ABSTRACT: Cancer cachexia syndrome is observed in 80% of patients with advanced-stage cancer, and it is one of the most frequent causes of death. Severe wasting accounts for more than 80% in patients with advanced pancreatic cancer. Here we wanted to define, by using an microarray approach and the Pdx1-cre;LSL-KrasG12D;INK4a/arffl/fl, the pathways involved in muscle, liver and white adipose tissue wasting. The aim of our work was to characterize as extensively as possible the pathways activated by the pancreatic cancer-induced cachectic tissues. For this purpose, we generated and compared genome-wide expression profiles of white adipose tissue, skeletal muscle and liver, from Pdx1-cre;LSL-KrasG12D;INK4a/arffl/fl and LSL-KrasG12D;INK4a/arffl/fl mice at 10 weeks-old. Tissue samples by triplicate was obtained from liver, muscle and adipose tissues in both groups, controls and cachectic mice. Total RNA samples was processed and profiled on Affymetrix Mouse Gene 1.0 ST arrays as previously described (Cano et al, 2012)
Project description:We wished to investigate the role of E-cadherin loss in our mouse parietal cell/pre-parietal cell E-cadherin knock-out, p53 knock-out, oncogenic Kras induced model of gastric cancer. As such, we isolated RNA from stomach tissue from our E-cadherin knock-out model (Atp4b-Cre;Cdh1(fl/fl);Kras(LSL-G12D/+);Trp53(fl/fl);Rosa26(LSL-YFP/LSL-YFP)) and our E-cadherin heterozygous model (Atp4b-Cre;Cdh1(fl/+);Kras(LSL-G12D/+);Trp53(fl/fl);Rosa26(LSL-YFP/LSL-YFP)). We then performed a microarray on this stomach tissue from four independent mice of each genotype. Differentially expressed genes were identified and gene set overlap analysis was used to identify pathways enriched in one model over the other.
Project description:Although mutations in Kras are present in 21% of lung tumors, there is a high level of heterogeneity in phenotype and outcomes amongst lung cancer patients suggesting the importance of other pathways. Wnt/β-catenin signaling is a known oncogenic pathway that plays a well defined role in colon and skin cancer but its role in lung cancer remains unclear. We show that activation of Wnt/β-catenin in the bronchiolar epithelium of the adult lung does not promote tumor development by itself. However, activation of Wnt/β- catenin signaling leads to a dramatic increase in tumor formation both in overall tumor number and size compared to KrasG12D alone. We show that activation of Wnt/β- catenin signaling significantly alters the KrasG12D tumor phenotype resulting in a phenotypic switch from bronchiolar epithelium to the highly proliferative distal progenitors found in the embryonic lung. This is associated with a decrease in E- cadherin expression at the cell surface which may increase metastasis in Wnt/β-catenin signaling positive tumors. Together, these data suggest that activation of Wnt/β-catenin signaling in combination with other oncogenic pathways in lung epithelium may lead to a more aggressive phenotype due to the imposition of an embryonic distal progenitor phenotype accompanied by decreased E-cadherin expression. We performed microarray analysis of control murine lung, CC10-cre:KrasG12D, and CC10-cre:Ctnnb1ex3flox:LSL-KrasG12D double mutant micro-dissected murine lung tumors to determine their transcriptional phenotype. Lungs of five-month-old mice were PBS inflated and all the tumors in each lobe were dissected. The total number of tumors obtained from three out of the 5 pulmonar lobes of each animal was called a sample the other two lobes were saved in case there were problems and the array needed to be repeated. Trizol was used to isolate RNA for microarray analysis. Samples & Genotypes: control murine lung n=2 animals, CC10-cre:KrasG12D n=2 animals, and CC10-cre:Ctnnb1ex3flox:LSL-KrasG12D n=2 animals.
Project description:This study used Illumina RNA-sequencing to examine transcriptomic profile of mice with Klf5 knockout in context of oncogenic Kras expression. The study analyzed total RNA extracted from pancreas of mice 2 days after cerulein-induce pancreatitis. In addition, this study include 2 biological replicate of mice with each of the following genotypes: Ptf1aCreERTM, Ptf1aCreERTM;Klf5fl/fl, Ptf1aCerERTM;LSL-KrasG12D, and Ptf1aCreERTM;LSL-KrasG12D;Klf5fl/fl. Overall design: Transcriptomic profiling of pancreata from 2 Ptf1aCreERTM mice, 2 Ptf1aCreERTM;Klf5fl/fl mice, 2 Ptf1aCerERTM;LSL-KrasG12D mice, and 2 Ptf1aCreERTM;LSL-KrasG12D;Klf5fl/fl mice 2 days after cerulein-induced pancreatitis.
Project description:These experiments aimed to determine the global gene expression patterns in p120WT, p120HET, and p120NULL cells in the context on oncogenic mutant KrasG12D Overall design: p120ctn was deleted using in vitro lentiviral Cre recombinase in pancreatic ductal cells isolated from LSL-KrasG12D;Ctnnd1wt/wt (KC-p120ctnWT), LSL-KrasG12D;Ctnnd1fl/wt (KC-p120ctnHET) , and LSL-KrasG12D;Ctnnd1fl/fl (KC-p120ctnNULL) mice.
Project description:Nuclear Protein 1 (Nupr1) is a major actor of the cell stress response required for KrasG12D-driven formation of pancreatic intraepithelial neoplastic (PanINs) lesions in mice. We investigated the impact of Nupr1-depletion on the development and biology of murin pancreatic adenocarcinomas (PDAC) in the Pdx1-cre;LSL-KrasG12D;Ink4a/Arffl/fl (KIC) mice. We found that only one half of Nupr1-deficient mice developed PDAC. This is related to increased caspase 3 activity and low IER3 expression in Nupr1-deficient;KIC in the pancreas. Moreover, when Nupr1-deficient;KIC mice do develop PDAC, tumors present with impaired epithelial-to-mesenchymal transition (EMT). Transcriptoma analysis revealed that Nupr1-deficient and Nupr1wt;KIC PDACs presented enrichment of gene signatures of the human classical- and quasi-mesenchymal (QM)-PDAC respectively. Moreover, Nupr1-deficient;KIC PDACs shared with human classical-PDACs overexpression of Kras-activation genes. In addition, cells derived from Nupr1-deficient;KIC PDACs formed fewer microspheres in vitro compared to Nupr1wt;KIC cells, indicative of stemness impairment in the absence of Nupr1. Finally, we found that Nupr1-deficient;KIC cells were more sensitive to some anticancer drugs than their Nupr1wt counterpart. Hence, this study establishes the pivotal role of Nupr1 in PDAC progression after PanIN and in PDAC EMT in vivo, with an impact in PDAC cell stemness. As a consequence, according to absence or presence of Nupr1, KIC mice develop tumors that phenocopy human classical- or QM-PDAC, respectively, thus becoming attractive models for preclinical drug trials. We investigated the impact of the homozygous deletion of the Nupr1 gene on pancreatic adenocarcinoma development and biology in the Pdx1-cre;LSL-KrasG12D;Ink4a/Arffl/fl (KIC) mouse model.
Project description:To investigate the role of SHP2 (Ptpn11) in pancreatic carcinogenesis, murine pancreatic whole tissue RNA samples of 9 week old mice with the genotypes Ptf1a-Cre;LSL-KrasG12D (ID-labels Kxxx) and Ptf1a-Cre;LSL-KrasG12D;Ptpn11fl/fl (ID-labels Mxxxx) were analyzed by microarray.
Project description:Transcriptional profiling of mouse Prostate cancer cells comparing Pbsn-Cre LSL-KrasG12D P53L/L cells with Pbsn-Cre LSL-BrafV600E P53L/L cells, and to determine the effects of Kras or Braf mutantion on murine PCa gene expression. Overall design: Three-condition experiment, Pbsn-Cre LSL-KrasG12D P53L/L cells vs. Pbsn-Cre LSL-BrafV600E P53L/L cells vs. Pbsn-Cre P53L/L cells: 2 wild type replicates, 2 KrasG12D mutant replicates and 2 BrafV600E mutant replicates.
Project description:The Ink4a/Arf tumor supressors play crucial roles in inhibiting cell cycle progression at the G1/S checkpoint. Activating mutations in the KrasG12D oncogene is one of the most frequent changes in human cancer, with resultant constitutive mitogenic signaling within the cell. We generated cohorts of CD19Cre; KrasG12D/+; Ink4a/ArfL/+ mice to evaluate the combined contributions of Ink/Arf loss and KrasG12D activation in pre-B ALL lymphomas. In this data set we include the expression data obtained from pre-B ALL lymphomas isolated from mice with simultaneous deletion of the Ink/Arf locus and activation of KrasG12D oncogene (and CD19Cre; KrasG12D/+; Ink4a/ArfL/+ mice). Overall design: A total 10 mRNA samples were isolated from pre-B ALL arising in CD19Cre; KrasG12D/+; Ink4a/ArfL/+ mice.