Project description:Homozygous K107R mutation of PPARg in mice alters the expression of its downstream target genes and increases insulin sensitivity but not adiposity.
Project description:Adipose tissue gene expression was profiled from perigonadal adipose tissue of mice whose body mass and insulin sensitivity varied as a function of diet, gender, monogenic mutations and insulin sensitizing therapies. Mice were 22-24 weeks of age and sacrificed between 2-3 hours into the light portion of a 12/12 dark-light cycle. The goal was to identify transcripts whose adipose tissue expression is correlated with adiposity, insulin sensitivity and other measures of metabolic function. Keywords: population
Project description:Adipose tissue gene expression was profiled from perigonadal adipose tissue of mice whose body mass and insulin sensitivity varied as a function of diet, gender, monogenic mutations and insulin sensitizing therapies. Mice were 22-24 weeks of age and sacrificed between 2-3 hours into the light portion of a 12/12 dark-light cycle. The goal was to identify transcripts whose adipose tissue expression is correlated with adiposity, insulin sensitivity and other measures of metabolic function. Experiment Overall Design: Mice on the C57BL/6J strain were housed in groups of 3-5 and fed ad libitum. 2-4 days prior to collection of adipose tissue fasting blood glucose and serum insulin concentrations were measured. Mice were sacrificed 2-3 hours into the light cycle. RNA was extracted from perigonadal adipose tissue and used to generate labed cRNA for hybridization to Affymetrix Mu74Av2 microarrays.
Project description:Breast cancer is genetically heterogeneous, and recent studies have underlined a prominent contribution of epigenetics to the development of this disease. To uncover new synthetic lethalities with known breast cancer oncogenes, we screened an epigenome-focused short hairpin RNA library on a panel of engineered breast epithelial cell lines. Here we report a selective interaction between the NOTCH1 signaling pathway and the SUMOylation cascade. Knockdown of the E2-conjugating enzyme UBC9 (UBE2I) as well as inhibition of the E1-activating complex SAE1/UBA2 using ginkgolic acid impairs the growth of NOTCH1-activated breast epithelial cells. We show that upon inhibition of SUMOylation NOTCH1-activated cells proceed slower through the cell cycle and ultimately enter apoptosis. Mechanistically, activation of NOTCH1 signaling depletes the pool of unconjugated small ubiquitin-like modifier 1 (SUMO1) and SUMO2/3 leading to increased sensitivity to perturbation of the SUMOylation cascade. Depletion of unconjugated SUMO correlates with sensitivity to inhibition of SUMOylation also in patient-derived breast cancer cell lines with constitutive NOTCH pathway activation. Our investigation suggests that SUMOylation cascade inhibitors should be further explored as targeted treatment for NOTCH-driven breast cancer. We treated MCF10A and NOTCH1 cells with either DMSO or ginkgolic acid 30 uM for 3 days. Two replicates have been analysed for each condition.
Project description:In adipocyte-specific knockout mice (Bcl6AKO), we found that Bcl6 deletion results in strikingly increased inguinal but not perigonadal adipocyte size and tissue mass in addition to marked insulin sensitivity. Genome-wide DNA binding and RNA expression analyses revealed that BCL6 controls gene networks involved in cell growth and fatty acid biosynthesis. Thus, our studies identify BCL6 as a negative regulator of subcutaneous adipose tissue expansion and metabolic health.
Project description:In adipocyte-specific knockout mice (Bcl6AKO), we found that Bcl6 deletion results in strikingly increased inguinal but not perigonadal adipocyte size and tissue mass in addition to marked insulin sensitivity. Genome-wide DNA binding and RNA expression analyses revealed that BCL6 controls gene networks involved in cell growth and fatty acid biosynthesis. Thus, our studies identify BCL6 as a negative regulator of subcutaneous adipose tissue expansion and metabolic health.
Project description:Adipose tissue dysfunction in obese humans is associated with disrupted metabolic homeostasis, insulin resistance, and type 2 diabetes mellitus (T2DM). In a mouse model that has preserved insulin sensitivity despite increased adiposity, we used unbiased three-dimensional integration of proteome profiles, metabolic profiles, and gene regulatory networks to understand adipose tissue proteome-wide changes and their metabolic implications. Multiple-dimensional liquid chromatography tandem mass spectrometry and extended multiplexing TMT labeling (24 biological samples) was used to analyze proteomes of epididymal adipose tissues isolated from wildtype (Csf2+/+) and GM-CSF driven dendritic cell deficient (Csf2-/-) mice that were fed low fat, high fat, or high fat plus cholesterol diets for 8 weeks. The peripheral metabolic health (as measured by body weight, adiposity, plasma fasting glucose, insulin, triglycerides, phospholipids, total cholesterol levels, and glucose and insulin tolerance tests) deteriorated with diet for both genotypes, while mice lacking Csf2 were protected from insulin resistance. Regardless of diet, 30, mostly mitochondrial metabolism proteins participating in amino acid and branched chain amino acid pathways were altered, between Csf2-/- and Csf2+/+ mice. Tissue DHTKD1 levels were >4-fold upregulated and plasma 2-aminoadipoate (2-AA) levels were >2 fold reduced in Csf2-/- mice. GM-CSF driven dendritic cells play a detrimental role in insulin sensitivity via lysine metabolism involving Dhtkd1/2-AA axis.