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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse hair follicle stem cell

ABSTRACT: Mouse keratinocytes were isolated from K15-EGFP transgenic mice for FACS sorting. RNA samples from EGFP-high and alpha-6 integrin positive cells (hair follicle stem cells) and from EGFP negative and alpha-6 integrin positive cells were used for Microarray analysis.

ORGANISM(S): Mus musculus

SUBMITTER: George Cotsarelis

PROVIDER: E-GEOD-1096 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Capturing and profiling adult hair follicle stem cells.

Morris Rebecca J RJ Liu Yaping Y Marles Lee L Yang Zaixin Z Trempus Carol C Li Shulan S Lin Jamie S JS Sawicki Janet A JA Cotsarelis George G

Nature biotechnology 20040314 4

The hair follicle bulge possesses putative epithelial stem cells. Characterization of these cells has been hampered by the inability to target bulge cells genetically. Here, we use a Keratin1-15 (Krt1-15, also known as K15) promoter to target mouse bulge cells with an inducible Cre recombinase construct or with the gene encoding enhanced green fluorescent protein (EGFP), which allow for lineage analysis and for isolation of the cells. We show that bulge cells in adult mice generate all epithelia ...[more]

PMID: 15024388

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Hair follicle stem cell gene profile

Project description:Mouse keratinocytes were isolated from K15-EGFP transgenic mice for FACS sorting. RNA samples from EGFP-high and alpha-6 integrin positive cells (hair follicle stem cells) and from EGFP negative and alpha-6 integrin positive cells were used for Microarray analysis. Keywords = stem cells Keywords = hair follicle Keywords = epidermis Keywords = alopecia Keywords: ordered

2004-04-01 | GSE1096 | GEO

Cancer stem cell features shift during skin squamous cell carcinoma progression

Project description:Cancer stem cells (CSCs) play a key role in tumour growth and metastasis. Although an expansion of CSC population was previously described in advanced cutaneous squamous cell carcinoma (SCC), it remains unknown whether CSC regulatory mechanisms also change through tumour progression to promote malignancy. Here, we generate mouse models of skin SCCs to study alterations in CSC regulation over progression. We found that features of CSCs change at late stage of progression, correlating with a switch from epithelial to mesenchymal tumour shape. beta-catenin and EGFR signalling are down-regulated, whereas autocrine FGFR1 and PDGFR alpha pathways are up-regulated in CSCs of advanced tumours. Inhibition of FGFR and PDGFR signalling repress malignant SCCs growth and metastasis, respectively. An enhanced epithelial-to-mesenchymal transition (EMT) program and over-expression of PDGFR alpha and FGFR1 are observed in malignant human skin SCCs, suggesting that these pathways are also induced over human SCC progression. Therefore, uncover signalling pathways controlling CSCs at specific stage of progression may improve the selection of more accurate targeted therapeutic strategies according to tumour stage, blocking SCC relapse and metastasis. Mouse models of skin SCCs progression were first generated. For that, individual samples of spontaneous or DMBA-TPA induced skin SCCs generated in K14-HPV16 mice were orthotopically engrafted in immunodeficient mice and then serially transplanted, generating lineages in which the progression from WD-SCCs to PD-SCCs was observed. Then, tumour cells expressing CD34 and alpha 6-integrin, previously identified as cutaneous cancer stem cells, were isolated by flow cytometry-sorter from WD-SCCs and PD-SCCs of different tumour lineages. RNA extraction and hybridization on Affymetrix microarrays was carried out to compare whole gene profiling of these populations of CSCs.

2016-01-08 | E-GEOD-59439 | biostudies-arrayexpress

Stump project-- mRNA basal layer keratinocytes; Volar versus non-volar acral skin gene expression

Project description:We were interested in defining the gene signature of volar skin. Punch biopsies of skin were split into epidermis and dermis after dispase treatment. Epidermis was trypsinized and sorted for alpha 6 integrin positive basal layer keratinocytes We collected RNA from basal layer keratinocytes of soles and backs of feet and submitted for Affymetrix Exon arrays. 2 replicates of each site from distinct human donors were included; total of 4 samples analyzed

2013-07-03 | E-GEOD-39500 | biostudies-arrayexpress

High Runx1 levels promote a reversible more differentiated cell-state in hair follicle stem cells during quiescence

Project description:Quiescent hair follicle (HF) bulge stem cells (SCs) differentiate to early progenitor (EP) hair germ (HG) cells, which divide to produce transit-amplifying (TA) matrix cells. EPs can revert to SCs upon injury, but whether this de-differentiation occurs in normal HF homeostasis (hair cycle), and the mechanisms regulating both differentiation and de-differentiation are unclear. Here we use lineage tracing, gain of function, transcriptional profiling, and functional assays to examine the role of observed endogenous Runx1 level changes in the hair cycle. We find that forced Runx1 expression implements hair degeneration (catagen) and simultaneously promotes changes in the quiescent bulge SC transcriptome towards a cell-state resembling the EP HG fate. This cell-state transition is functionally reversible. We propose that SC differentiation and de-differentiation are likely to occur during normal HF degeneration and niche restructuring in response to changes in endogenous Runx1 levels associated with SC location with respect to the niche. Freshly isolated skin cells were FACS sorted based on K15-GFP+/a6-integrin+/CD34- as hair germ and K15-GFP+/a6-integrin+/CD34+ bulge cells. Duplicate samlpes from mice at PD20 that showed telogen morphology throughout skin were used for RNA prepartion and Affymetrix analysis. Wild and transgenic samples after 1-day of doxycycline treatement were compared.

2014-01-15 | E-GEOD-53077 | biostudies-arrayexpress

Transcription profiling of mouse C2C12 myoblast induced with tetracycline to enhance integrin alpha7 expression

Project description:Analysis of C2C12 myoblast induced with tetracycline to enhance integrin alpha7 expression. Integrin alpha7 is the major laminin binding integrin in muscle cells. Enhancing its expression has been demonstrated to alleviate pathology in a murine model of Duchenne muscular dystrophy. Results of this study provide insights into the effects of increasing integirn alpha7 expression on muscle cells and possible side effects associate with enhancing integrin alpha7 in muscle cells. Experiment Overall Design: Triplicate biological smaples for each condition(induced and non-induced) C2C12 myoblast were used.

2008-06-16 | E-GEOD-8312 | biostudies-arrayexpress

Expression data from 13 week human fetal scalp epidermis sorted for expression of alpha 6 integrin and CD133

Project description:CD133 is expressed by a subpopulation of human fetal hair follicle placode cells during early hair development. Its expression, which is gradually lost as the placode matures, correlates with early morphogenesis. We used microarrays to analyze differences in gene expression between alpha-6 integrin+CD133+ basal cells and other human fetal epidermal populations to provide candidates defining this unique hair follicle placode subpopulation. 13 week human fetal scalp epidermis was sorted into 3 populations based upon expression of alpha 6 integrin and CD133 and RNA analyzed for gene expression.

2014-07-01 | E-GEOD-58393 | biostudies-arrayexpress

RNA-Seq in hair follicle stem cell lineage

Project description:We performed RNA_Seq on purified hair follicle stem cells (HFSCs)and their direct progenty, transit amplifying cells (TACs) using temorally and spatially regulated Cre lines. Consequences of loss of Bmpr1a in either HFSC (K15-CrePGR;Bmpr1a fl/fl), or TACs (1. Shh-CreER:Bmpr1a fl/fl or 2. K15-CrePGR;Bmpr1a fl/fl derived)

2014-09-30 | E-GEOD-61505 | biostudies-arrayexpress

Alternate covalent form of platelet αIIbβ3

Project description:The αIIbβ3 integrin receptor coordinates platelet adhesion, activation and mechanosensing in thrombosis and haemostasis. Using differential cysteine alkylation and mass spectrometry, we have identified a disulfide bond in the αIIb subunit linking cysteines 490 and 545 that is missing in about one in three integrin molecules on the resting and activated human platelet surface. This alternate covalent form of αIIbβ3 is pre-determined as it is also produced by human megakaryoblasts and bovine hamster kidney cells transfected with recombinant human integrin. From co-immunoprecipitation experiments, the alternate form selectively partitions into focal adhesions on the activated platelet surface. Its function was evaluated in bovine hamster kidney cells expressing a mutant integrin with an ablated C490-C545 disulfide bond. The disulfide mutant integrin has extended residency time in focal adhesions due to reduced rate of clathrin-mediated integrin internalisation and recycling that is associated with enhanced affinity of the αIIb subunit for clathrin adaptor protein-2. The alternate covalent form also has different conformational dynamics measured by monoclonal antibody binding and molecular dynamic simulations, which is reflected in reduced fibrinogen binding and outside-in signalling. These findings indicate that the αIIbβ3 integrin receptor is produced in different covalent forms that have different functions. The C490, C545 cysteine pair is conserved across all 18 integrin α subunits and the disulfide bond in αV in a cancer cell line is similarly missing, suggesting that this alternate integrin form and function is also conserved.

2022-02-17 | PXD025907 | Pride

Depletion of beta1 integrins in triple negative breast cancer cells causes extensive alterations in gene expression profiles

Project description:Interactions with the extracellular matrix (ECM) through integrin adhesion receptors provide cancer cells with physical and chemical cues that act in concert with growth factors to support survival and proliferation. Preclinical studies testing beta1 integrin antagonists in (breast) cancer models have shown inhibition of tumor growth and sensitization to radio- or chemotherapy and these strategies are currently evaluated in clinical trials. Here, we show that disruption of beta1 integrin-mediated ECM adhesion attenuates breast tumor growth but dissemination to the lungs from such small tumors can be markedly enhanced. beta1 integrin downregulation induces compensatory upregulation of beta3 integrins, but increased beta3 expression does not lead to enhanced lung metastasis. Instead, beta1 integrin downregulation in human and mouse triple negative, E-cadherin positive breast cancer cells elicits a switch from collective invasion to individual cell migration in 3D ECM. This involves alterations in the TGFbeta-BMP signaling network shifting the balance between miR-200 and ZEB, which causes a block in E-cadherin transcription. The switch is fully reversible: restored beta1 expression reinstates E-cadherin expression and cell cohesion. Moreover, restoring the network at the level of TGFbetaR, ZEB/miR-200 balance, or E-cadherin, restores cohesion and prevents the induction of lung metastasis without affecting tumor growth. These findings reveal that integrin-mediated ECM-attachments regulate a signaling network in control of epithelial characteristics that suppress metastatic spread. This raises concerns with respect to the use of beta1 integrins as cancer drug targets

2014-02-11 | E-MTAB-2185 | biostudies-arrayexpress

Pax2 Dependent Expression in the Intermediate Mesoderm at E11.5

Project description:Heterozygous and homozygous Pax2 E11.5 embryos were collected and the intermediate mesoderm was dissected and dispersed into single cells. The Pax2 positive cells also expressed EGFP, which was knocked into the Pax2 locus. EGFP positive cells were sorted by FACS and RNA isolated. We compared RNA expression levels in EGFP positive cells from Pax2 null and Pax2 heterozygote embryos.

2014-12-13 | E-GEOD-64129 | biostudies-arrayexpress

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