Summary of Study ST002093
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 PR001329. The data can be accessed directly via it's Project DOI: 10.21228/M8XM74 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST002093 |
| Study Title | Lipidomics of High Fat vs Control Mice |
| Study Type | Untargeted Lipidomics |
| Study Summary | Obesity exacerbates inflammation upon lung injury; however, the mechanisms by which obesity primes pulmonary dysregulation prior to injury are not well studied. Notably, little is known about how obesity dysregulates pulmonary polyunsaturated fatty acid (PUFA) metabolism that is central to inflammation initiation and resolution. Herein, we first show with mass spectrometry that a high fat diet (HFD) administered to C57BL/6J mice increases the relative abundance of pulmonary PUFA-containing triglycerides and the concentration of PUFA-derived oxylipins, independent of an increase in total pulmonary PUFAs. Experiments with a genetic model of obesity did not recapitulate the effects of the HFD on the pulmonary oxylipin signature, suggesting a diet-driven effect. Subsequent pulmonary next-generation RNA sequencing showed complex and unique transcriptional regulation with a HFD. The HFD increased pathways related to glycerophospholipid metabolism, innate immunity, and inflammation including an elevation in B cell differentiation and signaling. Finally, computational integration of lipidomic with transcriptomic data revealed novel networks with the HFD between glycerophospholipid metabolism and B cell receptor signaling with specific oxylipins. Collectively, these data show obesity dysregulates pulmonary PUFA metabolism prior to lung injury, which may be a mechanism by which obesity primes the lungs to respond poorly upon infectious and/or inflammatory challenges. |
| Institute | University of North Carolina at Chapel Hill |
| Department | Chemistry |
| Laboratory | MS Core Laboratory |
| Last Name | Weatherspoon |
| First Name | Emily |
| Address | 131 South Rd |
| emdiane@email.unc.edu | |
| Phone | 7042453664 |
| Submit Date | 2022-02-23 |
| Num Groups | 2 |
| Total Subjects | 3 |
| Num Males | 6 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Waters) |
| Analysis Type Detail | LC-MS |
| Release Date | 2022-03-15 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001329 |
| Project DOI: | doi: 10.21228/M8XM74 |
| Project Title: | Lipidomics of High Fat vs Control Mice |
| Project Type: | Untargeted LC-MS/MS Lipidomics |
| Project Summary: | Obesity exacerbates inflammation upon lung injury; however, the mechanisms by which obesity primes pulmonary dysregulation prior to injury are not well studied. Notably, little is known about how obesity dysregulates pulmonary polyunsaturated fatty acid (PUFA) metabolism that is central to inflammation initiation and resolution. Herein, we first show with mass spectrometry that a high fat diet (HFD) administered to C57BL/6J mice increases the relative abundance of pulmonary PUFA-containing triglycerides and the concentration of PUFA-derived oxylipins, independent of an increase in total pulmonary PUFAs. Experiments with a genetic model of obesity did not recapitulate the effects of the HFD on the pulmonary oxylipin signature, suggesting a diet-driven effect. Subsequent pulmonary next-generation RNA sequencing showed complex and unique transcriptional regulation with a HFD. The HFD increased pathways related to glycerophospholipid metabolism, innate immunity, and inflammation including an elevation in B cell differentiation and signaling. Finally, computational integration of lipidomic with transcriptomic data revealed novel networks with the HFD between glycerophospholipid metabolism and B cell receptor signaling with specific oxylipins. Collectively, these data show obesity dysregulates pulmonary PUFA metabolism prior to lung injury, which may be a mechanism by which obesity primes the lungs to respond poorly upon infectious and/or inflammatory challenges. |
| Institute: | University of North Carolina at Chapel Hill |
| Department: | Chemistry |
| Laboratory: | MS Core Laboratory |
| Last Name: | Weatherspoon |
| First Name: | Emily |
| Address: | 131 South Rd |
| Email: | emdiane@email.unc.edu |
| Phone: | 7042453664 |
Subject:
| Subject ID: | SU002177 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 21 weeks |
| Gender: | Male |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA200928 | CON-L-8RL | Control |
| SA200929 | CON-R-7RN | Control |
| SA200930 | CON-L-8LN | Control |
| SA200931 | Con-L-7RN | Control |
| SA200932 | CON-L-7NW | Control |
| SA200933 | CON-R-8RL | Control |
| SA200934 | HF-L-3MN | High Fat |
| SA200935 | HF-L-5L | High Fat |
| SA200936 | HF-R-4L | High Fat |
| SA200937 | HF-L-4RL | High Fat |
| SA200938 | HF-L-4R | High Fat |
| SA200939 | HF-L-4L | High Fat |
| SA200940 | HF-R-4R | High Fat |
| Showing results 1 to 13 of 13 |
Collection:
| Collection ID: | CO002170 |
| Collection Summary: | C57BL/6J male mice were purchased from Jackson Laboratories. At 6 weeks of age, mice were fed lean control (10% kcal from fat) or high-fat diet (60% kcal from fat) for 15 weeks (Research Diets, New Brunswick, NJ). Ob/ob mice, a genetic model of obesity driven by leptin deficiency, were purchased from Jackson laboratories at age 10 weeks and were fed a normal chow diet for an additional 5 weeks. |
| Sample Type: | Lung |
Treatment:
| Treatment ID: | TR002189 |
| Treatment Summary: | C57BL/6J male mice were purchased from Jackson Laboratories. At 6 weeks of age, mice were fed lean control (10% kcal from fat) or high-fat diet (60% kcal from fat) for 15 weeks (Research Diets, New Brunswick, NJ). Ob/ob mice, a genetic model of obesity driven by leptin deficiency, were purchased from Jackson laboratories at age 10 weeks and were fed a normal chow diet for an additional 5 weeks. |
Sample Preparation:
| Sampleprep ID: | SP002183 |
| Sampleprep Summary: | Mouse lungs were weighed into Eppendorf tubes. The lung tissues were mashed using a clean metal spatula prior to extraction. The samples were extracted using a liquid-liquid partition with water (250 µL), methanol (300 µL), and methyl tert-butyl ether (MTBE). Avanti’s deuterated lipid mix, Equisplash, was used as an internal standard. The deuterated mix was spiked into the methanol at 1.5 µg/mL and used for extraction. The extracts were centrifuged at 20,000 rcf for 10 minutes to facilitate phase separation. The top layer was removed, dried, and reconstituted in 150 µL of isopropanol for analysis. |
| Processing Storage Conditions: | -80℃ |
| Sample Resuspension: | 150 µL of IPA |
Combined analysis:
| Analysis ID | AN003419 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity I-Class |
| Column | Waters Acquity BEH C18 (100 x 2mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | POSITIVE |
| Units | Peak Area |
Chromatography:
| Chromatography ID: | CH002529 |
| Instrument Name: | Waters Acquity I-Class |
| Column Name: | Waters Acquity BEH C18 (100 x 2mm,1.7um) |
| Column Temperature: | 45 |
| Flow Gradient: | Starting composition was 32% B, which increased to 40% B at 1 min (held until 1.5 min) then 45% B at 4 minutes. This was increased to 50% B at 5 min, 60% B at 8 min, 70% B at 11 min, and 80% B at 14 min (held until 16 min). At 16 min the composition switched back to starting conditions (32% B) and was held for 4 min to re-equilibrate the column. |
| Flow Rate: | 0.2 mL/min |
| Injection Temperature: | 10 |
| Solvent A: | 40% water/60% acetonitrile; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003184 |
| Analysis ID: | AN003419 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Samples were analyzed with positive/negative ionization switching. 60,000 resolution. Top 5 DDA. |
| Ion Mode: | POSITIVE |
| Collision Energy: | 25, 35, 45 V stepped collision energy |
| Fragmentation Method: | HCD |
| Ionization: | HESI |
| Mass Accuracy: | 5 ppm |
