Project description:Zfp800 was identified by Gene Co-expression Network analysis to be an interesting candidate gene involved in endocrine specification and β-cell maturation. We found that Zfp800 null mice exhibited early post-natal lethality, and at E18.5 their pancreata exhibited a reduced number of pancreatic endocrine cells, alterations in exocrine cell morphology, and marked changes in genes involved in protein translation, hormone secretion, and developmental pathways in the pancreas.
Project description:The canonical NF-κB pathway is active in 70% of all pancreatic cancer cases and NF-κB Essential Modulator (NEMO) is essential for the activation of this pathway. In our study, we used KC mice, which express the oncogenic KRAS and develop precancerous lesions termed Pancreatic Intraepithelial Neoplasias (PanINs), and KNeC mice, which express the oncogenic KRAS and have NEMO deleted in their pancreatic cells. These mice were injected with cerulein to promote the development of pancreatitis (cerulein dosage= 50μg/kg). Cerulein was injected at 8 hourly intervals for 2 days in total. The first injection day was when mice reached their sixth week of age and the second injection day was 3 days after the first injection day. Both KC and KNeC mice developed PanINs. At the age of 10 months, pancreata of KC and KNeC mice were analyzed. Using laser capture microdissection, PanINs from both groups were excised and their transcriptome was analyzed though RNA-seq.
Project description:We have carried out transcriptional profile analysis in WT MICE and bitransgenic Pdx1-cre/Kras*A MICE baring Pancreatic Ductal Adenocarcinoma Mouse models faithfully simulating human cancer are valuable for genetic identification of potential drug-targets but, among them, the most advantageous for practical use in subsequent preclinical testing of candidate therapeutic regimes are those exhibiting rapid tumor development. Considering that a KRAS mutation (predominantly in codon 12, such as KRASG12D; KRAS*) occurs with high frequency (~90%) in cases of human pancreatic ductal adenocarcinoma (PDA)1, we sought to develop a mouse PDA model that would exhibit high tumor incidence and short latency by ectopic overexpression of Kras*. Five WT mice and 6 bitransgenic Pdx1-cre/Kras*A MICE baring Pancreatic Ductal Adenocarcinoma were used to identify key genes in the formation of panceatic malignacies
Project description:Constitutive Kras and NF-kappaB activation is identified as signature alterations in human pancreatic ductal adenocarcinoma (PDAC). However, the mechanisms of constitutive NF-kappaB activation in KrasG12D-induced PDAC are not yet understood. Here, we report that pancreas-targeted IKK2/beta inactivation inhibited NF-kappaB activation and completely suppressed PDAC development in KrasG12D and KrasG12D;Ink4a/Arf mutant mice, demonstrating a genetic link between IKK2/beta and KrasG12D in PDAC inception. Our findings reveal that KrasG12D-activated AP-1 induces IL-1alpha, which in turn activates NF-kappaB and its target genes IL-1alpha and p62, to initiate IL-1alpha/p62 feedforward loops for inducing and sustaining NF-kappaB activity. Furthermore, IL-1alpha overexpression correlates with Kras mutation, constitutive NF-kappaB activity, and poor survival in PDAC patients. Therefore, our findings establish a pathway linking duel feedforward loops of IL-1alpha/p62 through which IKK2/beta/NF-kappaB is activated by KrasG12D. To study Kras-induced inflammatory responses and to identify differentially expressed genes between the pancreatic tissues of Pdx1-Cre;KrasLSL-G12D and Pdx1-Cre;KrasLSL-G12D;IKK2/betaF/F mice, cDNA microarray analysis was performed.
Project description:LFQ analysis of matrisome fractions enriched from 12 murine PDA or PanIN samples (samples 1 - 12) and a normal pancreas pool (samples 13) pooled from 10 pancreata of C57-Bl6 animals. Three independent enrichments have been conducted for each sample with each having had four injections.
Project description:Pancreata were excised from offspring of time-mated Wistar rats age 10-11 weeks (Taconic, Lille Skensved, Denmark) at embryonic day 20 (E20), immediately after birth (P0) and two days after birth (P2). After decapitation, individual pancreata were quickly dissected and placed directly in ice cold TRIReagent (Sigma-Aldrich, St Louis, MO, USA). RNA from individual pancreata was kept separately in order to verify the array results in pools of RNA as well as in individual animals by real time qPCR. 0.5 M-5g of RNA from each of 3 male and 3 female littermates were pooled at each time-point (E20, P0 and P2). This was repeated in each of three biological independent replicate experiments giving a total of 9 pooled samples, three at each time points
Project description:Pooled KRC (LSL-KrasG12D; Rb1L/L; Pdx1-Cre: oncogenic Kras and deleted Rb1 in the pancreas) cells derived from 2 month old mice were compared to pooled KC (LSL-KrasG12D; Pdx1-Cre: oncogenic Kras in the pancreas) cells derived from 8 month old mice.
Project description:Pancreatic ductal adenocarcinoma (PDAC) is strikingly resistant to conventional approaches. In this study, we report that the histone deacetylase associated SIN3B protein is required for activated KRAS-induced senescence in vivo using a mouse model of pancreatic cancer. We used microarray data to determine if SIN3B regulates KRAS-induced expression of the inflammatory response. Total RNA from Sin3Bp+/-KRaspG12D and Sin3Bp-/-KRaspG12D pancreas (two pancreata for each genotype) or PDEC (one for each genotype) was extracted and hybridized on Affymtrix microarrays.
Project description:Oncogene-induced senescence in early mPanIN lesions depends on intact RelA function in vivo. We investigated the difference in gene expression between KrasG12D pancreata with and without deletion of RelA. Pancreata from 7 day old mice were prepared, RNA was isolated and Affymetrix microarray expression analysis was performed.