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Ablation of Lgr5-positive cells in intestinal polyps from ApcMin Lgr5DTR mice

ABSTRACT: Ablation of Lgr5-positive cells in intestinal polyps from ApcMin Lgr5DTR mice

PROVIDER: PRJEB44931 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR5919740.fastq.gz	Fastqsanger.gz
	ERR5919750.fastq.gz	Fastqsanger.gz
	ERR5919760.fastq.gz	Fastqsanger.gz
	ERR5919770.fastq.gz	Fastqsanger.gz
	ERR5919780.fastq.gz	Fastqsanger.gz

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Ablation of Lgr5-positive cells in intestinal polyps from ApcMin;Lgr5DTR mice

Project description:We performed RNAseq on intestinal polyps from diptheria toxin-treated ApcMin;Lgr5DTR mice to investigate the effect of an acute selective pressure on stem cell populations in intestinal lesions. Lgr5+ cells in the ApcMin;Lgr5DTR mice were ablated with a single intraperitoneal dose of diphtheria toxin in saline (50 μg/kg), and samples were collected after 24 hours and after 5 days. Untreated ApcMin mice were used as control. Intestinal polyps were excised and collected for RNA sequencing.

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Project description:The generation of the Lgr5_EGFP_ires_CreERT2 knock-in mouse allows marking of Lgr5 positive cells of different tissues. Here we use these mice to sort Lgr5 positive cells and their daughter cells form intestinal adenomas and describe the expression profile of these two cell populations. Intestinal adenomas were generated by inducing Apc deletion through tamoxifen injection in Apc_fl/fl-Lgr5_EGFP_ires_CreERT2 mice. In these mice, GFP is expressed under the control of the Lgr5 promoter, leading to highest GFP levels in Lgr5 positive cells (GFP-high). Dividing GFP-high cells pass on Gfp two their daughter cells, thereby diluting the GFP. Daughter cells can therefore be isolated based on their lower GFP positivity (GFP-low). We sorted these two cell fractions (GFP-high and GFP-low) and compared them to each other on 4X44K Agilent Whole Mouse Genome dual colour Microarrays (G4122F). Three independent sorts were performed and RNA of GFP-high cells hybridized directly against RNA of GFP-low cells in three dye swap experiments, resulting in six individual arrays.

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Effect of MEK inhibiton on KrasQ61R mutant Lgr5-positive intestinal stem cells

Project description:We have previously shown that KRAS hyperactivation depletes the Lgr5-positive intestinal stem cells. Here, we aim to address whether reducing the MAPK pathway output, by using a MEK inhibitor (Cobimetinib) is sufficient to restore expression of Lgr5-positive intestinal stem cell markers.

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Project description:Background and aims: The transcription factor Stat3 has been considered to promote progression and metastasis of intestinal cancers. Methods: We investigated the role of Stat3 in intestinal tumors using mice with conditional ablation of Stat3 in intestinal epithelial cells (Stat3deltaIEC). Results: In the APCmin mouse model of intestinal cancer, genetic ablation of Stat3 reduced the multiplicity of early adenomas. However, loss of Stat3 promoted tumor progression at later stages leading to formation of invasive carcinomas which significantly shortened the lifespan of Stat3deltaIEC APCmin/+ mice. Interestingly, loss of Stat3 in tumors of APCmin/+ mice had no significant impact on cell survival and angiogenesis but promoted cell proliferation. A genome-wide expression analysis of Stat3-deficient tumors suggested that Stat3 negatively regulates intestinal cancer progression via the cell adhesion molecule Ceacam1. Conclusions: Our data suggest that Stat3 impairs progression of intestinal tumors. Therefore, detrimental effects on tumor progression have to be considered upon therapeutic targeting of the Stat3 signaling pathway in intestinal cancer.

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