Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mouse epidermis to investigate the differential regulation of RNA in skin from 7 week old female deltaNB-cateninER transgenic mice following 0, 1, or 7 days B-catenin activation in the epidermis

ABSTRACT: Expression of deltaNB-cateninER allows the time and duration of B-catenin activation to be controlled precisely through the application of 4OHT. We have shown that B-catenin activation in adult mouse epidermis stimulates de novo hair follicles and the formation of a new bulge and melanocyte niche. We utilize Affymetrix Mouse Genome 430A 2.0 oligonucleotide arrays to investigate the differential regulation of RNA in skin from 7 week old female deltaNB-cateninER transgenic mice following 0, 1, or 7 days B-catenin activation in the epidermis. RNA from age and sex matched wild-type littermates was used as a control.

ORGANISM(S): Mus musculus

SUBMITTER: Fiona Watt

PROVIDER: E-GEOD-1579 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Beta-catenin and Hedgehog signal strength can specify number and location of hair follicles in adult epidermis without recruitment of bulge stem cells.

Silva-Vargas Violeta V Lo Celso Cristina C Giangreco Adam A Ofstad Tyler T Prowse David M DM Braun Kristin M KM Watt Fiona M FM

Developmental cell 20050701 1

Using K14deltaNbeta-cateninER transgenic mice, we show that short-term, low-level beta-catenin activation stimulates de novo hair follicle formation from sebaceous glands and interfollicular epidermis, while only sustained, high-level activation induces new follicles from preexisting follicles. The Hedgehog pathway is upregulated by beta-catenin activation, and inhibition of Hedgehog signaling converts the low beta-catenin phenotype to wild-type epidermis and the high phenotype to low. beta-cate ...[more]

PMID: 15992546

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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.

Dataset Information

Transcription profiling of mouse epidermis to investigate the differential regulation of RNA in skin from 7 week old female deltaNB-cateninER transgenic mice following 0, 1, or 7 days B-catenin activation in the epidermis

Publications

Beta-catenin and Hedgehog signal strength can specify number and location of hair follicles in adult epidermis without recruitment of bulge stem cells.

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