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Akt signalling leads to stem cell activation and promotes tumour development in epidermis.

ABSTRACT: A permantly active form of the oncogene Akt was expressed in the keratinocytes of the basal proliferative layer of the epidermis. Stem cells of the hair follicle expressing the cell surface marker CD34 were isolated. RNA form the CD34(+) and CD34(-) keratinocytes was extracted and and hybridized to Mouse Genome 430 2.0 Affymetrix arrays. Gene expression was compared between CD34(+) hair follicle stem cells and CD34(-) cells isolates from the back skin of K5-myrAkt transgenic mice.

ORGANISM(S): Mus musculus

SUBMITTER: Ramon Garcia-Escudero

PROVIDER: E-GEOD-48522 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Akt signaling leads to stem cell activation and promotes tumor development in epidermis.

Segrelles Carmen C García-Escudero Ramón R Garín Maria I MI Aranda Juan F JF Hernández Pilar P Ariza José M JM Santos Mirentxu M Paramio Jesús M JM Lorz Corina C

Stem cells (Dayton, Ohio) 20140701 7

Hair follicle stem cells (HF-SCs) alternate between periods of quiescence and proliferation, to finally differentiate into all the cell types that constitute the hair follicle. Also, they have been recently identified as cells of origin in skin cancer. HF-SCs localize in a precise region of the hair follicle, the bulge, and molecular markers for this population have been established. Thus, HF-SCs are good model to study the potential role of oncogenic activations on SC physiology. Expression of ...[more]

PMID: 24504902

Similar Datasets

Akt signalling leads to stem cell activation and promotes tumour development in epidermis.

Project description:A permantly active form of the oncogene Akt was expressed in the keratinocytes of the basal proliferative layer of the epidermis. Stem cells of the hair follicle expressing the cell surface marker CD34 were isolated. RNA form the CD34(+) and CD34(-) keratinocytes was extracted and and hybridized to Mouse Genome 430 2.0 Affymetrix arrays.

2014-01-26 | GSE48522 | GEO

Transcriptional analysis of apoptotic cerebellar granule neurons following rescue by Gip, Sp, Pacap and Igf1

Project description:Primary cultures of Cerebellar Granule Neurons (CGNs) have been extensively utilized to examine the signal transduction mechanisms underlying neuronal apoptosis. We conducted whole-genome expression profiling to decipher the transcriptional program controlling the apoptotic/survival switch in cerebellar granule neurons (CGNs) following the induction of apoptosis by serum and potassium deprivation and their rescue by gastric inhibitory polypeptide (Gip), substance p (Sp), insulin-like growth factor-1 (Igf1) or pituitary adenylyl cyclase-activating polypeptide (Pacap). Our results reveal the transcriptional changes intersecting neuronal apoptosis and survival and form the basis for further functional analyses and pharmacological exploitation to identify neuroprotective drugs. After six days âin vitroâ (DIV), extracellular KCl of CGNs was shifted from 25 to 5 mM for neuronal apoptotic death induction. After two washes with serum-free BME containing 5 mM KCl, neurons were incubated with the same medium for 6 h (K5), while control neurons were incubated with serum free medium supplemented with 25 mM KCl (K25). K5 neurons were also treated with a maximal effective dose of Gip, Sp, Igf1 and Pacap. Four biological replicates (derived from the same litter) for each of the experimental conditions (K25, K5, K5 + Gip; K25, K5, K5 + Sp; K25, K5, K5 + Igf1; K25, K5, K5 + Pacap) were analyzed.

2016-07-03 | E-GEOD-67788 | biostudies-arrayexpress

Transcription profiling of mouse epidermal cells sorted into three populations based on alpha 6 integrin, CD34 or Sca-1 surface marker expression

Project description:Epidermal keratinocytes were isolated from adult murine skin and FACS sorted into three populations based on their surface maker expression of alpha 6 integrin, CD34 and Sca-1. <br>- Alpha 6 positive cells constitute the entire population of basal or proliferative keratinocytes from the interfollicular epidermis and hair follicle. Both BIB and Bulge cells are contained within the total population of alpha 6 positive cells. <br>- BIB cells reside in the hair follicle in a region between the infundibulum and bulge and these cells are characterized by low alpha 6 integrin surface levels and are negative for CD34 and Sca-1 surface proteins. <br>- Bulge cells are derived from the bulge region of the hair follicle and are characterized by low + high alpha 6 integrin surface levels and are positive for CD34 but negative for Sca-1.

2009-11-16 | E-MEXP-928 | biostudies-arrayexpress

A Functional Role of RB-Dependent Pathway in the Control of Quiescence in Adult Epidermal Stem Cells Revealed by Genomic Profiling

Project description:Continuous cell renewal in mouse epidermis is at the expense of a pool of pluripotent cells that lie in a well defined niche in the hair follicle known as the bulge. To identify mechanisms controlling hair follicle stem cell homeostasis, we developed a strategy to isolate adult bulge stem cells in mice and to define their transcriptional profile. We observed that a large number of transcripts are underexpressed in hair follicle stem cells when compared to non-stem cells. Importantly, the majority of these downregulated genes are involved in cell cycle. Using bioinformatics tools, we identified the E2F transcription factor family as a potential element involved in the regulation of these transcripts. To determine their functional role, we used engineered mice lacking Rb gene in epidermis, which showed increased expression of most E2F family members and increased E2F transcriptional activity. Experiments designed to analyze epidermal stem cell functionality (i.e.: hair regrowth and wound healing) imply a role of the Rb-E2F axis in the control of stem cell quiescence in epidermis. Total RNA was prepared from CD34+ and CD34- keratinocytes obtained from pools of at least six mice per MACS isolation procedure. A total of three isolation procedures were performed. RNA extracted from each fraction (CD34+ / CD34-) was assessed for its quality using Agilent 2100 Bioanalyzer (Agilent Technologies) and hybridized to two Affymetrix Mouse 430 2.0 arrays, to ensure reproducibility of the process. This resulted in a total of 12 chips (6 CD34+ / 6 CD34-).

2010-03-20 | E-GEOD-19448 | biostudies-arrayexpress

RNA sequencing of isolated murine basal and suprabasal hair follicle bulge stem cells

Project description:In most tissues, stem cells (SCs) comprise a heterogeneous cell population. Previous reports showed that slow cycling label-retaining cells (LRCs) are present in two prominent hair follicle SC populations, basal and suprabasal bulge SCs (bBSCs, sbBSCs). CD34+/itga6low sbBSC and neighbouring CD34+/itga6high bBSCs seem similar with both SC populations able to self-renew in vitro and to generate new hair follicles when grafted. Until now, functional differences have not been reported and molecular mechanisms underlying a positional encoded, intrinsic bulge SC heterogeneity and the relations to SC plasticity and tissue function are not well understood. Following fluorescence activated cell sorting for CD34 and integrin alpha6 of keratinocytes from 7 weeks old mice, we provide sequencing data for two prominent hair follicle SC populations, bBSC and sbBSC, suggesting distinct and overlapping functions in skin homeostasis.

2021-07-01 | GSE166921 | GEO

Expression profile of Gli1-expressing keratinocytes from mouse skin

Project description:To assess if Hedgehog (Hh) responding cells in the skin have a unique expression profile, isolated keratinocytes that express the Hh response gene Gli1 were collected by FACS and their gene expression was compared to sorted CD34-expressing cells from the middle bulge region of the hair follicle and to cells from the interfollicular epidermis (IFE) by hybridization of isolated RNA to gene expression microarrays. Dissociated keratinocytes isolated form back skin of Gli1-eGFP/+ mice (2 mice pooled for each replicate) in adult telogen were sorted based on GFP expression and immunostaining for CD34. Only viable, single cells with immunostaining for α6 Integrin (to mark basal keratinocytes) were collected. GFP(+) cells were collected as the Gli1 cohort. GFP(-) CD34(+) cells were collected as the CD34 cohort, and GFP(-) CD34(-) cells were collected as interfollicular epidermis (IFE) cohort. Total RNA was extracted from each cell population and labeled for hybridization to gene expression microarrays. The experiment was preformed in triplicate, however RNA from the Gli1(+) cells in one replicate was of insufficient quality to analyze. For staining of isolated keratinocytes, cells were incubated on ice for 1 hour in SMEM containing 1%BSA and 15μL of anti-CD49f-PE antibody (BDBioscience) plus 5μL anti-CD34-APC antibody (eBioscience) for each 100μL total volume for each 2 million cells. Immediately prior to sorting, DAPI (Invitrogen) was added to the washed cells at 1μg/mL final concentration.

2011-12-01 | E-GEOD-28108 | biostudies-arrayexpress

Transcription profiling of mouse CD34-enriched keratinocyte stem cells

Project description:Recent developments in animal models (Morris et al., 2004; Tumbar et al., 2004) as well as the discovery of cell surface markers (Jones and Watt, 1993; Tani et al., 2000; Trempus et al., 2003; Nijhof et al., 2006) have made it possible to isolate living epidermal hair follicle stem cells (HFSCs) from mouse skin, facilitating the study of the biological and molecular features inherent to HFSCs. A complexity of stem and progenitor cell populations within the hair follicle has been revealed. Here, we report comprehensive profiling of mouse CD34-expressing HFSCs using the Agilent mouse oligo microarray platform in order to extend and enrich the existing HFSC databases. Experiment Overall Design: Total RNA was prepared from CD34(+) and CD34(-) keratinocytes obtained from three biological replicates, labeled with 2 different fluorescent dyes, and hybridized to the Agilent oligo-microarrays containing ~22,000 mouse genes and Expression Sequence Tag (EST) probes (supplemental methods). The experiment contained technical dye-flips of each pairwise comparison.

2008-06-15 | E-GEOD-7690 | biostudies-arrayexpress

Mouse bulge (CD34+CD200+CD49+) versus secondary hair germ (CD34-CD200+CD49+) versus interfollicular epidermis (CD34-CD200-CD49+)

Project description:Mouse back skin was disassociated to single cells, sorted by cell surface markers and tested by microarrray To compare the gene expression of mouse bulge (CD34+CD200+CD49+) versus secondary hair germ (CD34-CD200+CD49+) versus interfollicular epidermis (CD34-CD200-CD49+) xx Bald scalp retains hair follicle stem cells but lacks CD200-rich and CD34-positive hair follicle progenitor cells Androgenetic alopecia (AGA) or common baldness results from a marked decrease in hair follicle size. This miniaturization may be related to loss of hair follicle stem or progenitor cells. To test this hypothesis, we analyzed bald and non-bald scalp from the same individuals for the presence of hair follicle stem and progenitor cells using flow cytometry to quantitate cells expressing CYTOKERATIN 15 (KRT15), CD200, CD34 and ALPHA-6-INTEGRIN (ITGA6). High levels of KRT15 expression correlated with stem cell properties of small cell size and quiescence. Cells with the highest level of KRT15 expression were maintained in bald scalp; however, distinct populations of CD200high ITGA6high cells and CD34-positive cells were markedly diminished. Consistent with a progenitor cell phenotype, the diminished populations localized closely to the stem-cell rich bulge area but were larger and more proliferative than the bulge stem cells. In functional assays, analogous CD200 high /Itga6 high cells from murine hair follicles were multipotent and generated new hair follicles in skin reconstitution assays. These findings suggest that a defect in stem cell activation plays a role in the pathogenesis of AGA. 4 independent biologic replicates (each pooled from 3 distinct mice) were sorted for Mouse bulge (CD34+CD200+CD49+) versus secondary hair germ (CD34-CD200+CD49+) versus interfollicular epidermis (CD34-CD200-CD49+)

2011-01-12 | E-GEOD-21568 | biostudies-arrayexpress

Epidermal overexpression of ∆NLef1 leads to deregulation of epidermal stem cell fate

Project description:In adult K14ΔNLef1 mouse, the overexpression of ∆NLef1, a B-catenin dominat negative, in basal keratinocytes leads to the conversion of hair follicles into multilayered epithelial cysts and ectopic sebaceous gland. To uncover in vivo changes in gene expression associated to ΔNLef1 activity, we compared the expression profiles of basal keratinocytes (Itga6+) and bulge stem cells (cd34+/itga6+) in wild type and K14ΔNLef1 transgenic mice. Cells were collected from 9.5 week old mice, when the hair follicle are in the resting (telogen) phase of the hair growth cycle. At this stage the ΔNLef1 phenotype is not yet evident enabling us to detect early molecular events.

2019-03-08 | GSE118073 | GEO

Knockout of GPx4 gene in mouse keratinocyte

Project description:Comparative analysis of gene expression in cultured primary keratinocytes isolated from newborn control (K14-cre; GPx4fl/+) and knockout (K14-cre; GPx4fl/fl) mice. Selenoproteins are essential for skin function, as targeted abolition of selenoproteins in epidermal tissue results in newborn mice manifesting gross abnormalities of skin and hair, accompanied by retarded growth and premature death. To investigate whether lack of a single selenoprotein could induce similar phenotypic effect in mice, we generated keratinocyte-specific knockout mice lacking glutathione peroxidase 4 (GPx4), an essential selenoprotein in skin, to examine phenotypic changes resulting from the lack of GPx4 in skin. Ablation of GPx4 results in focal alopecia and disturbed hair follicle morphogenesis, with GPx4 being essential during early stages of hair follicle morphogenesis as well as for keratinocyte adhesion and proliferation in culture. We have generated mice with selective removal of the GPx4 gene in keratinocytes under the control of Keratin-14-cre (K14-cre) promoter. Comparative microarray analysis was performed on RNA samples taken from pooled primary keratinocytes from knockout and control mice from the same litter. Array replicates were performed using RNA samples from three different litters.

2012-12-06 | E-GEOD-34215 | biostudies-arrayexpress

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