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:We have studied pancreas samples in pancreas from a mouse model of acute pancreatitis, comparing wild-type animals and TRP14 KO animals. Samples are extracted with N-ethylmaleimide to block reduced cysteines, and no reducing agent is used, with a search including cysteinylated peptides. Acute pancreatitis was induced in 12 weeks-old mice (wild-type and KO for TRP14) by seven intraperitoneal injections of cerulein (50 µg/kg body weight) at 1 h intervals. Then, 1 h after the last injection, animals were euthanized under anesthesia with 3% isoflurane, exsanguinated, and the pancreas was immediately removed and processed.
Project description:To investigate the underlying changes during acinar-to-ductal metaplasia induced by different oncogenes and by acute pancreatitis, pancreatic tissue was isolated from 10 week old control wt, KrasG12D, Pi3kCAH1047R and Mek1dd mice and from mice of the same genotypes after induction of acute pancreatitis.
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:Autoimmune pancreatitis (AIP) is a disease with unclear immunologic triggers. This study shows that the pancreatic stellate cells are involved in the regulation of the immune response and can cause autoimmunity when the NF-κB signalling in these cells is disrupted.
Project description:Mouse pancreas from wild type and MistKO animals were induced either with caerulein or saline as control and processed for RNA. Targets from three biological replicates of each were generated and the expression profiles were determined using Affymetrix Mouse Expression chips 430. Comparisons between the sample groups allow the identification of genes with differential expression patterns of genes which might contribute to pancreatitis.
Project description:Autoimmune pancreatitis (AIP) is a disease with unclear immunologic triggers. This study shows that the pancreatic stellate cells(PSCs) are involved in the regulation of the immune response and can cause autoimmunity when the NF-κB signalling in these cells is disrupted. The PSCs were isolated from animals which show autoimmune pancreatitis (NEMO knockout group) or chronic pancreatitis (NEMO wildtype group).
Project description:Targeting transcription replication conflicts, a major source of endogenous DNA double stranded breaks and genomic instability, could have important therapeutic implications. When transcription and DNA replication machineries encounter each other on chromosomes, one way to resolve the conflict is temporarily dislodging RNA polymerase II from the conflict sites to enable the replication fork to proceed. Proliferating cell nuclear antigen (PCNA), an essential component of replisomes, plays a critical role in this process. We discovered a small molecule ligand (AOH1996) of PCNA, which targets a binding pocket adjacent to the outer surface region of PCNA known to interact with PIP box and APIM motif proteins. Orally administrable, AOH1996 suppresses tumor growth but causes no discernable side effects. In addition, we found that AOH1996 treatment can stabilize interaction between PCNA and RPB1, the largest subunit of RNA polymerase II. To determine whether the effect of AOH1996 is mediated through affecting PCNA interaction with RPB1, we mutated Y418 within the APIM motif of RPB1 to an alanine in the SK-N-AS cell line by CRISPR and compared changes in protein expression profiles caused by AOH1996 in the parent and mutant SK-N-AS cells.
Project description:MOLT4 cells in log phase (1x106 cells/ml) were irradiated with 4 Gy at room temperature, at a dose rate of 2.45 Gy/minute with a 137Cs ?-irradiator. After 4 hours, actinomycin D (5 ?M) was added to block transcription. Aliquots of culture were removed at hourly intervals and RNA was extracted (Trizol; Invitrogen). RNA and cRNA quantity and quality was determined by Nanodrop spectrophotometer and Bioanalyser 2100 (Agilent). Transcript levels were determined at different time points using Affymetrix Human U133A microarrays.<br><br>
Project description:Given that mice transplanted with glucocorticoid-resistant T cells display more severe disease symptoms, the idea was that an RNA-seq comparison between mice transplanted with either wildtype or glucocorticoid-resistant T cells could yield genes involved in disease progression and potential therapeutic targets.