Project description:We report the application of bulk RNAseq assay in examining the dietary effect on mouse intestinal stem cell (ISC) subpopulation. Lgr5EGFPcreERT2 mice were fed with new western diet 1 (NWD1) or the control AIN76A diet for 3 or 12 months, to investigate differential dietary effect. To examine the reversibility of dietary effect, after 3 months of NWD1 feeding, the mice were switched to AIN feeding, and examined at 6 or 12 months old.
Project description:Bmi1tm1(cre/ERT2)Mrc/J (a.k.a. Bmi1creERT2 mice), B6.Cg-Gt(ROSA)26Sortm14(CAT-tdTomato)Hze/J (a.k.a. Rosa26tom mice) were crossed. The mice were fed with New Western diet 1 (NWD1) or Americal Instititue of Nutrition 76A (AIN76A or AIN) diet from weaning for 3 months. Tamoxinfen was injected to induce Bmi1 cell dedifferentiation, featuring the expression of tomato red fluoresence in the Bmi1 locus. Single intestinal epithelial cells were FACS isolated by the Epcam+, CD45- and Tomato (red fluorescence)+, and subject to scRNAseq assay.
Project description:We demonstrate that Bmi1-expressing cells represent the predominant epithelial ISC in early intestinal development and precede existence of the canonical Wnt-dependent Lgr5+ stem cell population. We reveal that early in development, Bmi1+ ISCs are highly proliferative, then transition to a slow-cycling state with the emergence of the Lgr5+ ISC. Combining single cell RNA-sequencing (scRNA-seq) with bulk population ATAC-seq (Assay for Transposase Accessible Chromatin sequencing) analyses, we identify that a non-canonical Wnt signaling pathway correlates with the temporal presence of the highly proliferative developmental Bmi1+ ISC population.
Project description:To investigate the intestinal ileum response to dietary tryptophan deficiency and the contribution of the intestinal microbiome in regulating these responses
Project description:Background: Human intestinal tissue samples are barely accessible to study potential health benefits of nutritional compounds. Dietary fibres have been descirbed to be beneficial for intestinal health. Therefore, in this study we explored the applicability of an in vitro model, namely human intestinal Caco-2 cells, to study the effect of dietary fibres on intestinal health. Transcriptomics was applied to obtain more insight into their mode of actions in the intestinal cells. Methods: Caco-2 cells were stimulated with 500 ug/ml dietary fibres and the maximal observed LPS contamination to serve as background control for 6 hours, total RNA was extracted and Affymterix Human Gene 1.1 ST arrays were used to analyze the gene expression profiles. To identify dietary fibre induced gene expression profiles in dietary fibre gene responses were compared to medium samples. Furthermore, to analyse differentiatlly affected pathways Ingenuite Pathway Analysis was employed. Results: Pathway analysis revealed a distinct separation between the dietary fibres. GOS and beta-glucan oat medium viscosity affected transcription of a lower amount of genes (gene cut-off p<0.05) and gen transcription changes suggest an increase in vesicle transport and altered cholesterol regulation. On the other hand, the other dietary fibres differentially regulated a larger numbers of genes (gene cut-off p<0.05) and all appeared related to immune responses. We observed an increase in intracellular and extracellular anti-bacterial pathways and production of cytokines specifically aimed at communication with the adaptive immune system. Conclusion: GOS and beta-glucan oat medium viscosity appeared to induce intestinal epithelial communication with the body, whereas the other dietary fibres appeared recognized as PAMP and induce epithelial cells to interact with the immune system.
Project description:Identification of BMI1, RYBP and H2AK119UB interactome in Glioblastoma (GBM) to elucidate BMI1 roles independent of the PRC1-complex in GBM.
