Unknown,Transcriptomics,Genomics,Proteomics

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GReD_Prostate_LXR

ABSTRACT: Lxr-/- mice and WT mice were fed with a western diet and a control diet; Transcriptomic analysis of total RNA samples originated from latero-dorsal prostates was performed as described below between the four conditions: Wild type Mice +/+ on normal diet, Wild type Mice +/+ on western diet, lxr KO Mice-/- on normal diet, lxr KO Mice-/- on western diet. 4 samples will be pooled for each condition: WT +/+ base diet WT +/+ western diet, LXR -/- base diet LXR -/- western diet. Analysis of diet effects: 1. On WT mice: WT Mice +/+ normal diet vs WT Mice +/+ western diet. Wild type base diet RNA pool will be compared to Wild type western diet RNA pool by performing a direct comparison on an Agilent array using the 2-color approach. Dye flips will be prepared for a total of 2 hybridizations. 2. On KO mice: LXR mice -/- base diet vs LXR mice -/- western diet. LXR KO base diet RNA pool will be compared to LXR KO western diet RNA pool by performing a direct comparison on an Agilent array using the 2-color approach. Dye flips will be prepared for a total of 2 hybridizations. Analysis of genotype effects: 1. Under western diet condition: WT +/+ western diet vs LXR -/- western diet. Wild type western diet RNA pool will be compared to LXR KO western diet RNA pool by performing a direct comparison on an Agilent array using the 2-color approach. Dye flips will be prepared for a total of 2 hybridizations. 2. Under base diet condition: WT +/+ base diet vs LXR -/- base diet Wild type base diet RNA pool will be compared to LXR KO base diet RNA pool by performing a direct comparison on an Agilent array using the 2-color approach. Dye flips will be prepared for a total of 2 hybridizations.

ORGANISM(S): Mus musculus

SUBMITTER: Silvere Baron

PROVIDER: E-MTAB-546 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description: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.

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GReD_Prostate_LXR

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