Summary of Study ST002800
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 PR001747. The data can be accessed directly via it's Project DOI: 10.21228/M8WH9J 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 | ST002800 |
| Study Title | Hepatic oxylipin profiles in mouse models of Wilson disease: new insights into early hepatic manifestations. |
| Study Summary | Hepatic inflammation is commonly identified in Wilson disease (WD), a genetic disease of hepatic and brain copper accumulation. Copper accumulation is associated with increased reactive oxygen species and activation of the non-enzymatic oxidation of membrane-bound polyunsaturated fatty acids (PUFA), with impairment of cellular structures and function. Products of PUFA oxidation are collectively known as oxylipins (OXL), which can also be produced via enzymatic pathways including lipoxygenases (LOXs), cyclooxygenases (COXs), and cytochrome P450 monooxygenases (CYPs). These bioactive lipids modulate hepatic inflammation. We aimed to examine hepatic OXLs profile at early stages of WD in mouse model of Wilson disease, the toxic milk from The Jackson Laboratory (tx-j) compared to mice with normal copper metabolism (C3H). Targeted lipidomic profiling of OXLs was performed by ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) in livers from 16 weeks old male and female mice. Hepatic OXL profiles were altered, with higher levels of PUFA alcohols, diols, and ketones. Markers of oxidative stress, 9-HETE and 9-HEPE were increased in tx-j mice. Hepatic prostaglandin and thromboxane levels in the COX pathway were increased in tx-j mice. tx-j showed altered PUFA-epoxides, suggesting altered CYP(s) activities. Our findings suggest that both non-enzymatic ROS-dependent and enzymatic PUFAs oxidation via COX and LOX pathways are associated with early stages liver disease in WD. It also indicates altered CYPs activities in animal model of WD. These novel pathways could be the target for future therapies. |
| Institute | University of California, Davis |
| Last Name | Sarode |
| First Name | Gaurav |
| Address | Genome & Biomedical Sciences Facility, 451 E. Health Sciences Dr., Davis, CA, 95616, USA |
| gsarode@ucdavis.edu | |
| Phone | 530752-6715 |
| Submit Date | 2023-07-18 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | wiff |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-01-18 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001747 |
| Project DOI: | doi: 10.21228/M8WH9J |
| Project Title: | Hepatic oxylipin profiles in mouse models of Wilson disease: new insights into early hepatic manifestations. |
| Project Summary: | Hepatic inflammation is commonly identified in Wilson disease (WD), a genetic disease of hepatic and brain copper accumulation. Copper accumulation is associated with increased reactive oxygen species and activation of the non-enzymatic oxidation of membrane-bound polyunsaturated fatty acids (PUFA), with impairment of cellular structures and function. Products of PUFA oxidation are collectively known as oxylipins (OXL), which can also be produced via enzymatic pathways including lipoxygenases (LOXs), cyclooxygenases (COXs), and cytochrome P450 monooxygenases (CYPs). These bioactive lipids modulate hepatic inflammation. We aimed to examine hepatic OXLs profile at early stages of WD in mouse model of Wilson disease, the toxic milk from The Jackson Laboratory (tx-j) compared to mice with normal copper metabolism (C3H). Targeted lipidomic profiling of OXLs was performed by ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) in livers from 16 weeks old male and female mice. Hepatic OXL profiles were altered, with higher levels of PUFA alcohols, diols, and ketones. Markers of oxidative stress, 9-HETE and 9-HEPE were increased in tx-j mice. Hepatic prostaglandin and thromboxane levels in the COX pathway were increased in tx-j mice. tx-j showed altered PUFA-epoxides, suggesting altered CYP(s) activities. Our findings suggest that both non-enzymatic ROS-dependent and enzymatic PUFAs oxidation via COX and LOX pathways are associated with early stages liver disease in WD. It also indicates altered CYPs activities in animal model of WD. These novel pathways could be the target for future therapies. |
| Institute: | University of California, Davis |
| Department: | Internal Medicine |
| Laboratory: | Medici's Lab |
| Last Name: | Sarode |
| First Name: | Gaurav |
| Address: | Genome & Biomedical Sciences Facility, 451 E. Health Sciences Dr., Davis, CA, 95616, USA |
| Email: | gsarode@ucdavis.edu |
| Phone: | 530752-6715 |
Subject:
| Subject ID: | SU002907 |
| 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 | Treatment |
|---|---|---|
| SA300730 | 930 | C3H 5001 |
| SA300731 | 928 | C3H 5001 |
| SA300732 | 926 | C3H 5001 |
| SA300733 | 900 | C3H 5001 |
| SA300734 | 927 | C3H 5001 |
| SA300735 | 929 | C3H 5001 |
| SA300736 | 906 | C3H 5001 |
| SA300737 | 901 | C3H 5001 |
| SA300738 | 903 | C3H 5001 |
| SA300739 | 902 | C3H 5001 |
| SA300740 | 905 | C3H 5001 |
| SA300741 | 904 | C3H 5001 |
| SA300742 | 922 | tx-J 5001 |
| SA300743 | 921 | tx-J 5001 |
| SA300744 | 923 | tx-J 5001 |
| SA300745 | 924 | tx-J 5001 |
| SA300746 | 920 | tx-J 5001 |
| SA300747 | 925 | tx-J 5001 |
| SA300748 | 916 | tx-J 5001 |
| SA300749 | 911 | tx-J 5001 |
| SA300750 | 910 | tx-J 5001 |
| SA300751 | 909 | tx-J 5001 |
| SA300752 | 908 | tx-J 5001 |
| SA300753 | 912 | tx-J 5001 |
| SA300754 | 913 | tx-J 5001 |
| SA300755 | 918 | tx-J 5001 |
| SA300756 | 917 | tx-J 5001 |
| SA300757 | 915 | tx-J 5001 |
| SA300758 | 914 | tx-J 5001 |
| SA300759 | 919 | tx-J 5001 |
| Showing results 1 to 30 of 30 |
Collection:
| Collection ID: | CO002900 |
| Collection Summary: | The liver was isolated. All samples were stored at -80°C until further analysis. |
| Sample Type: | Liver |
Treatment:
| Treatment ID: | TR002916 |
| Treatment Summary: | NA |
Sample Preparation:
| Sampleprep ID: | SP002913 |
| Sampleprep Summary: | Weigh 1-15 mg tissue aliquots on dry ice, record exact weights, place in 2-mL eppendorf tubes labeled on cap, keep frozen Add 10 µL anti-oxidant soln, (0.2 mg/ml solution BHT/EDTA in 1:1 meoh:water) Add 10 uL of surrogate standards Add 500 uL of cold methanol as extraction solvent and stainless steel grinding balls Homogenize using GenoGrinder 2x30 sec, centrifuge, transfer supernatant to a 1.5-mL eppendorf tube (Cryo-label on side) containing 10 µL 20% glycerol solution in MeOH Add a second aliquot of 500 uL of cold methanol to centrifugation pellet Homogenize using GenoGrinder 2x30 sec, centrifuge, combine this second supernatant with the first one in the 1.5-mL eppendorf tube Transfer vials to Speed-vac and evaporate to dryness or transfer to 96 well plate and Gene-Vac to dryness. Assure samples are dry before removing from evaporation process. Reconstitute samples for LC-MS in PHAU/CUDA 50 nM in methanol/ACN 50:50, vortex for 10 sec, then sonicate for 5 min. Set rack of samples on wet ice for 15min Centrifuge for 3 min at highest speed, then transfer supernatant to glass insert in amber HPLC vial / 96 well plate Store at -20°C until LCMS analysis |
Combined analysis:
| Analysis ID | AN004556 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Shimadzu Nexera X2 |
| Column | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Triple quadrupole |
| MS instrument name | ABI Sciex 6500 QTrap |
| Ion Mode | UNSPECIFIED |
| Units | nM |
Chromatography:
| Chromatography ID: | CH003423 |
| Instrument Name: | Shimadzu Nexera X2 |
| Column Name: | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
| Column Temperature: | 65°C |
| Flow Gradient: | 75% ACN:IPA 90:10 (v/v) with 0.1% Acetic acid |
| Flow Rate: | 0.6 mL/min |
| Solvent A: | 100% water; 0.1% acetic acid |
| Solvent B: | 90% acetonitrile/ 10% isopropanol |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004303 |
| Analysis ID: | AN004556 |
| Instrument Name: | ABI Sciex 6500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | LC/MS on Thermo Scientific Vanquish UPLC/Sciex Qtrap with targeted MRM method (developed by Newman lab, optimized for Fiehn lab setup). See protocol file. |
| Ion Mode: | UNSPECIFIED |
| Analysis Protocol File: | Oxy-EC_analysis_protocol.pdf |
