Summary of Study ST004281
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 PR002705. The data can be accessed directly via it's Project DOI: 10.21228/M80V75 This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST004281 |
| Study Title | 2DLC-MS-based lipidomics of alcohol-associated liver disease mouse liver and brain |
| Study Type | Original research |
| Study Summary | To investigate lipidome dysfunctions, the lipidomes of liver and brain tissues from alcohol-fed (AF) and pair-fed (PF) mice were characterized by comprehensive two-dimensional liquid chromatography-mass spectrometry. |
| Institute | University of Louisville |
| Department | Chemistry |
| Laboratory | Dr. Xiang Zhang lab |
| Last Name | Feng |
| First Name | Jing |
| Address | 2210 S Brook St. Louisville, KY 40208 |
| jing.feng@louisville.edu | |
| Phone | 5026187846 |
| Submit Date | 2025-10-06 |
| Num Groups | 2 |
| Total Subjects | 15 |
| Num Females | 15 |
| Publications | Journal of Proteome Research, Submitted |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2026-01-02 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002705 |
| Project DOI: | doi: 10.21228/M80V75 |
| Project Title: | Impact of alcohol exposure on the liver and brain lipidomes of mice with experimental alcohol-associated liver disease |
| Project Type: | Original research |
| Project Summary: | Alcohol-associated liver disease (ALD) has imposed a substantial public health burden in the United States, and alcohol consumption induces systemic lipid dysregulation in ALD. To investigate this, the lipidomes of liver and brain tissues from alcohol-fed (AF) and pair-fed (PF) mice were characterized by comprehensive two-dimensional liquid chromatography-mass spectrometry. Multiple database matching was used for high-confidence lipid identification. Univariable and multivariable analyses were employed to identify lipids with significantly altered abundance between AF and PF mice. Furthermore, differential correlation analysis and cross-tissue co-dysregulation mapping were performed. Our data indicated that alcohol feeding induced profound tissue-specific lipidomic alterations. Triglycerides increased significantly, and phospholipids decreased in the livers of alcohol-treated mice, whereas their brains exhibited elevation of oxidized lipids with subclass-divergent changes in glycerolipids, glycerophospholipids, and sphingolipids. Differential correlation analysis revealed extensive remodeling of lipid interaction networks in the alcohol-exposed brain. Cross-tissue analysis identified 18 co-dysregulated lipids demonstrating a strongly correlated lipid regulation shift in AF mice. Collectively, alcohol rewires organ-specific lipidomes through distinct mechanisms and induces correlated multi-tissue lipid dysregulation, suggesting potential lipid-centric communication along the liver-brain axis. These findings provide novel insights into ALD pathogenesis and reveal potential biomarkers for multi-organ involvement. |
| Institute: | University of Louisville |
| Department: | Chemistry |
| Laboratory: | Dr. Xiang Zhang lab |
| Last Name: | Feng |
| First Name: | Jing |
| Address: | 2210 S brook St. Louisville, KY 40208 |
| Email: | jing.feng@louisville.edu |
| Phone: | 5026187846 |
| Funding Source: | This work was supported by NIH [1P20GM113226 (CJM); 1P50AA024337 (CJM); 1U01AA026934 (CJM); 1U01AA026926 (CJM); 1U01AA026980 (CJM); 1R21AA031563 (LH); and S10OD020106 (XZ)]. This work was also supported by the Department of Veterans Affairs, 1I01BX002996-01A2 (CJM). |
| Publications: | Journal of Proteome Research, Submitted |
| Contributors: | Yuan Fang |
Subject:
| Subject ID: | SU004434 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | wild-type |
| Age Or Age Range: | 12 weeks |
| Gender: | Female |
| Animal Housing: | Biorepository of the University of Louisville Hepatology and Toxicology COBRE |
| Animal Light Cycle: | 12h |
| Animal Feed: | high-fat (60%) diets, diets were purchased from the Research Diets (D12492i, New Brunswick, NJ, USA). |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Treatment |
|---|---|---|---|
| SA498326 | brain_329 | brain | Alcohol |
| SA498327 | brain_321 | brain | Alcohol |
| SA498328 | brain_330 | brain | Alcohol |
| SA498329 | brain_320 | brain | Alcohol |
| SA498330 | brain_328 | brain | Alcohol |
| SA498331 | brain_322 | brain | Alcohol |
| SA498332 | brain_323 | brain | Alcohol |
| SA498333 | brain_327 | brain | Alcohol |
| SA498334 | brain_348 | brain | Control |
| SA498335 | brain_347 | brain | Control |
| SA498336 | brain_346 | brain | Control |
| SA498337 | brain_345 | brain | Control |
| SA498338 | brain_316 | brain | Control |
| SA498339 | brain_315 | brain | Control |
| SA498340 | brain_319 | brain | Control |
| SA498341 | liver_329 | liver | Alcohol |
| SA498342 | liver_328 | liver | Alcohol |
| SA498343 | liver_327 | liver | Alcohol |
| SA498344 | liver_323 | liver | Alcohol |
| SA498345 | liver_322 | liver | Alcohol |
| SA498346 | liver_321 | liver | Alcohol |
| SA498347 | liver_320 | liver | Alcohol |
| SA498348 | liver_330 | liver | Alcohol |
| SA498349 | liver_315 | liver | Control |
| SA498350 | liver_316 | liver | Control |
| SA498351 | liver_319 | liver | Control |
| SA498352 | liver_345 | liver | Control |
| SA498353 | liver_346 | liver | Control |
| SA498354 | liver_347 | liver | Control |
| SA498355 | liver_348 | liver | Control |
| Showing results 1 to 30 of 30 |
Collection:
| Collection ID: | CO004427 |
| Collection Summary: | Samples for lipidomic analysis were obtained from the biorepository of the University of Louisville Hepatology and Toxicology COBRE. The study design involved wild type C57BL6/J female mice (8-week-old, bred in-house), which were fed high-fat (60%) diets for 12 weeks. Diets were purchased from the Research Diets (D12492i, New Brunswick, NJ, USA). After 12 weeks high fat diet feeding, mice were gavaged with vehicle (saline) or 31.5% (v/v) ethanol (in saline) at 5 g/kg. Eight mice received a single dose of ethanol (5 g/kg) by oral gavage, and seven mice received the same amount of saline as control. The mice with ethanol treatment were grouped into alcohol-fed (AF, n = 8) and control, as pair-fed (PF, n = 7). Nine hours later, mice were deeply anesthetized with 100 mg/kg ketamine and 16 mg/kg xylazine. Following collection of blood from the inferior vena cava, organs were collected, snap-frozen in liquid nitrogen, and stored at -80⁰C. |
| Collection Protocol ID: | IACUC 15423 to Irina Kirpich |
| Sample Type: | Liver; Brain |
| Collection Location: | Clinical Translational Research Building, University of Louisville Health Sciences Campus, Louisville, Kentucky. |
| Collection Frequency: | Once (endpoint) |
| Collection Duration: | 5 min |
| Volumeoramount Collected: | whole organs |
| Storage Conditions: | -80℃ |
| Collection Vials: | 1.5 mL self-standing microcentrifuge tubes with O-ring and screw top |
| Storage Vials: | 1.5 mL self-standing microcentrifuge tubes with O-ring and screw top |
| Collection Tube Temp: | room temperature |
| Additives: | 10 U/mL sodium heparin to blood samples |
Treatment:
| Treatment ID: | TR004443 |
| Treatment Summary: | On the final day of the dietary intervention, eight mice received a single dose of ethanol (5 g/kg) by oral gavage, and seven mice received the same amount of saline as control. In this study only control mice were used. Fat-1 transgenic mice were assayed in study ST004300. |
| Treatment Protocol ID: | IACUC 15423 |
| Treatment: | Ethanol binge |
| Treatment Compound: | Ethanol |
| Treatment Route: | Oral |
| Treatment Dose: | 5 g ethanol/kg body weight |
| Treatment Dosevolume: | 20 µl of 31.5% (v/v) ethanol solution/g body weight |
| Treatment Vehicle: | Pharmaceutical (USP)-grade saline |
| Animal Anesthesia: | Endpoint only |
| Animal Acclimation Duration: | 12 weeks (bred in-house) |
| Animal Endp Euthanasia: | 100 mg/kg ketamine+16mg/kg xylazine |
| Animal Endp Tissue Coll List: | Liver, brain |
| Animal Endp Clinical Signs: | >10% body weight loss, body condition score, facial grimace scale for pain and distress. |
Sample Preparation:
| Sampleprep ID: | SP004440 |
| Sampleprep Summary: | Lipids were extracted from mouse liver and brain tissues using a modified Folch method [Folch, et. al., J. Biol. Chem. 1957;226:497–509. doi: 10.1016/S0021-9258(18)64849-5]. Briefly, tissues were first homogenized in methanol (mg: μL, 3:20). Then, 200 μL homogenate was mixed with 400 μL of chloroform, vortexed for 2 min, and incubated for 1 h. Subsequently, 120 μL of deionized water was added, and the mixture was vortexed again for 2 min, followed by centrifugation at 5000 rpm for 5 min at room temperature. The chloroform phase (300 μL) in the bottom layer was collected and dried using nitrogen flow. The dried sample was then redissolved in 150 μL of chloroform/methanol (v/v, 2:1) with 1 mM butylated hydroxytoluene (BHT) and transferred to a glass vial for 2DLC-MS analysis. |
| Extraction Method: | Folch method |
| Extract Enrichment: | Chloroform |
| Extract Storage: | -80℃ |
| Sample Derivatization: | N/A |
| Sample Spiking: | N/A |
Combined analysis:
| Analysis ID | AN007120 | AN007121 |
|---|---|---|
| Chromatography ID | CH005410 | CH005411 |
| MS ID | MS006816 | MS006817 |
| Analysis type | MS | MS |
| Chromatography type | HILIC | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
| Column | Waters ACQUITY UPLC BEH HILIC (150 x 2.1 mm, 1.7 μm) | Waters ACQUITY UPLC BEH C18 (50 x 2.1 mm, 1.7 μm) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | UNSPECIFIED | UNSPECIFIED |
| Units | Relative Abundance | Relative Abundance |
Chromatography:
| Chromatography ID: | CH005410 |
| Chromatography Summary: | Lipid profiling was performed on a Dionex UltiMate™ 3000 UHPLC system.Lipids eluted from the HILIC column were collected into five fractions based on their retention time (Rt) using a 10-port divert valve and two Acquity BEH C18 trap columns (Waters Corporation). |
| Methods Filename: | Chromatography_method.pdf |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters ACQUITY UPLC BEH HILIC (150 x 2.1 mm, 1.7 μm) |
| Column Temperature: | 55⁰C |
| Flow Gradient: | 0-5 min, 0%; 5-90 min, 0% to 90%; 90-95 min, 90% to 0% |
| Flow Rate: | 0.15 mL/min |
| Solvent A: | 97.5% acetonitrile/2.5% water; 10 mM ammonium formate |
| Solvent B: | 50% methanol/50% water; 10 mM ammonium formate |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH005411 |
| Chromatography Summary: | Each HILIC fraction was then subjected to an RPC column, Acquity UPLC BEH C18 column (50 × 2.1 mm, 1.8 μm, Waters Corporation) for further separation |
| Methods Filename: | Chromatography_method.pdf |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters ACQUITY UPLC BEH C18 (50 x 2.1 mm, 1.7 μm) |
| Column Temperature: | 55⁰C |
| Flow Gradient: | 0-5 min, 0%; 5-53 min, 8%; 53-73 min, 8% to 30%; 73-77 min 30% to 60%; 77.1-95 min, 60% to 0% |
| Flow Rate: | 0.5 mL/min |
| Solvent A: | 60% acetonitrile/40% water; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/7.5% acetonitrile/2.5% water; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS006816 |
| Analysis ID: | AN007120 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The mass spectrometer was operated in both positive and negative modes with a scan range 200-1500 m/z under 30,000 resolution at 200 m/z. Electrospray ionization (ESI) settings were: sheath gas 50, aux gas 13, spray voltage 3.5 kV, capillary temperature 263°C, S-lens RF 55, and heater temperature 425°C. MS/MS data acquisition settings were: microscans 2, resolution 15,000 at 200 m/z, maximum injection time 100 ms, and collision energies 20 and 30 eV. |
| Ion Mode: | UNSPECIFIED |
| Analysis Protocol File: | MS_Analysis_Protocol.pdf |
| MS ID: | MS006817 |
| Analysis ID: | AN007121 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The mass spectrometer was operated in both positive and negative modes with a scan range 200-1500 m/z under 30,000 resolution at 200 m/z. Electrospray ionization (ESI) settings were: sheath gas 50, aux gas 13, spray voltage 3.5 kV, capillary temperature 263°C, S-lens RF 55, and heater temperature 425°C. MS/MS data acquisition settings were: microscans 2, resolution 15,000 at 200 m/z, maximum injection time 100 ms, and collision energies 20 and 30 eV. |
| Ion Mode: | UNSPECIFIED |
| Analysis Protocol File: | MS_Analysis_Protocol.pdf |
