Summary of Study ST001289
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 PR000871. The data can be accessed directly via it's Project DOI: 10.21228/M83M5X This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001289 |
| Study Title | Regulated accumulation of desmosterol integrates macrophage lipid metabolism and inflammatory responses |
| Study Summary | To investigate the relationship between hypercholesterolemia, foam cell formation and inflammation, we performed lipidomic and transcriptomic analyses of elicited peritoneal macrophages in wild type (WT) or LDL receptor knockout (LDLR KO) mice fed either a normal cholesterol, normal fat (NCNF) diet or a high cholesterol, high fat (HCHF) 'Western' style diet. The combination of the LDLR KO genotype and the HCHF diet results in the formation of macrophage foam cells in the elicited peritoneal macrophage population. Analysis of macrophages from the above four experimental groups revealed massive reprogramming of the lipidome in response to both diet and genotype. These studies confirmed and extended prior knowledge regarding the roles of SREBP and LXR signaling in cholesterol and fatty acid homeostasis. Unexpectedly, peritoneal macrophage foam cells exhibited a strongly 'deactivated' phenotype, with marked suppression of pro-inflammatory mediators that are normally characteristic of the inflammatory responses associated with atherosclerotic lesions. Many of these changes in gene expression and lipid metabolism appear to be related to the paradoxical accumulation of high levels of desmosterol, the last intermediate in the Bloch pathway of cholesterol biosynthesis. WT or LDLR KO mice were fed either a NCNF diet or a HCHF diet for twelve weeks to establish four experimental groups (WT-NCNF diet, WT-HCHF diet, KO-NCNF diet, and KO-HCHF diet). As expected, the combination of the HCHF diet and LDLR KO genotype resulted in a synergistic effect on serum lipid levels. Elicited peritoneal macrophages (92-96% F4/80-positive) were immediately prepared for analysis, thereby preserving in vivo gene expression and lipid profiles. Macrophages derived from LDLR KO mice fed the HCHF diet contained nearly four-fold more total cholesterol than cells from WT mice fed the same diet. Quantitative analysis of 245 lipid species revealed significant changes in nearly all major lipid classes. Using a two-way ANOVA model, we found that 176 (72%) of the lipids analyzed were significantly affected by the HCHF diet, 133 (54%) by the LDLR KO genotype, and 114 (46%) by interactions between the HCHF diet and LDLR KO genotype. Many of the observed interactions (60%) were synergistic. |
| Institute | LIPID MAPS |
| Department | Multiple |
| Laboratory | Multiple |
| Last Name | Fahy |
| First Name | Eoin |
| Address | 9500 Gilman, La Jolla, CA, 92093, USA |
| efahy@ucsd.edu | |
| Phone | 858-534-4076 |
| Submit Date | 2019-12-17 |
| Publications | Spann NJ, Garmire LX, McDonald JG, Myers DS, Milne SB, Shibata N, Reichart D, Fox JN, Shaked I, Heudobler D, Raetz CR, Wang EW, Kelly SL, Sullards MC, Murphy RC, Merrill AH Jr, Brown HA, Dennis EA, Li AC, Ley K, Tsimikas S, Fahy E, Subramaniam S, Quehenberger O, Russell DW, Glass CK. Regulated accumulation of desmosterol integrates macrophage lipid metabolism and inflammatory responses. Cell. 2012 Sep 28;151(1):138-52. doi: 10.1016/j.cell.2012.06.054. PMID: 23021221; PMCID: PMC3464914. |
| Analysis Type Detail | GC-MS/LC-MS |
| Release Date | 2020-01-22 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR000871 |
| Project DOI: | doi: 10.21228/M83M5X |
| Project Title: | Regulated accumulation of desmosterol integrates macrophage lipid metabolism and inflammatory responses |
| Project Summary: | Inflammation and macrophage foam cells are characteristic features of atherosclerotic lesions, but the mechanisms linking cholesterol accumulation to inflammation and LXR-dependent response pathways are poorly understood. To investigate this relationship, we utilized lipidomic and transcriptomic methods to evaluate the effect of diet and LDL receptor genotype on macrophage foam cell formation within the peritoneal cavities of mice. Foam cell formation was associated with significant changes in hundreds of lipid species and unexpected suppression, rather than activation, of inflammatory gene expression. We provide evidence that regulated accumulation of desmosterol underlies many of the homeostatic responses observed in macrophage foam cells, including activation of LXR target genes, inhibition of SREBP target genes, selective reprogramming of fatty acid metabolism and suppression of inflammatory response genes. These observations suggest that macrophage activation in atherosclerotic lesions results from extrinsic, pro-inflammatory signals generated within the artery wall that suppress homeostatic and anti-inflammatory functions of desmosterol. |
| Institute: | University of California, San Diego |
| Department: | Bioengineering |
| Last Name: | Fahy |
| First Name: | Eoin |
| Address: | 9500 Gilman, La Jolla, CA, 92093, USA |
| Email: | efahy@ucsd.edu |
| Phone: | 858-534-4076 |
Subject:
| Subject ID: | SU001361 |
| 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 | Genotype | Diet |
|---|---|---|---|
| SA093680 | 8_4 | LDLR-KO | High fat |
| SA093681 | 8_2 | LDLR-KO | High fat |
| SA093682 | 8_1 | LDLR-KO | High fat |
| SA093683 | 8_5 | LDLR-KO | High fat |
| SA093684 | 8_3 | LDLR-KO | High fat |
| SA093685 | 8_6 | LDLR-KO | High fat |
| SA093686 | 8_9 | LDLR-KO | High fat |
| SA093687 | 8_8 | LDLR-KO | High fat |
| SA093688 | 8_7 | LDLR-KO | High fat |
| SA093689 | 7_5 | LDLR-KO | Normal |
| SA093690 | 7_4 | LDLR-KO | Normal |
| SA093691 | 7_6 | LDLR-KO | Normal |
| SA093692 | 7_3 | LDLR-KO | Normal |
| SA093693 | 7_9 | LDLR-KO | Normal |
| SA093694 | 7_2 | LDLR-KO | Normal |
| SA093695 | 7_8 | LDLR-KO | Normal |
| SA093696 | 7_7 | LDLR-KO | Normal |
| SA093697 | 7_1 | LDLR-KO | Normal |
| SA093698 | 6_4 | Wild type | High fat |
| SA093699 | 6_3 | Wild type | High fat |
| SA093700 | 6_1 | Wild type | High fat |
| SA093701 | 6_5 | Wild type | High fat |
| SA093702 | 6_2 | Wild type | High fat |
| SA093703 | 6_9 | Wild type | High fat |
| SA093704 | 6_6 | Wild type | High fat |
| SA093705 | 6_8 | Wild type | High fat |
| SA093706 | 6_7 | Wild type | High fat |
| SA093707 | 5_4 | Wild type | Normal |
| SA093708 | 5_2 | Wild type | Normal |
| SA093709 | 5_5 | Wild type | Normal |
| SA093710 | 5_3 | Wild type | Normal |
| SA093711 | 5_8 | Wild type | Normal |
| SA093712 | 5_1 | Wild type | Normal |
| SA093713 | 5_9 | Wild type | Normal |
| SA093714 | 5_7 | Wild type | Normal |
| SA093715 | 5_6 | Wild type | Normal |
| Showing results 1 to 36 of 36 |
Collection:
| Collection ID: | CO001355 |
| Collection Summary: | Male WT C57BL6 or LDL receptor knockout mice (Jackson Laboratory) were fed a normal cholesterol/normal fat (NCNF) diet or a high cholesterol/high fat (HCHF) diet (Harlan Teklad, catalog # 96121 including 21% milk fat and 1.25% cholesterol) for 12 weeks in an IACUC-approved animal facility. Cells were harvested from the peritoneal cavity 4 days after i.p. administration of 3% thioglycollate medium. Cells were immediately counted, aliquoted, pelleted and sent for cDNA microarray analysis and lipid measurement. For all other experiments primary cells were isolated from male 6–8 week-old C57BL6 mice (Charles River Laboratories). |
| Sample Type: | Macrophages |
Treatment:
| Treatment ID: | TR001376 |
| Treatment Summary: | Mouse thioglycollate-elicited macrophages and bone marrow derived macrophages were obtained and cultured as described previously (Heinz et al., 2010). For various ligand treatments, cells were in Phenol-red free RPMI-1640 (Invitrogen) supplemented with either 10% heat-inactivated FBS (Hyclone) or 10% lipid-deficient FBS (Hyclone) for 24hr before treatment, then treated with either 1µM GW3965, 0.25-20µM desmosterol, or 0.3-30µg/ml cholesterol for 12-24hr. 50µM melavonate and 10µM Mevastatin (Sigma) were added to media in desmosterol response experiments. For TLR repression studies, cells were cultured in medium containing Phenol-red free RPMI 1640 (Invitrogen) with either heat-inactivated FBS (Hyclone) or lipid-deficient FBS (Hyclone) for 24hr, then pretreated with either 20-30µM desmosterol, 50µg/ml cholesterol, 5µM triparanol, or 50-100µM 9Z-palmitoleic acid for 18hr. Cells were stimulated with Pam3CSK4 (300ng/ml), PolyI:C (50ng/ml), LPS (100ng/ml), TNFa (30ng/ml), or KLA (100ng/ml) for 6hr. See Extended Experimental Procedures for additional details at https://doi.org/10.1016/j.cell.2012.06.054 |
Sample Preparation:
| Sampleprep ID: | SP001369 |
| Sampleprep Summary: | The various extraction procedures and lipidomic analyses for each lipid class are described in the publication |
Combined analysis:
| Analysis ID | AN002142 |
|---|---|
| Analysis type | MS |
| Chromatography type | |
| Chromatography system | Several |
| Column | Several |
| MS Type | Other |
| MS instrument type | Several |
| MS instrument name | Several |
| Ion Mode | UNSPECIFIED |
| Units | pmol/1E6 cells |
Chromatography:
| Chromatography ID: | CH001568 |
| Chromatography Summary: | See publication for details publication |
| Chromatography Comments: | See publicationpublication for details |
| Instrument Name: | Several |
| Column Name: | Several |
MS:
| MS ID: | MS001994 |
| Analysis ID: | AN002142 |
| Instrument Name: | Several |
| Instrument Type: | Several |
| MS Type: | Other |
| MS Comments: | See publicationpublication for details |
| Ion Mode: | UNSPECIFIED |
