Tumor suppressor effects of GR in skin: implication of stem cells

皮肤GR的抑癌作用：干细胞的意义

• 批准号: 8071063
• 负责人: Irina Budunova
• 金额: $ 27.83万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-20 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8071063
• 关键词: Address; Adverse effects; Affect; Agonist; Animals; Binding; Cell Cycle; Cell Maintenance; Cells; DNA Binding; Data; Development; Epithelial; Gene Expression; Gene Expression Regulation; Gene Proteins; Genes; Genetic; Glucocorticoid Receptor; Glucocorticoids; Goals; Growth; Hair follicle structure; Harvest; Homeostasis; Hormones; In Vitro; Keratin; Label; Laboratories; Ligands; Maintenance; Mediating; Mus; Pharmacological Treatment; Physiological; Play; Population; Research; Research Personnel; Resistance; Role; Signal Transduction; Skin; Skin Cancer; Skin Carcinogenesis; Skin Neoplasms; Stem cells; Stimulus; Testing; Topical application; Transactivation; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Tumor Promoters; Tumor Suppressor Proteins; Tumor-Derived; Work; cancer therapy; cell growth; clinical application; glucocorticoid-induced orphan receptor; in vivo; inhibitor/antagonist; innovation; insight; keratinocyte; metallothionein III; novel; overexpression; programs; research study; skin cancer prevention; transcription factor; tumor; tumorigenesis

DESCRIPTION (provided by applicant): Glucocorticoid hormones are potent inhibitors of skin carcinogenesis in mice. The activity of glucocorticoids is mediated by the known transcription factor, the glucocorticoid receptor (GR). Two major mechanisms of the regulation of gene expression by GR are DNA-binding dependent transactivation, and DNA-binding independent transrepression through negative interaction with other transcription factors. We found that keratin5.GR (K5.GR) transgenic animals in which GR was overexpressed and spontaneously activated in basal keratinocytes and in hair follicles, are resistant to ras-induced skin carcinogenesis both in terms of tumor multiplicity and tumor size. It is well accepted that mouse skin tumors derive from epithelial stem cells (SC) located in the bulge region of the hair follicle. It is also known that tumor promoters activate proliferation of quiescent epidermal SC located in the bulge. Our experiments showed that the number of putative SC in the bulge of K5.GR transgenic mice was almost two-fold less than in littermate controls. We also found that the clonogenic activity of SC-enriched keratinocyties isolated by FACS from skin of K5.GR mice was strongly decreased in comparison to SC-enriched keratinocytes isolated from w.t. animals. We transcriptionally profiled SC-enriched keratinocyties and mouse skin tumors harvested from K5.GR and w.t. mice using gene arrays. Unexpectedly, the expression of 70% of genes whose expression was affected by GR in SC and 86% of genes whose expression was affected by GR in skin tumors was inhibited by GR. These novel results suggest that negative gene regulation (transrepression) plays a key role in GR effects on epidermal SC maintenance and in GR tumor suppressor effects in skin. We propose to prove the hypotheses that (i) alteration of SC status including SC growth potential and sensitivity of SC to tumor promoting stimuli are important mechanisms of the tumor suppressor effect of GR in skin, and that (ii) transrepression activity of GR is critical for its tumor suppressor role. Pertinent to these research goals the Specific Aims are: 1). Define the effect of GR on proliferation of SC induced by tumor promoting stimuli in vivo using genetic and pharmacological approaches; 2). Determine the effect of GR on SC growth potential and sensitivity to differentiation in vitro using genetic and pharmacological approaches; 3). Identify the role of GR-induced gene transrepression in SC maintenance and in tumor suppressor effect in skin using KS.GRdim transgenic mice. This work is highly innovative because the role of GR and glucocorticoid-mediated signaling in the maintenance of epidermal SC has never been previously addressed. We expect that proposed studies will provide new insights into the maintenance of SC by glucocorticoids which are important physiological and pharmacological regulators of epidermal homeostasis. These studies will also significantly enhance our understanding of the role of stem cells in skin cancer resistance.

描述（由申请人提供）：糖皮质激素是小鼠皮肤致癌的有效抑制剂。糖皮质激素的活性是由已知转录因子，糖皮质激素受体（GR）介导的。通过GR调节基因表达的两个主要机制是DNA结合依赖性的反式激活，以及通过与其他转录因子负相互作用通过负相互作用而独立的DNA结合独立反映。我们发现，在基底角质形成细胞和毛囊中，GR过表达并自发激活的GR的转基因动物具有对Ras诱导的皮肤致癌性的抗性。很容易接受的是，小鼠皮肤肿瘤衍生自位于毛囊隆起区域的上皮干细胞（SC）。众所周知，肿瘤启动子激活位于凸起的静态表皮SC的增殖。我们的实验表明，K5.GR转基因小鼠的凸起中假定SC的数量几乎比同窝窝对照少了两倍。我们还发现，与从W.T.分离的富含SC的角质形成细胞相比。动物。我们从K5.Gr和W.T.收获的富含SC的角质形细胞和小鼠皮肤肿瘤进行了转录。使用基因阵列的小鼠。出乎意料的是，70％的基因的表达在SC中受GR的表达影响，而86％的基因表达受到皮肤肿瘤中GR影响的基因的表达受到GR的抑制。这些新的结果表明，阴性基因调节（反抑制）在GR对表皮SC维持和皮肤中GR肿瘤抑制作用中的影响起着关键作用。我们建议证明（i）SC状态的改变，包括SC的生长潜力和SC对促进肿瘤刺激的敏感性是GR在皮肤中肿瘤抑制效应的重要机制，而（ii）GR的转化活性对其肿瘤抑制作用至关重要。与这些研究目标有关的具体目的是：1）。定义GR对使用遗传和药理方法在体内促进刺激诱导的SC增殖的影响； 2）。使用遗传和药理方法确定GR对体外分化的SC生长潜力和敏感性的影响； 3）。使用Ks.grdim转基因小鼠，确定GR诱导的基因转化在SC维持和肿瘤抑制效应中的作用。这项工作具有很高的创新性，因为GR和糖皮质激素介导的信号在维持表皮SC中的作用从未得到过解决。我们预计，拟议的研究将为糖皮质激素对SC的维持提供新的见解，糖皮质激素是表皮稳态的重要生理和药理调节剂。这些研究还将显着增强我们对干细胞在皮肤癌耐药性中的作用的理解。

项目成果

Important role of kallikrein 6 for the development of keratinocyte proliferative resistance to topical glucocorticoids.

激肽释放酶 6 在角质形成细胞对局部糖皮质激素的增殖抵抗中发挥重要作用。

• DOI: 10.18632/oncotarget.9926
• 发表时间: 2016
• 期刊: Oncotarget
• 作者: Kishibe,Mari;Baida,Gleb;Bhalla,Pankaj;Lavker,RobertM;Schlosser,Bethanee;Iinuma,Sin;Yoshida,Shigetaka;Dudley,JoelT;Budunova,Irina
• 通讯作者: Budunova,Irina

Androgen receptor drives cellular senescence.

• DOI: 10.1371/journal.pone.0031052
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Mirochnik Y;Veliceasa D;Williams L;Maxwell K;Yemelyanov A;Budunova I;Volpert OV
• 通讯作者: Volpert OV

Selective Activator of the Glucocorticoid Receptor Compound A Dissociates Therapeutic and Atrophogenic Effects of Glucocorticoid Receptor Signaling in Skin.

• DOI: 10.15430/jcp.2015.20.4.250
• 发表时间: 2015-12-01
• 期刊: Journal of cancer prevention
• 影响因子: 2.5
• 作者: Klopot, Anna;Baida, Gleb;Budunova, Irina
• 通讯作者: Budunova, Irina

REDD1 functions at the crossroads between the therapeutic and adverse effects of topical glucocorticoids.

• DOI: 10.15252/emmm.201404601
• 发表时间: 2015-01
• 期刊: EMBO molecular medicine
• 影响因子: 11.1
• 作者: Baida G;Bhalla P;Kirsanov K;Lesovaya E;Yakubovskaya M;Yuen K;Guo S;Lavker RM;Readhead B;Dudley JT;Budunova I
• 通讯作者: Budunova I