ABSTRACT: Expression data of caerulein-treated wild-type mice and KrasG12D-mutated mice at different stages of pancreatic regeneration after inflammatory injury
Project description:Consecutive caerulein injections induce an acute pancreatitis in mice. Here, we recorded gene expression levels at different stages of pancreatic regeneration in wild-type mice as well as KrasG12D-mutated mice. Tissue was collected from mice pancreata, cell sorting was not performed. t=0h refers to the time where caerulein was injected. control referes to NaCl-treated samples (no caerulein). 13 time points were used for wild-type mice, 9 time points were used for KrasG12D-mutated mice. Multiple replicates were generated for each time point. We used Affymetrix GeneChip Mouse Gene 1.0 ST arrays.
Project description:Consecutive caerulein injections induce an acute pancreatitis in mice. Here, we recorded gene expression levels at different stages of pancreatic regeneration in wild-type mice as well as KrasG12D-mutated mice. Tissue was collected from mice pancreata, cell sorting was not performed. t=0h refers to the time where caerulein was injected. control referes to NaCl-treated samples (no caerulein).
Project description:Purpose: we used next generation sequencing to analyze gene expression profiles of pancreatic tissues from KrasG12D;Pdx1-Cre and miR-301a-/-;KrasG12D;Pdx1-Cre mice treated with caerulein. The goals of this study are to compare the different gene expression profiles of pancreatic tissue between KrasG12D;Pdx1-Cre and miR-301a-/-;KrasG12D;Pdx1-Cre mice treated with caerulein.
Project description:Transgenic KrasG12D mice can recapitulate pancreas intra-epithelial neoplasia (PanIN). Caerulein is a cholecystokinin analogue and induces acute pancreatitis when injected intra-abdominally. Caerulein-induced acute pancreatitis will accelerate PanIN progression in KrasG12D mice. We compared mRNA profile changes between KrasG12D mice with acute caerulein-induced pancreatitis and wild-type mice without acute pancreatitis. The experiment had two groups. Experiment group: KrasG12D mice with acute caerulein-induced pancreatitis (N=6). Three mice in experiment group received 1-week caerulein injection, and the other three mice received 2-week caerulein injection. All experiment group mice started to receive caerulein injection at 1-month of age, and were sacrificed at the last day of caerulein injection. Control group: wild-type mice without acute pancreatitis (N=6). The mice were sacrificed at 1.5-month of age. Whole pancreas tissue lysate samples were subjected to mRNA array assay.
Project description:Transgenic KrasG12D mice can recapitulate pancreas intra-epithelial neoplasia (PanIN). Caerulein is a cholecystokinin analogue and induces acute pancreatitis when injected intra-abdominally. Caerulein-induced acute pancreatitis will accelerate PanIN progression in KrasG12D mice. We compared mRNA profile changes between KrasG12D mice with acute caerulein-induced pancreatitis and wild-type mice without acute pancreatitis.
Project description:To study the role of Elp3 in pancreatic homeostasis and pancreatitis, pancreas tissues were collected from Elp3(fl/fl); Pdx-Cre and wild-type mice either untreated or after caerulein-induced pancreatitis. RNA sequencing was performed to identify transcriptional changes associated with Elp3 loss and pancreatic injury.
Project description:A subset of human pancreatic ductal adenocarcinoma cells (PDACs) is characterized by high Fosl1 expression and Fosl1 is linked to the control of pro-inflammatory pathways and growth of PDAC cells. To mimick the human disease in mice (> 90% of PDAC patients harbour Kras mutations) the mutated LSL-KrasG12D allele was combined with the pancreas specific Cre recombinase Ptf1aCre (p48Cre). The two pancreatic cancer cell lines (Ptf1aCre, LSL-KrasG12D/+) were isolated from these mice and used for transcriptomics studies. The two different murine pancreatic cancer cell lines (Ptf1aCre, LSL-KrasG12D) were treated with two different Fosl1 siRNAs and one control siRNA, each. 72h after transfection a sufficient knockdown was tested by immunoblotting and qPCR. Total mRNA was isolated and checked for integrity. According to manufacture's recommendation the samples were subjected to microarray analysis using the Affymetrix Mouse Gene ST 1.0 array chip to discover differentially expressed genes.
Project description:Purpose: Purpose: we used next generation sequencing to analyze gene expression profiles of pancreatic tissues from wild-type (WT) and miR-21-/- (miR-21 KO) mice treated with saline(control) or caerulein. The goals of this study are to compare the different gene expression profiles of pancreatic tissue between WT and miR-21 KO mice treated with caerulein. Methods: Female WT and miR-21 KO mice were administered 8-hourly intraperitoneal injection of 50μg/kg caerulein. Mice were sacrificed at 24 hours after the first caerulein injection and total RNA was extracted. mRNArna profiles were generated by deep sequencing WT treated with saline in single, WT treated with caerulein in duplicate, miR-21KO treated with caerulein in duplicate, using High-seq 2000 Illumina sequencing platform. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Results: After quality filtering of raw sequencing data, we obtained 32,797,327 (69.9%) out of 46,941,672 tags from WT mice treated with saline, 42,587,312 (87.1%) out of 48,919,213 tags from WT mice treated with caerulein, and 57,139,408 (85.1%) out of 67,111,415 tags from miR-21 KO mice treated with caerulein. We then mapped these tags to the mouse genome. We identified 1457 differential expressed genes (DEGs) between WT mice treated with caerulein and with saline, and 152 genes between WT mice and miR-21 KO mice treated with caerulein with a fold change more than 2.0 (up-regulation) or less than 0.5 (down-regulation). Altered expression of 16 genes was confirmed with qRT-PCR. Conclusions: Our study represents the first detailed analysis of pancreatic transcriptomes generated by RNA-seq technology. By using RNA-seq based transcriptome analysis, we identified 6 miR-21 target genes and 10 downregulated genes involved in pancreatic injury. Specifically, up-regulation of Pias3 and down-regulation of Hmgb1 when miR-21 was ablated coincided with reduced pancreatitis severity. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex pancreatic functions.
Project description:Acute pancreatitis (AP) is an inflammatory disease of pancreatic tissue caused by pancreatic acinar cell injury. However，its miRNA expression is not clear.A mouse model of acute pancreatitis was established by the establishment of Caerulein and LPS, high throughput miRNA sequencing was performed on the pancreatic tissues of mice in the normal group and model group. We found 25 miRNAs with relatively significant differential expression by analyzing the miRNA expression profile. Then we performed downstream target gene prediction for differentially expressed miRNAs. Analysising of KEGG, GO, GSEA and network interaction, it was found that its therapeutic effect was closely related to apoptosis, inflammatory response and autophagy process.
Project description:We are investigating the role of pancreatic pericytes in regulating acute pancreatitis using a mouse model consisting of WT and TG mice. The TG mice harbour a selective deletion of the Myd88 gene in pancreatic pericytes. Mice were treated with Caerulein to induce acute pancreatitis, and pancreatic pericytes were FACS-sorted one day after Caerulein treatment. RNA-seq analysis aims to determine differences in gene expression among the three groups: non-tg untreated, non-tg treated, and tg treated.
