Date | 2025-02-27 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | wiff |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-03-31 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002362 |
| Project DOI: | doi: 10.21228/M89Z6M |
| Project Title: | Study the role of high fat diet and sex hormone fluctuations on visceral or gonadal adipose tissue (GWAT) lipid species |
| Project Type: | MS |
| Project Summary: | This study investigated effects of high fat diet and sex hormone alterations with gonadectomy (GX) on lipid composition in the obese GWAT. Untargeted lipidomics was performed on GWAT from castrated (CAS) male (M) mice and ovariectomized (OVX) female (F) mice. The control groups are named M SHAM HFD and F SHAM HFD, and the experimental groups are named M CAS HFD (or M GX HFD in text) and F OVX HFD (or F GX HFD in text). Untargeted lipidomics of obese GWAT identified sex differences in phospholipids, sphingolipids, sterols, fatty acyls, saccharo-lipids and prenol- lipids. Males had significantly higher content of key precursor fatty acids (palmitic, oleic, linoleic and arachidonic acid) when compared to female and GX mice. These findings of sexual dimorphism in stored lipid species emphasize sex-differences in GWAT lipid metabolism pathways that alter lipid composition driving inflammation responses and metabolic disease risk. |
| Institute: | University of Michigan |
| Department: | BRCF |
| Laboratory: | Metabolomics core |
| Last Name: | Kachman |
| First Name: | Maureen |
| Address: | 1000 Wall St. |
| Email: | mkachman@med.umich.edu |
| Phone: | 734-232-0842 |
Subject:
| Subject ID: | SU003919 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Group | Sex |
|---|---|---|---|
| SA411096 | Singer_GWAT_G7 | Castration (CAS) | male |
| SA411097 | Singer_GWAT_G8 | Castration (CAS) | male |
| SA411098 | Singer_GWAT_G9 | Castration (CAS) | male |
| SA411099 | Singer_GWAT_G10 | Castration (CAS) | male |
| SA411100 | Singer_GWAT_G11 | Castration (CAS) | male |
| SA411101 | Singer_GWAT_G12 | Castration (CAS) | male |
| SA411102 | Singer_GWAT_G14 | F SHAM HFD | female |
| SA411103 | Singer_GWAT_G18 | F SHAM HFD | female |
| SA411104 | Singer_GWAT_G17 | F SHAM HFD | female |
| SA411105 | Singer_GWAT_G16 | F SHAM HFD | female |
| SA411106 | Singer_GWAT_G15 | F SHAM HFD | female |
| SA411107 | Singer_GWAT_G13 | F SHAM HFD | female |
| SA411108 | Singer_GWAT_G2 | M_SHAM HFD | male |
| SA411109 | Singer_GWAT_G6 | M_SHAM HFD | male |
| SA411110 | Singer_GWAT_G5 | M_SHAM HFD | male |
| SA411111 | Singer_GWAT_G4 | M_SHAM HFD | male |
| SA411112 | Singer_GWAT_G3 | M_SHAM HFD | male |
| SA411113 | Singer_GWAT_G1 | M_SHAM HFD | male |
| SA411114 | Singer_GWAT_G19 | Ovariectomy (OVX) | female |
| SA411115 | Singer_GWAT_G20 | Ovariectomy (OVX) | female |
| SA411116 | Singer_GWAT_G21 | Ovariectomy (OVX) | female |
| SA411117 | Singer_GWAT_G22 | Ovariectomy (OVX) | female |
| SA411118 | Singer_GWAT_G23 | Ovariectomy (OVX) | female |
| SA411119 | Singer_GWAT_G24 | Ovariectomy (OVX) | female |
| Showing results 1 to 24 of 24 |
Collection:
| Collection ID: | CO003912 |
| Collection Summary: | Gonadal white adipose tissue (GWAT) was excised from mice at the end of 16 weeks of high fat diet (HFD) |
| Sample Type: | Adipose tissue |
Treatment:
| Treatment ID: | TR003928 |
| Treatment Summary: | This study investigated effects of high fat diet and sex hormone alterations with gonadectomy (GX) on lipid composition in the obese GWAT. C57Bl/6J (000664) gonadectomized (GX) and sham surgery animals were purchased from Jackson Laboratories at 4 weeks of age. Mice were gonadectomized at 3-4 weeks of age. All mice were fed ad libitum 13.5% fat (5LOD; LabDiet) or HFD consisting of 60% of calories from fat (Research Diets; D12492), starting at 6 weeks of age for 16 weeks of duration. GWAT was excised at the end of 16 weeks of HFD. Untargeted lipidomics was then performed on GWAT from castrated (CAS) male (M) mice, ovariectomized (OVX) female (F) mice and control sham surgery - M SHAM and F SHAM (Total 4 groups; 6 per group). The control groups are named M SHAM HFD and F SHAM HFD, and the experimental groups are named M CAS HFD (or M GX HFD in text) and F OVX HFD (or F GX HFD in text). |
Sample Preparation:
| Sampleprep ID: | SP003925 |
| Sampleprep Summary: | Standard shotgun lipidomics sample preparation protocol, no derivatization |
| Sampleprep Protocol Filename: | A004_Shotgun_Lipidomics.PDF |
Combined analysis:
| Analysis ID | AN006219 | AN006220 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Shimadzu Nexera X2 | Shimadzu Nexera X2 |
| Column | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI | ESI |
| MS instrument type | Triple TOF | Triple TOF |
| MS instrument name | ABI Sciex 5600+ TripleTOF | ABI Sciex 5600+ TripleTOF |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | Normalised counts | Normalised counts |
Chromatography:
| Chromatography ID: | CH004715 |
| Chromatography Summary: | Negative mode |
| Methods Filename: | EX01123-LC-method.pdf |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 55 |
| Flow Gradient: | (Time : Pump B%) (0.01 : 40) (10.00 : 98) (17.00 : 98) (17.10 : 40) |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | 40% Acetonitrile/60% water; 10 mM ammonium acetate |
| Solvent B: | 10% Acetonitrile/5% water/85% isopropanol; 10 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH004716 |
| Chromatography Summary: | Positive mode |
| Methods Filename: | EX01123-LC-method.pdf |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 55 |
| Flow Gradient: | (Time : Pump B%) (0.01 : 40) (10.00 : 98) (17.00 : 98) (17.10 : 40) |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | Acetonitrile 40%/60% water; 10 mM ammonium acetate |
| Solvent B: | 10% Acetonitrile/5% water/85% isopropanol; 10 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS005923 |
| Analysis ID: | AN006219 |
| Instrument Name: | ABI Sciex 5600+ TripleTOF |
| Instrument Type: | Triple TOF |
| MS Type: | ESI |
| MS Comments: | Data Dependent LC-MS/MS Analysis Chromatographic separation was performed on a Shimadzu CTO-20A Nexera X2 UHPLC systems equipped with a degasser, binary pump, thermostatted autosampler, and column oven (all from Shimadzu). The column heater temperature was maintained at 55oC. The injection volume was 5 μL for all analyses. For lipid separation, the lipid extract is injected onto a 1.8 μm particle 50 × 2.1 mm id Waters Acquity HSS T3 column (Waters, Milford, MA) which is heated to 55°C. Elution is performed using acetonitrile / water (40:60, v/v) with 10 mM ammonium acetate as solvent A and acetonitrile / water / isopropanol (10 : 5 : 85 v/v) with 10 mM ammonium acetate as solvent B. Column is equilibrated for 3 min before the next injection making total run time 14 min. The flow rate was 0.400uL/min. The data acquisition of each sample was performed in both positive and negative ionization modes, using a TripleTOF 5600 equipped with a Turbo VTM ion source (AB Sciex, Concord, Canada). Column effluent was directed to the ESI source. The source voltage was set to 5500V for positive ionization and 4500V for negative ionization mode. The declustering potential (DP) was 60 V and source temperature was 450oC for both modes. The curtain gas flow, nebulizer, and heater gas were set to 30, 40, and 45 arbitrary units. The instrument was set to perform one TOF MS survey scan (150 ms) and 15 MS/MS scans with a total duty cycle time of 2.4 s. The mass range of both mode was 50-1200 m/z. Acquisition of MS/MS spectra was controlled by data dependent acquisition (DDA) function of the Analyst TF software (AB Sciex, Concord, Canada) with application of following parameters—dynamic background subtraction, charge monitoring to exclude multiply charged ions and isotopes, and dynamic exclusion of former target ions for 9 s. Rolling collision energy was set whereby the software calculated the CE value to be applied as a function of m/z. Mass accuracy was maintained by the use of an automated calibrant delivery system (AB Sciex, Concord, Canada) interfaced to the second inlet of the DuoSpray source. Calibrations were performed at the start of a workday or whenever ionization polarity was changed. Pooled human plasma sample and pooled experimental sample (prepared by combining small aliquots of all client’s samples) are used to control the quality of sample preparation and analysis. Randomization scheme is used to distribute pooled samples within the set. Mixture of pure authentic standards is used to monitor the instrument performance on a regular basis. Data analysis Lipids are identified using LIPIDBLAST package (http://fiehnlab.ucdavis.edu/projects/LipidBlast) - computer-generated tandem MS library of 212,516 spectra covering 119,200 compounds representing 26 lipid classes, including phospholipids, glycerolipids, bacterial lipoglycans and plant glycolipids. Quantification of lipids is done by Multiquant software (AB-SCIEX). Normalization of the data for different lipid classes is performed based on a set of internal standards. |
| Ion Mode: | NEGATIVE |
| Acquisition Parameters File: | EX01123-NegativeMode-Acquisition Info.pdf |
| Analysis Protocol File: | A004_Shotgun_Lipidomics.PDF |
| MS ID: | MS005924 |
| Analysis ID: | AN006220 |
| Instrument Name: | ABI Sciex 5600+ TripleTOF |
| Instrument Type: | Triple TOF |
| MS Type: | ESI |
| MS Comments: | Data Dependent LC-MS/MS Analysis Chromatographic separation was performed on a Shimadzu CTO-20A Nexera X2 UHPLC systems equipped with a degasser, binary pump, thermostatted autosampler, and column oven (all from Shimadzu). The column heater temperature was maintained at 55oC. The injection volume was 5 μL for all analyses. For lipid separation, the lipid extract is injected onto a 1.8 μm particle 50 × 2.1 mm id Waters Acquity HSS T3 column (Waters, Milford, MA) which is heated to 55°C. Elution is performed using acetonitrile / water (40:60, v/v) with 10 mM ammonium acetate as solvent A and acetonitrile / water / isopropanol (10 : 5 : 85 v/v) with 10 mM ammonium acetate as solvent B. Column is equilibrated for 3 min before the next injection making total run time 14 min. The flow rate was 0.400uL/min. The data acquisition of each sample was performed in both positive and negative ionization modes, using a TripleTOF 5600 equipped with a Turbo VTM ion source (AB Sciex, Concord, Canada). Column effluent was directed to the ESI source. The source voltage was set to 5500V for positive ionization and 4500V for negative ionization mode. The declustering potential (DP) was 60 V and source temperature was 450oC for both modes. The curtain gas flow, nebulizer, and heater gas were set to 30, 40, and 45 arbitrary units. The instrument was set to perform one TOF MS survey scan (150 ms) and 15 MS/MS scans with a total duty cycle time of 2.4 s. The mass range of both mode was 50-1200 m/z. Acquisition of MS/MS spectra was controlled by data dependent acquisition (DDA) function of the Analyst TF software (AB Sciex, Concord, Canada) with application of following parameters—dynamic background subtraction, charge monitoring to exclude multiply charged ions and isotopes, and dynamic exclusion of former target ions for 9 s. Rolling collision energy was set whereby the software calculated the CE value to be applied as a function of m/z. Mass accuracy was maintained by the use of an automated calibrant delivery system (AB Sciex, Concord, Canada) interfaced to the second inlet of the DuoSpray source. Calibrations were performed at the start of a workday or whenever ionization polarity was changed. Pooled human plasma sample and pooled experimental sample (prepared by combining small aliquots of all client’s samples) are used to control the quality of sample preparation and analysis. Randomization scheme is used to distribute pooled samples within the set. Mixture of pure authentic standards is used to monitor the instrument performance on a regular basis. Data analysis Lipids are identified using LIPIDBLAST package (http://fiehnlab.ucdavis.edu/projects/LipidBlast) - computer-generated tandem MS library of 212,516 spectra covering 119,200 compounds representing 26 lipid classes, including phospholipids, glycerolipids, bacterial lipoglycans and plant glycolipids. Quantification of lipids is done by Multiquant software (AB-SCIEX). Normalization of the data for different lipid classes is performed based on a set of internal standards. |
| Ion Mode: | POSITIVE |
| Acquisition Parameters File: | EX01123-PositiveMode-Acquisition Info.pdf |
| Analysis Protocol File: | A004_Shotgun_Lipidomics.PDF |
