Project description:PIEZO1 is a mechanically-activated ion channel that contributes to flow sensing in vascular endothelium. Moreover, deletion of endothelial PIEZO1 was recently found to suppress activation of Notch1 target genes in hepatic microvascular endothelium. Here, because of the liver’s dominant role in lipid regulation, we set out to test the novel hypothesis that endothelial PIEZO1 regulates hepatic lipid homeostasis. We performed bulk RNA sequencing on PIEZO1-deleted mice exposed to chow and high fat diets. Our transcriptomics analysis reveal unexpected relevance to lipid and glucose homeostasis.
Project description:The study aims at understanding the global nature of the immune responses within the the lung tissue 28 days post infection with Mycobacterium tuberculosis. Comparing the whole transcriptome of infected lungs to that of an uninfected lungs revealed a plethora of immune mechanisms driven by various cytokines. IFN-γ, IL-6, IL-2, TNFα were the major cytokines observed. We observed significant differential expression of gene involved in the JAK-STAT and the MAPK signalling pathways. We observed an interplay of the immune regulatory genes and various non-immune genes controlling the metabolism, apoptosis, cell growth, post translational modifications etc.
Project description:The aim of the project is to investigate whether endothelial-restricted augmentation of IGF-1 signaling is sufficient to suppress atherogenesis. We generated RNA-seq data from mice with endothelial-restricted over-expression of human wildtype IGF-1R (hIGFREO/ApoE-/-) or a signaling defective K1003R mutant human IGF-1R (mIGFREO/ApoE-/-).
Project description:Non-alcoholic fatty liver disease (NAFLD) is a major public health burden and it covers a spectrum of diseases. NAFLD starts with the accumulation of lipid droplets (LDs) within hepatocytes (steatosis). Part of the challenge of studying the mechanistic processes involved in LD accumulation and their implications on the pathogenesis of human NAFLD is due to the available models. Investigating hepatic LDs in humans is challenging and relies on liver biopsies, meaning only cross-sectional data be obtained. On the other hand, LD patterns in in vitro models are poorly defined and rarely reported. Diacylgylcerol acyltransferase (DGAT)2 is one of two enzymes that carry out the final committed step in triacylglycerol (TAG) synthesis. It is unclear whether the enzymes are able to compensate for each other or whether they have distinct roles. It has been hypothesised that DGAT1 primarily utilises exogenous fatty acids and DGAT2 uses de novo-derived fatty acids. Given the important role of this enzyme in TAG synthesis and accumulation, the aims of this study are first to create a cellular model of intrahepatocellular TAG accumulation by manipulating nutritional substrates and to investigate intracellular metabolism in wildtype and DGAT2 knockout cells under these conditions. The experimental workflow for this study is as follows: Huh7 cells (either wild type or knockout) were grown in media containing 11 mM glucose and 2% human serum (HS) for seven days before additional sugars and fatty acids (FAs) were added for a further seven days. All treatments contained 11 mM glucose and 2% HS, either with 200 µM FAs (low fat low sugar; LFLS), 5.5 mM fructose + 200 µM FAs (low fat high sugar; LFHS) or 5.5 mM fructose + 800 µM FAs (high fat high sugar; HFHS). FA metabolism, lipid droplet characteristics and transcriptomic signatures were investigated.
Project description:Synthetic glucocorticoids are the most potent anti-inflammatory agents known and are widely used therapeutically. However, frequent therapeutic use is accompanied by development of severe side effects notably fat accumulation, hyperglycaemia, and hepatosteatosis. It is known that GC binds glucocorticoid response elements (GREs) in the genome to either enhance, or repress gene transcription. To understand the mechanism of gene regulation by GC in detail RNASeq is used here to study gene expression patterns in either wild or reverb alpha knockout mice at different time point during the day and night, with DEX treatment.
Project description:Renewables-based biotechnology depends on enzymes to degrade plant lignocellulose to simple sugars that are converted to fuels or high-value products. Identification and characterization of such lignocellulose degradative enzymes could be fast-tracked by availability of an enzyme activity measurement method that is fast, label-free, uses minimal resources and allows direct identification of generated products. We developed such a method by applying carbohydrate arrays coupled with MALDI-ToF mass spectrometry to identify reaction products of carbohydrate active enzymes (CAZymes) of the filamentous fungus Aspergillus niger. We describe the production and characterization of plant polysaccharide-derived oligosaccharides and their attachment to hydrophobic self-assembling monolayers on a gold target. We verify effectiveness of this array for detecting exo- and endo-acting glycoside hydrolase activity using commercial enzymes, and demonstrate how this platform is suitable for detection of enzyme activity in relevant biological samples, the culture filtrate of A. niger grown on wheat straw. In conclusion, this versatile method is broadly applicable in screening and characterisation of activity of CAZymes, such as fungal enzymes for plant lignocellulose degradation with relevance to biotechnological applications as biofuel production, the food and animal feed industry.
Project description:To investigate if chronically elevated circulating kynurenic acid (KYNA) and reduced quinolinic acid (QUIN) levels affect whole-body energy metabolism, we used a knockout-first gene targeting strategy to eliminate Kmo gene expression in mice. We conducted a series of metabolic assessments on male and female mice fed either a chow or high-fat diet (HFD) and performed RNA-sequencing on the inguinal white adipose tissue (iWAT) from male Kmo-defficient mice and their wild-type littermate controls on chow and high-fat diets.
Project description:The experiment was designed to identify transcriptomic changes induced by selective EP4 agonism during experimental colitis. The mice received 1mg/ kg of the selective EP4 agonist, EP4-D or sterile phosphate buffered saline as vehicle control orally once per day concomitant with 3% DSS in drinking water. The DSS treatment was stopped on day 5 and mice were provided access to autoclaved drinking water for 3 additional days along with agonist and vehicle treatments once daily as described above. At the end of the treatment period, colon tissues from the mice were harvested, RNA isolated, and sequenced to identify transcriptional changes.