Project description:SP1-regulated transcriptome: Gene expression microarray following SP1 RNAi to define the SP1-regulated transcriptome in human in vitro differentiated adipocyte. Glutamine-regulated transcriptome: Gene expression microarray in human in vitro differentiated adipocyte incubated in high (10 mM) and low (0.5 mM) concentrations of glutamine.
Project description:Mapping of O-GlcNAcylated proteome along the genome by chromatin immunoprecipitation followed by DNA sequencing (ChIP-seq) in in vitro differentiated human adipocytes incubated with low (0.5 mM) or high (10 mM) glutamine.
Project description:Obesity-driven pathological expansion of white adipose tissue (WAT) is a key driver of endothelial dysfunction. Contrary to this paradigm, early vascular alterations associated with over nutrition also exacerbate AT dysfunction. To dissect this complex cause and consequence relationship, here we perform a single-cell transcriptomics screen to generate a detailed landscape of endothelial heterogeneity and vascular alterations in murine model of obesity. Given the differences in ontogeny and function of distinct WAT depots, we demarcate key differences in subcutaneous and visceral WAT vasculature. In addition to descriptive taxonomy, we perform in-depth validation and characterization of our in silico data. We identify a sWAT specific fenestrated endothelial cell subtype, which is drastically reduced in obese conditions. This reduction was associated with a decrease in VEGFA expressing perivascular cells. The novel endothelial subtypes provide a basis for future research and new directions for therapeutic interventions.
Project description:Glutamine plays a key role in cellular metabolism and tissue homeostasis. In obesity, circulating glutamine levels are decreased, accompanied by impaired bone homeostasis and increased fracture risk. While glutamine supplementation in diet has shown metabolic benefits, the mechanisms behind its effects on bone and fat metabolism are not fully understood. This study investigates whether glutamine supplementation may delay the negative impact of obesity on bone and fat metabolism in mice fed a high-fat diet (HFD). Two-month-long dietary intervention in C57BL/6J male mice showed that glutamine supplementantion reduced body weight gain, fat mass, and white adipose tissue (WAT) weight, along with improved glucose tolerance compared to HFD-fed mice. In peripheral WAT, glutamine supplementation improved WAT funcion, as indicated by decreased adipocyte hypertrophy and inflammatory status. Furthermore, glutamine reduced the obesity-driven hyper-metabolic phenotype in primary adipose-derived mesenchymal stem cells by shifting the cells toward a quiescent metabolic state. In bone, glutamine supplementation improved bone quality and microarchitecture, along with reduced bone marrow adiposity and decreased bone resorption in glutamine-treated mice compared to HFD-fed mice. These changes were confirmed by decreased adipogenic and increased osteogenic potential of bone marrow stromal cells isolated from glutamine-treated mice. This was further supported by enhanced glutamine turnover, which maintained the stemness of the cells and reduced the inflammatory status induced by obesity, The importance of glutamine metabolism was also confirmed in human stem cells, showing a sex-specific pattern of glutaminolysis. Thus, our findings demonstrate that glutamine supplementation in obesity improves metabolic health and bone integrity at both the organ and cellular levels, highlighting its potential as a therapeutic strategy for preventing obesity-related metabolic and bone diseases.
Project description:Obesity is associated with chronic low-grade white adipose tissue (WAT) inflammation that can contribute to the development of insulin resistance in mammals. Previous studies have identified interleukin (IL)-12 as a critical upstream regulator of WAT inflammation and metabolic dysfunction during obesity, however, the cell types and mechanisms that initiate WAT IL-12 production remain unclear. Analysis of mouse and human WAT single cell transcriptomic datasets, IL-12 reporter mice, and IL-12p70 protein levels by ELISA identified activated conventional type 1 dendritic cells (cDC1s) as the cellular source of WAT IL-12 during diet-induced obesity. cDC1s were required for the development of obesity-associated inflammation by increasing group 1 innate lymphocyte interferon (IFN)-γ production and inflammatory macrophage accumulation. Inducible depletion of cDC1s increased WAT insulin sensitivity and systemic glucose tolerance during diet-induced obesity. Endocytosis of apoptotic bodies containing self-DNA by WAT cDC1 drove STING-dependent IL-12 production. Together, these results suggest that WAT cDC1s act as critical regulators of adipose tissue inflammation and metabolic homeostasis during obesity.
Project description:Obesity is a significant risk factor for diabetes, cardiovascular diseases, and certain cancers, and manifests as excessive fat accumulation. The browning of white adipose tissue (WAT) represents one of the most promising strategies for preventing and treating obesity and metabolic diseases. To date, an increasing number of studies have focused on key molecular mechanisms regulating fat thermogenesis, laying the foundation for effective intervention strategies.We analyzed the differentially expressed proteins in white adipose tissue of the inguinal region using mass spectrometry.