Unknown,Transcriptomics,Genomics,Proteomics

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Global gene expression profiling in mouse plasma cell tumor precursor and bystander cells revels potential intervention targets for plasma-cell neoplasia

ABSTRACT: We extracted nascent plasma cell tumor (PCT) cells from within inflammatory granulomas (OG) isolated from intraperitoneal pristane-injected BALB/c.iMyc Eμ mice at five different time points during tumor progression. We used laser capture micro-dissection to collect incipient PCT cells and analyzed their global gene expression on Affymetrix Mouse Genome 430A microarrays. Two independent studies were performed with different sets of mice RNA was extracted from both laser capture samples and whole tissue sections at 5 different times after tumor initiation: 7 days, 17 days, 33 days, 49/46 days and 104/105 days in two independent experiments, studies 1 and 2

ORGANISM(S): Mus musculus

SUBMITTER: Thomas Bair

PROVIDER: E-GEOD-34078 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The prevalence of obesity has significantly increased worldwide. Therefore, this study aimed to evaluate the influence of obesity in the proteomic profile of periodontal ligament (PDL) tissues of rat first maxillary molars submitted to orthodontic tooth movement. The study was approved by the Ethical Committee on Animal Experimentation of the Sao Paulo State University, School of Dentistry at Araraquara, Brazil (protocol number 16/2015) and followed the Animal Research: Reporting of In Vivo Experiments guidelines. Therefore, for this study, we used ten male adult Holtzman rats (weight average – 300 g, approximately 8 weeks old). Animals were randomly assigned to one of the following experimental groups: 1 – M group: animals received a standard diet and 75 days after that, the first maxillary molar was randomly selected to be submitted to OTM for 15 days (n=5), and 2 – OM group: animals received a high-fat diet to induce obesity, and 75 days after that the first maxillary molar was randomly selected to be subjected to OTM for 15 days (n=5). All animals were kept under controlled temperature (22-25 °C), receiving water ad libitum with a 12-hour light/dark cycle. To induce obesity, animals of the OM group received a high-fat diet. In contrast to the standard diet (Labina/Purina, Ribeirão Preto, Brazil) administered to the M group. The high-fat diet contained around 3.82 kcal/g and was composed of 27.5 % of standard rat chow plus 25 % of peanuts, 25 % of milk chocolate, and 12.5 % sweet biscuits and contained 20 g of protein, 48 g of carbohydrate, 20 g of total fat, and 4 g of fiber per 100 g of diet. This feed was daily replaced and had started 75 days before baseline (M initiation). The procedure to induce OTM was by a closed coil nickel-titanium spring (Sentalloy, GAC Central Slip, New York, EUA) connected the maxillary first molar and central incisors with a steel wire (CrNi, 0.20 mm, 55.01.208; Morelli, Sorocaba, Brazi). Fifteen days after OTM, animals were euthanized by anesthetic overdose.

Project description:To elucidate mechanisms of cancer progression, we generated inducible human neoplasia in 3-dimensionally intact epithelial tissue. Gene expression profiling of both epithelia and stroma at specific time points during tumor progression revealed sequential enrichment of genes mediating discrete biologic functions in each tissue compartment. A core cancer progression signature was distilled using the increased signaling specificity of downstream oncogene effectors and subjected to network modeling. Network topology predicted that tumor development depends upon specific ECM-interacting network hubs. Blockade of one such hub, the b1 integrin subunit, disrupted network gene expression and attenuated tumorigenesis in vivo. Thus, integrating network modeling and temporal gene expression analysis of inducible human neoplasia provides an approach to prioritize and characterize genes functioning in cancer progression. There are 3 experiments in this study: (1a) a time course of human epidermal tissue transformed by oncogenic ER:H-Ras and IkBaM comprising days 0, 5, 20, and 35 post Ras activation using 4OHT; (1b) a matched time course of adjacent mouse stromal tissue during tumor progression; (2) Arrays comparing 4OHT-induced, Raf-1:ER/IkBaM transformed epidermal tissue and -4OHT controls; and (3) Arrays comparing Ras:ER/IkBaM grafts co-treated with 4OHT and either IgG control antibody or an anti-b1 integrin blocking antibody, P5D2 for 30 days. All arrays were done in biologic duplicate. Epidermal tissue co-expressing ER:H-RasG12V (ER:Ras) and IkBaM was regenerated on female scid/scid mouse recipients. Grafts were allowed to heal for at least 3 weeks before Ras activation via daily i.p. injections of 730ug of 4OHT (in 110ul of a corn oil/ethanol mixture). Duplicate grafts were harvested after 0, 5, 20, and 35 days of 4OHT treatment. Laser capture microdissection was utilized to independently isolate epithelial cells of the basal most layers and adajacent stromal tissue for each time point. RNA was subjected to one round of T7-based linear amplification (Ambion Message Amp II enhanced kit) and hybridized to either HG-U133A 2.0 or MG-430A affymetrix oligonucleotide arrays. Arrays from this experiment are labeled Ras and Stroma, respectively. The second experment was performed on Raf-1:ER/IkBaM expressing grafts with and without 4OHT treament for 30 days. The third experiment was done comparing ER:Ras/IkBaM grafts concomitantly treated with 4OHT and 1.5mg/week of either IgG control antibody or a mouse monoclonal blocking antibody against b1 integrin for 30 days. For experiments 2 and 3, HG-U133A 2.0 GeneChips were used; 4OHT treatment and RNA isolation/amplification was performed as above.

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Global gene expression profiling in mouse plasma cell tumor precursor and bystander cells revels potential intervention targets for plasma-cell neoplasia

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