Expression data from Wild-type (WT) and Alb/AEG-1 mice which are transgenic mice with hepatocyte-specific overexpression of AEG-1 (Astrocyte elevated gene 1)
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ABSTRACT: AEG-1 is overexpressed in human hepatocellular carcinoma (HCC) and positively regulates development and progression of HCC A transgenic mouse with hepatocyte-specific expression of human AEG-1 was generated using mouse albumin promoter/enhancer in B6/CBA background. Hepatocytes were isolated from WT and Alb/AEG-1 mice for RNA extraction and Affymetrix microarray hybridization.
Project description:Suppressor of cytokine signaling 3 (SOCS3) down-regulates several signaling pathways in multiple cell types, and previous data suggest that SOCS3 may shut off cytokine activation at the early stages of liver regeneration. We developed hepatocyte-specific Socs3 knockout (Socs3 h-KO) mice to directly study the role of SOCS3 during liver regeneration after 2/3 partial hepatectomy (PH). Socs3 h-KO mice demonstrate marked enhancement of DNA replication and liver weight restoration after 2/3 PH in comparison with littermate controls. Without SOCS3, signal transducer and activator of transcription 3 (STAT3) phosphorylation is prolonged, and activation of the mitogenic kinases extracellular signal-regulated kinase 1/2 (ERK1/2) is enhanced after PH. In vitro, we show that SOCS3 deficiency enhances hepatocyte proliferation in association with enhanced STAT3 and ERK activation after epidermal growth factor (EGF) or interleukin 6 (IL-6) stimulation. Microarray analyses show that SOCS3 modulates a distinct set of genes after PH, which fall into diverse physiologic categories. Using a model of chemical-induced carcinogenesis, we found that Socs3 h-KO mice develop hepatocellular carcinoma (HCC) at an accelerated rate. By acting on cytokines and multiple proliferative pathways, SOCS3 modulates both physiologic and neoplastic proliferative processes in the liver, and may act as a tumor suppressor. Experiment Overall Design: Hepatocyte-specific excision of the Socs3 gene was achieved by breeding Socs3 fl/fl mice with mice expressing the Cre recombinase transgene under control of the albumin promoter (Alb-Cre+), yielding Socs3 h-KO mice. Socs3 fl/fl, Alb-Cre- littermates were used as controls for all experiments, and are henceforth referred to as littermates. All mice (C57BL/6) were free of Helicobacter species, housed in a specific pathogen free facility with 12-h light/dark cycles with free access to standard food and water. 2/3 PH and sham operations were performed as previously described (15, 50) (n=3-6 mice per genotype per time point). Liver remnants were weighed after removal of necrotic stumps and sutures, and compared to post-operative body weight. For HCC experiments, a single i.p. injection of DEN (5mg/kg, Sigma) was performed 12-14 d after birth. For short time points, a single injection of DEN (100mg/kg) (31) was given to 4 wk old mice. At indicated time points, mice were sacrificed by CO2 inhalation. All animal studies were carried out under approved IACUC protocols at the University of Washington.
Project description:AEG-1 is overexpressed in human hepatocellular carcinoma (HCC) and positively regulates development and progression of HCC A transgenic mouse with hepatocyte-specific expression of human AEG-1 was generated using mouse albumin promoter/enhancer in B6/CBA background.
Project description:aCGH analysis of murine transgenic liver tissues affected with HCC, hybridized with age (12 months) and sex matched alb cre mice. Keywords: Array comparative genomic hybridization analysis (aCGH).
Project description:Double transgenic mice with hepatocyte-specific expression of AEG-1 and c-Myc show aggressive HCC compared to single transgenics. Gene expression was analyzed to understand the molecular mechanism by which AEG-1 and c-Myc cooperate to promote hepatocarcinogenesis. Livers were collected from naïve adult mice (3 mice/group). Total RNA was extrancted and subjected to RNA-Seq.
Project description:Alzheimer’s disease (AD) is the leading cause of late onset dementia. However, to date, no efficient therapy for AD is available. Here, we identified a neuroprotective role of the liver contributing to brain amyloid-β (Aβ) homeostasis. The hippocampus is a crucial region for learning and memory and is one of the brain areas most affected by AD. To investigate brain mechanisms underlying the neuroprotective role of the liver in AD pathology, we performed proteomics analysis of hippocampus extracted from 5×FAD::Alb-CreERT2;Ephx2−/− mice and control littermates. After quality control, we identified 67 protein expressions that were differently regulated between the two genotypes, including nine proteins in the Kyoto Encyclopedia of Genes and Genomes pathway of AD
Project description:aCGH analysis of murine transgenic liver tissues affected with HCC, hybridized with age (12 months) and sex matched alb cre mice. Keywords: Array comparative genomic hybridization analysis (aCGH). Independent HCC of MCL1-/- mice were hybridized with pooled wt mice. Mclâ1flox/flox mice (C57BL/6 background) were obtained from the Dana Farber Cancer Institute, Boston, USA (Opferman JT et al., Nature 2003) and bred to heterozygous Albumin-Cre mice (C57BL/6 background) which led to hepatocyte-specific deletion of Mcl-1. The mice develop severe chronic liver damage (Vick B et al., Hepatology, 2009).
Project description:Astrocyte elevated gene-1 (AEG-1) as a positive inducer of hepatocellular carcinoma (HCC). Transgenic mice with hepatocyte-specific expression of AEG-1 were challenged with N-nitrosodiethylamine (DEN) and developed multinodular HCC with steatotic features. Thus, we have identified the follwoing AEG-1 functions: induction of steatosis, inhibition of senescence and activation of coagulation pathway to augment an aggressive hepatocarcinogenic phenotype. Transgenic Mice liver tumors compared against WT mice liver tumors.
Project description:We have generated an inside-out genetic mouse model of liver cancer, the Alb-R26Met mice, in which slightly increased levels of the wild-type form of the MET Receptor Tyrosine Kinase (RTK) occurs specifically in the liver following the deletion of a stop cassette by the Cre recombinase. This genetic combination is obtained by crossing the R26stopMet with Alb-Cre mice. The Alb-R26Met mice spontaneously develop liver tumours and model the whole tumorigenic program. These tumours correspond to HCC patients belonging to the proliferative-progenitor subset. Importantly, the Alb-R26Met model recapitulates several features of HCC patients, including the molecular heterogeneity, the temporal heterogeneity of tumour onset, and the resistance to RTK inhibitors used in the clinic for HCC treatment. This unique genetic setting offered us the possibility to compare the transcriptomic profile of Alb-R26Met non-tumoral livers with Alb-R26Met advanced liver tumours. Outcomes offer an overall vision of how gene expression switches during a tumorigenic process modelled by the Alb-R26Met mice. Importantly, as tumorigenesis in the Alb-R26Met genetic setting occurs in an immune-competent context, outcomes offers as well the possibility to explore the profound changes in the immune microenvironment in the liver tumours, by performing deconvolution of RNAseq data.