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癌发生的有效抑制剂。糖皮质激素的活性由已知的转录因子糖皮质激素受体（GR）介导。 GR 调节基因表达的两个主要机制是 DNA 结合依赖性反式激活，以及通过与其他转录因子的负相互作用而实现的 DNA 结合独立反式抑制。我们发现，在基底角质形成细胞和毛囊中GR过度表达并自发激活的keratin5.GR（K5.GR）转基因动物，在肿瘤多样性和肿瘤大小方面都能够抵抗ras诱导的皮肤癌发生。人们普遍认为，小鼠皮肤肿瘤源自位于毛囊隆起区域的上皮干细胞（SC）。还已知肿瘤启动子激活位于凸起处的静止表皮SC的增殖。我们的实验表明，K5.GR 转基因小鼠凸起部分的推定 SC 数量几乎比同窝对照小鼠少两倍。我们还发现，与从野生型小鼠分离的富含 SC 的角质形成细胞相比，通过 FACS 从 K5.GR 小鼠皮肤分离的富含 SC 的角质形成细胞的克隆形成活性大大降低。动物。我们对从 K5.GR 和 w.t. 收获的富含 SC 的角质形成细胞和小鼠皮肤肿瘤进行了转录分析。使用基因阵列的小鼠。出乎意料的是，SC中70%的表达受GR影响的基因的表达和皮肤肿瘤中86%的表达受GR影响的基因的表达被GR抑制。这些新颖的结果表明，负基因调控（反式抑制）在 GR 对表皮 SC 维持的影响以及 GR 对皮肤肿瘤抑制的影响中发挥着关键作用。我们建议证明以下假设：(i) SC 状态的改变，包括 SC 生长潜力和 SC 对肿瘤促进刺激的敏感性是 GR 在皮肤中抑癌作用的重要机制，并且 (ii) GR 的反式抑制活性至关重要因其具有抑癌作用。与这些研究目标相关的具体目标是： 1)。使用遗传和药理学方法定义 GR 对体内促肿瘤刺激诱导的 SC 增殖的影响； 2）。使用遗传和药理学方法确定 GR 对 SC 生长潜力和体外分化敏感性的影响； 3）。使用 KS.GRdim 转基因小鼠鉴定 GR 诱导的基因反式抑制在 SC 维持和皮肤肿瘤抑制作用中的作用。这项工作具有高度创新性，因为 GR 和糖皮质激素介导的信号传导在维持表皮 SC 中的作用以前从未被解决过。我们期望所提出的研究将为糖皮质激素维持 SC 提供新的见解，糖皮质激素是表皮稳态的重要生理和药理学调节剂。这些研究还将显着增强我们对干细胞在皮肤癌抵抗中作用的理解。

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