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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse mammary glands tumours from PEA3-null and wild-type PEA3 animals

ABSTRACT: Study of differences in gene expression profiles in the mammary glands tumours of PEA3-null and wild-type PEA3 mice induced by the Her2/Neu transgene. Experiment Overall Design: this experiment include 3 samples and 22 replicates

ORGANISM(S): Mus musculus

SUBMITTER: OGIC Info Ontario Genomics Innovation Centre (OGIC)

PROVIDER: E-GEOD-3765 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene Expression Study of Mammary Glands Tumours of PEA3-null and Wild-type PEA3 Mice

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2005-12-09 | GSE3765 | GEO

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Target genes associated to siRNA mediated repression of Pea3 and Erm transcription factors in mammary cancerous cells

Project description:Pea3 and Erm are transcription factors belonging to the ETS family which are involved in tumorigenesis, particularly mammary oncogenesis. To elucidate the mechanism of Pea3/Erm induced tumorigenesis in mouse mammary cancerous MMT cells (ATCC CCL-51), we have down-regulated Pea3 or Erm expression in this cell line by using small interfering RNA and performed a microarray analysis using Applied Biosystems AB1700 technology to define target associated genes. Keywords: Comparative transcriptome hybridization, small interfering RNA, mouse mammary cancerous cells Mouse mammary cancerous MMT cells (ATCC CCL-51) have been transfected with a siRNA control, an erm-directed siRNA or an pea3-directed siRNA. MMT cells were also used to determine the modifications due to the transfection protocol. These conditions correspond to four sample. Two biological replicate were used for the MMT cells (control sample), three biological replicates were used for the siRNA control (control sample) and erm-directed siRNA (reference sample) MMT cells and four biological replicates were used for the pea3-directed siRNA (reference sample) MMT cells. Transcriptome profiles (28.218 validated mouse genes) were acquired using Applied Biosystems AB1700 technology, which make use of chemiluminescence-based detection chemistry.

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Adenoviral Vector Transduction of Dendritic Cells is Inhibited by Plasma from Cancer Patients

Project description:Background: Pooled AB serum is often used as a media supplement for cell culture but it has the potential to transmit infectious diseases. To avoid this risk, we used autologous plasma as a media supplement for manufacturing dendritic cells (DCs) for cancer immunotherapy. We noticed inconsistences in the DCs and investigated its nature and cause. Methods: adHER2/neu DCs for 21 patients were manufactured from autologous peripheral blood monocytes which were treated with G-CSF and IL-4 for 3 days, transduced with Ad5f35HER2ECTM and then treated with lipopolysaccharide and IFN- for one day. The cells were cultured in RPMI-1640 supplemented with either 10% heat inactivated autologous or AB plasma. Results: Twenty-eight adHER2/neu DCs were manufactured for 21 patients using autologous plasma and 68 were manufactured for 20 of those patients using AB plasma. The expression of HER2/neu was less for DCs manufactured with autologous plasma (70.3±33.3% vs 86.1±22.8%, p<0.01). Manufacturing adHER2/neu DCs using monocytes from 3 healthy subjects and plasma from one patient with low HER2/neu expression (18%) resulted in low HER2/neu expression by all three DCs (13%, 16% and 23%). Analysis of the levels of 1322 proteins in 8 plasma samples associated with low HER2/neu expression and in 12 associated with high HER2/neu expression revealed that the levels of 14 predicted HER2/neu transduction efficiency. Conclusion: The manufacture of adHER2/neu DC using autologous plasma as a media supplement resulted in inconsistent HER2/neu expression. It is likely that variability in the levels of multiple proteins in autologous plasma contributed to low HER2/neu expression.

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Comparative Analysis of MMTV-neu tumors, preneoplastic MMTV-neu mammary gland, and Wild-type controls

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2005-05-03 | GSE2528 | GEO

RNA-seq of the lacrimal gland epithelial tissue in control vs Pea3-depleted tissue in mouse embyro

Project description:The goal of the study is to find the transcriptional targets downstream of Pea3-mediated signaling in the lacrimal gland epithelium. In order to look for the potential targets, we performed laser capture microdissection of the lacrimal gland epithelial tissue at embryonic day E14.5 from control embryos as well as mutant embryos containing epithelium-specific deletion of Pea3. After tissue harvest, RNA was extracted. Conversion to cDNA as well as amplification was performed by Clontech SMART-seq v4 Ultra low input RNA kit, as well as cDNA library construction was performed using Nextera XT DNA library preparation kit by core facility at Columbia university prior to RNA sequencing. After preparation of cDNA library, each sample was sequenced using Illumina platform.

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