Transcriptomic effects of fluocinolone acetonide treatment on mouse skin
Ontology highlight
ABSTRACT:
INSTRUMENT(S): NextSeq 500
ORGANISM(S): Mus musculus
SUBMITTER: Masaki Takasugi
PROVIDER: E-MTAB-11992 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Even though cutaneous atrophy is the major adverse effect of topical glucocorticoids, its molecular mechanisms are poorly understood. We found that glucocorticoids strongly increased the expression of REDD1 (regulated in development and DNA damage response 1), a stress-inducible inhibitor of mTOR, in mouse and human epidermis. We found that REDD1 knockout animals are partially resistant to glucocorticoid-induced epidermal and subcutaneous adipose atrophy which correlated with the protection of CD34+ follicular epithelial stem cells as well as p63+ keratinocyte progenitors in REDD1 knockout epidermis during chronic steroid treatment. At the same time, REDD1 knockout did not affect anti- inflammatory effect of glucocorticoids, as evaluated by ear edema test. Expression profiling revealed that ~ 50% of the glucocorticoid receptor (GR) target genes were not activated in epidermis of REDD1 knockouts, however, the negative effect of glucocorticoids on gene expression was similar to that in w.t. animals. Overall, our findings reveal a novel pathway in GR activation by its target gene/protein REDD1; and indicate an important role of REDD1 in glucocorticoid-induced skin atrophy, and maintenance of the epidermis and subcutaneous adipose. In addition, our findings form the foundation for the development of safer topical glucocorticoid treatment regimens by combining this therapy with REDD1 inhibitors. Seven wk old B6D2 females in the telogen stage of the hair cycle were shaved, and treated 3 days later. Glucocorticoid FA was applied topically (2 ug/animal) in 200 ul acetone to the back skin once or up to four applications every third day as described previously (Chebotaev et al., 2007b). Control animals were treated with acetone only. Skin was harvested 24 h after last FA application or 4-8 h after single FA application as indicated in Figure legends. To evaluate the anti-inflammatory effect of glucocorticoid FA in REDD1 KO and w.t. B6x129 mice, we used ear edema test (Schoepe et al., 2010; Schäcke et al., 2004; Park et al., 2006). Seven wk old female animals were pretreated with FA or vehicle applied to the back of the ear lobe one h before application of nonspecific contact irritant croton oil . Mice were sacrificed and ears were harvested nine hours after CO application, the time point at which we observed maximum ear edema in F1 C57Bl x 129 animals (data not shown). Four mm ear punch biopsies were immediately weighted to assess ear edema. Animals were injected i.p. with bromodeoxyuridine 1 hr before skin was harvested. Total RNA from murine epidermis and whole human skin was isolated with RiboPure kit (Ambion). Total RNA from murine s.c. adipose was isolated with RNeasy Lipid Tissue Kit (Qiagen). The RNA samples were treated with TURBOTM DNase (Ambion). Purity of RNA samples isolated from epidermis and s.c. fat was confirmed by the expression of adipose and keratinocyte cell markers.
2014-08-01 | E-GEOD-59151 | biostudies-arrayexpress
Project description:Female FVB/N and Tg.Ac (hemizygotes) mice 3-5 weeks of age were purchased from Harlan UK and Taconic UK, respectively. The mice were contained in an isolator and maintained at 19C with 50% relative humidity and a 12-hour light-dark cycle. Following a 1 week acclimatisation period the dorsal skin of the mice was shaved (approximately 8cm2 area) 2 days prior to commencing treatment. FVB/N mice (5 per group) were treated with dimethylbenz[a]anthracene (DMBA) (10ug/200ul in acetone) or vehicle control one week before promotion with TPA. TPA (2.5ug/200ul in acetone) was applied to the back of both FVB/N and Tg.Ac mice (5 per group) twice a week for up to 14 weeks; control mice received acetone. Mice were shaved and examined weekly for the presence of papillomas. At each time point (FVB/N mice 2, 4, 6 and 14 weeks of TPA promotion and Tg.Ac mice 4 days, 1, 2 and 9 weeks of TPA promotion) the mice were killed by cervical dislocation and the dorsal skin removed, a representative portion was fixed in 10% neutral buffered formalin and retained for histological analysis; the remainder was snap frozen in liquid nitrogen.
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Project description:Serpinb3a knockout mice show attenuated Transepidermal Water Loss (TEWL) and S100A8 expression (an early inflammatory marker) following treatment with Aspergillus fumigatus extract. The aim of the RNAseq experiments was to provided mechanistic insight for the role of Serpinb3a by identifying early differences in gene expression following allergen exposure. Wild type Balb/c and Serpinb3a null mouse backs were shaved and 24 hours later a patch containing 200ug of Aspergillus fumigatus extract was applied to the skin. After 3 days the patch was removed and the skin was harvested after an additional 24 hours.
2014-10-01 | E-GEOD-54640 | biostudies-arrayexpress
Project description:Cutaneous squamous tumors rely on autocrine/paracrine loops for proper fitness. Targeting this Achilles’ heel is therefore considered a potential avenue for patient treatment. However, the mechanisms that engage and sustain such programs during tumor ontogeny are poorly understood. Here, we show that two Rho/Rac activators, the exchange factors Vav2 and Vav3, control the expression of an epithelial autocrine/paracrine program that regulates keratinocyte survival and proliferation as well as the creation of an inflammatory microenvironment. Vav proteins are also critically involved in some of the subsequent autocrine signaling loops activated in keratinocytes. The genetic inactivation of both Vav proteins reduces tumor multiplicity without hampering skin homeostasis, thus suggesting that pan-specific Vav therapies may be useful in skin tumor prevention and treatment. The dorsal skin of WT and DKO mice (Vav2-/-;Vav3-/-) were treated with either one or four applications of phorbol ester 12-O-tetradecanoylphorbol-13 acetate (TPA) (6.8 nmol in 200 μl acetone) two days after shaving. As control, we applied 200 μl of acetone. Animals were euthanized 24 hours after treatment.
2013-07-30 | E-GEOD-40849 | biostudies-arrayexpress
Project description:In order to identify critical changes in the genetic network of Rage signaling induced by TPA throughout a 48 h kinetic, global gene expression analysis was performed. Total RNA from untreated, 24h acetone- and 6, 12, 24, 48 h TPA-treated wt and Rage KO back skin was prepared (n=3 independent animal experiments) and hybridized on 4x44K whole mouse genome oligonucleotide microarrays.
2010-11-29 | E-MTAB-205 | biostudies-arrayexpress
Project description:This experiment focused on the miRNA expressions within the tissues specific niches of mice afflicted with melanoma, compared to littermates without malignancy. BRafCA/ PtenloxP/ Tyr::CreERT2 or genetically matched littermates without Cre recombinase were treated with 2µl 4-hydroxytamoxifen (5mM), every other day, for on days 0, 2, and 4, on the shaved right flank of the abdomen of the animal. Mice slowly grow melanoma at the treated site, and some metastases distal to treatment location (eg. Ear or opposite side of abdomen). Endpoint criteria was designated as a Samples were taken when tumors reached 1.5mm diameter. Total RNA was isolated from tumors or tamoxifen treated healthy skin, skin adjacent to the treatment site, or the ear ( representing a metastatic site). Total RNA was prepared with FLAG-TAG kit and miRNA microarray 4.0 was performed.
2019-01-31 | E-MTAB-6373 | biostudies-arrayexpress
Project description:Gene expression in self-renewing epithelial tissues is controlled by cis- and trans-activating regulatory factors that mediate responses to exogenous agents capable of causing tissue damage, infection, inflammation, or tumorigenesis. We used network construction methods to analyze the genetic architecture of gene expression in normal mouse skin in a cross between tumor-susceptible Mus musculus and tumor-resistant Mus spretus. We demonstrate that gene expression motifs representing different constituent cell types within the skin such as hair follicle cells, haematopoietic cells, and melanocytes are under separate genetic control. Motifs associated with inflammation, epidermal barrier function and proliferation are differentially regulated in mice susceptible or resistant to tumor development. The intestinal stem cell marker Lgr5 is identified as a candidate master regulator of hair follicle gene expression, and the Vitamin D receptor (Vdr) links epidermal barrier function, inflammation, and tumor susceptibility. Keywords: Expression Quantitative Trait Loci A backcross was generated using male Mus spretus and female FVB/N mice; female F1 hybrids were mated with male FVB/N mice. Seventy-one backcross mice (8-12 weeks old) received a single dose of DMBA (25 µg per mouse in 200 µl acetone). Starting one week after the initiation tumors were promoted with TPA (200 µl of 10-4 M solution in acetone) twice weekly for 20 weeks. Initiation and promotion were performed on doral back skin. Normal tail skin was snap frozen when the animals were sacrificed. Tail epidermis from completely untreated Spretus, FVB, and Spretus x FVB F1 mice was also analyzed.
2008-12-31 | E-GEOD-12248 | biostudies-arrayexpress
Project description:Background: Germline polymorphisms can influence gene expression networks in normal mammalian tissues and can affect disease susceptibility. We and others have shown that analysis of this genetic architecture can identify single genes and whole pathways that influence complex traits including inflammation and cancer susceptibility. Whether germline variants affect gene expression in tumors which have undergone somatic alterations, and the extent to which these variants influence tumor progression, is unknown. Results: Using an integrated linkage and genomic analysis of a mouse model of skin cancer that produces both benign tumors and malignant carcinomas, we document major changes in germline control of gene expression during skin tumor development resulting from cell selection, somatic genetic events, and changes in the tumor microenvironment. The number of significant expression Quantitative Trait Loci (eQTLs) is progressively reduced in benign and malignant skin tumors when compared to normal skin. However, novel tumor-specific eQTLs are detected for several genes associated with tumor susceptibility, including Interleukin 18, Granzyme E, Sprouty homolog 2, and MAP kinase kinase 4. Conclusions: We conclude that the genetic architecture is substantially altered in tumors, and that eQTL analysis of tumors can identify host factors that influence the tumor microenvironment, MAP kinase signaling, and cancer susceptibility. A backcross was generated using male Mus spretus and female FVB/N mice; female F1 hybrids were mated with male FVB/N mice. Backcross mice (8-12 weeks old) received a single dose of DMBA (25 µg per mouse in 200 µl acetone). Starting one week after the initiation tumors were promoted with TPA (200 µl of 10-4 M solution in acetone) twice weekly for 20 weeks. Initiation and promotion were performed on doral back skin. DNA from 62 Carcinomas and matched untreated tails (used for normal DNA comparison) was obtained from tissue that was snap frozen when animals were sacrificed.
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