Inflammatory bowel disease (IBD) is a debilitating chronic gastrointestinal condition resulting from an immune overreaction against microbiota. Intermittent inflammatory flares damage the gut and despite the cells’ efforts to repair the tissue and anti-inflammatory treatments, cellular programmes become altered over time. During IBD, fibroblasts – the main cells promoting wound healing – adopt a pro-inflammatory phenotype and secrete excessive extracellular matrix components, while epithelial cells trans-differentiate to acquire fibroblast-like properties. In addition, neurons modify their membrane potentials. Eventually, severe untreatable complications such as intestinal fibrosis, gut dysmotility and chronic pain appear. Therefore, understanding IBD pathomechanisms is a clinical unmet need.
To comprehend intestinal fibrosis, we isolate primary intestinal epithelial cells and fibroblasts from mice to induce IBD-relevant damage in vitro and characterize their phenotype (wound healing and contractibility assays, collagen secretion, IF, western blot, etc), and crosstalk (RNA-seq, confocal microscopy). Complementarily, we induce experimental colitis in genetically-modified mice lacking the expression of a DNA methyltransferase that seems crucial for fibrosis in epithelial cells or fibroblasts, or lacking the expression of key IBD autophagy and ER stress genes in neurons.
Inflammatory bowel disease (IBD) is a debilitating chronic gastrointestinal condition resulting from an immune overreaction against microbiota. Intermittent inflammatory flares damage the gut and despite the cells’ efforts to repair the tissue and anti-inflammatory treatments, cellular programmes become altered over time. During IBD, fibroblasts - the main cells promoting wound healing - adopt a pro-inflammatory phenotype and secrete excessive extracellular matrix components, while epithelial cells trans-differentiate to acquire fibroblast-like properties. In addition, neurons modify their membrane potentials. Eventually, severe untreatable complications such as intestinal fibrosis, gut dysmotility and chronic pain appear. Therefore, understanding IBD pathomechanisms is a clinical unmet need.
To comprehend intestinal fibrosis, we isolate primary intestinal epithelial cells and fibroblasts from mice to induce IBD-relevant damage in vitro and characterize their phenotype (wound healing and contractibility assays, collagen secretion, IF, western blot, etc), and crosstalk (RNA-seq, confocal microscopy). Complementarily, we induce experimental colitis in genetically-modified mice lacking the expression of a DNA methyltransferase that seems crucial for fibrosis in epithelial cells or fibroblasts, or lacking the expression of key IBD autophagy and ER stress genes in neurons.
General knowledge on aseptic techniques, cell culture work and in vitro assays.
Culture of enteric fibroblasts and epithelial cells.
Immunostaining, epifluorescence microscopy.
Assessment of intestinal tissue damage in tissue sections from mice.
Quantification of collagen using Sircol assay.
Experimental design and analyses.
Funded by the European Union. Views and opinions expressed are however those of the author(s) only and do not necessarily reflect those of the European Union or the European Education and Culture Executive Agency (EACEA). Neither the European Union nor EACEA can be held responsible for them.
