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.

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Study IDST001289
Study TitleRegulated accumulation of desmosterol integrates macrophage lipid metabolism and inflammatory responses
Study SummaryTo 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
DepartmentMultiple
LaboratoryMultiple
Last NameFahy
First NameEoin
Address9500 Gilman, La Jolla, CA, 92093, USA
Emailefahy@ucsd.edu
Phone858-534-4076
Submit Date2019-12-17
PublicationsSpann 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 DetailMS
Release Date2020-01-22
Release Version1
Eoin Fahy Eoin Fahy
https://dx.doi.org/10.21228/M83M5X
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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
SA0936808_4LDLR-KO High fat
SA0936818_2LDLR-KO High fat
SA0936828_1LDLR-KO High fat
SA0936838_5LDLR-KO High fat
SA0936848_3LDLR-KO High fat
SA0936858_6LDLR-KO High fat
SA0936868_9LDLR-KO High fat
SA0936878_8LDLR-KO High fat
SA0936888_7LDLR-KO High fat
SA0936897_5LDLR-KO Normal
SA0936907_4LDLR-KO Normal
SA0936917_6LDLR-KO Normal
SA0936927_3LDLR-KO Normal
SA0936937_9LDLR-KO Normal
SA0936947_2LDLR-KO Normal
SA0936957_8LDLR-KO Normal
SA0936967_7LDLR-KO Normal
SA0936977_1LDLR-KO Normal
SA0936986_4Wild type High fat
SA0936996_3Wild type High fat
SA0937006_1Wild type High fat
SA0937016_5Wild type High fat
SA0937026_2Wild type High fat
SA0937036_9Wild type High fat
SA0937046_6Wild type High fat
SA0937056_8Wild type High fat
SA0937066_7Wild type High fat
SA0937075_4Wild type Normal
SA0937085_2Wild type Normal
SA0937095_5Wild type Normal
SA0937105_3Wild type Normal
SA0937115_8Wild type Normal
SA0937125_1Wild type Normal
SA0937135_9Wild type Normal
SA0937145_7Wild type Normal
SA0937155_6Wild 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 Several
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
Chromatography Type: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
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