Local Role for Steroids in Regenerative Growth in Drosophila

类固醇在果蝇再生生长中的局部作用

• 批准号: 10373939
• 负责人: Douglas Emmons Terry
• 金额: $ 4.68万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373939
• 关键词: Adult; Alleles; Anabolism; Androgens; Animals; Apoptotic; Back; Binding; Biological Models; Biology; Cells; Characteristics; Complex; Crustacea; Data; Development; Drosophila genus; Drosophila melanogaster; Ecdysone; Ecdysterone; Epithelial; Estrogens; Gene Expression; Gene Proteins; Genes; Genetic Transcription; Goals; Growth; Homologous Gene; Hormones; Human; Immunofluorescence Immunologic; Impairment; In Situ Hybridization; Injury; Laboratories; Ligand Binding Domain; Ligands; Light; Link; Malignant Neoplasms; Malignant neoplasm of prostate; Measures; Methods; Modeling; Molecular; NCOA3 gene; Natural regeneration; Nuclear Hormone Receptors; Oncoproteins; Orthologous Gene; Output; Pathologic; Pathway interactions; Pattern; Peripheral; Physiologic pulse; Porifera; Production; Proliferating; Proteins; Publishing; RNA; RNA Interference; Receptor Activation; Receptor Signaling; Regenerative pathway; Regenerative research; Reporter; Role; Signal Pathway; Signal Transduction; Site; Skin wound healing; Stanolone; Stem Cell Development; Stem Cell Factor; Steroid Receptors; Steroids; Surgical wound; System; Temperature; Testing; Therapeutic; Tissues; Transcription Coactivator; Transgenes; Tumor Suppressor Genes; Wing; Work; blastema; clinically significant; coactivator-associated arginine methyltransferase 1; cofactor; disc regeneration; ecdysone receptor; epithelial stem cell; epithelial wound; epithelium regeneration; experimental study; fly; genetic manipulation; germline stem cells; hormonal signals; insight; limb regeneration; malignant breast neoplasm; nano; notch protein; programs; promoter; receptor binding; recruit; regenerative; regenerative growth; stem; steroid hormone; success; tissue regeneration; tissue repair; tumor; wound; wound healing

PROJECT SUMMARY The fruit fly, Drosophila melanogaster, is a well-established model system to define conserved mechanisms that control proliferation, survival, invasiveness, and stem-like qualities of developing epithelia. A number of clinically significant oncogenes and tumor suppressors have been discovered and defined in flies (e.g. Yki/Yap1, Notch and archipelago/Fbw7), and multiple Drosophila tumor models are currently being used as discovery platforms to identify potential therapeutic compounds and cocktails. The Drosophila Taiman (Tai) and Yorkie (Yki) transcriptional co-activator proteins are the sole fly orthologs of the well-established human oncoproteins SRC-3 and Yap1. Tai is the prime coactivator for the ecdysone receptor (EcR), a functional homolog of nuclear hormone receptors, that controls many aspects of epithelial and stem cell development. Yki is the main transcriptional effector of the Hippo pathway. Our laboratory discovered that Yki and Tai physically associate on target gene promoters and that this Tai-Yki complex enables crosstalk between the Hippo and steroid hormone pathways during developmental and pathologic growth in Drosophila. The current project is focused on a role for the Tai-Yki axis in homeostatic regrowth of wing epithelium following wounding, and thus lies at the intersection of wound healing and cancer noted by Harold Dvorak’s prescient description of cancer as “wounds that do not heal”. I have found preliminary evidence that transient and local reduction in the ability of cells surrounding epithelial wounds to synthesize 20-hydroxyecdysone (20HE), the steroid ligand of EcR, inhibits wound repair and regeneration. In parallel, I have found that EcR transcriptional activity is also significantly induced within cells surrounding the wound. These data, in light of the Tai-Yki complex, could link 20HE/EcR steroid signaling to Yki/Yap1-driven growth at the site of the wound. Indeed, Tai supports expression of certain Yki pro-growth targets in uninjured discs that are upregulated during regeneration (e.g. dIlp8). Thus, I will test the hypothesis that the steroid hormone 20HE is required for imaginal wing disc regeneration through promoting transcriptional activity of Tai-dependent Yki targets. The Specific Aims of this project are: 1) test the requirement for 20HE biosynthesis in wing disc regeneration; 2) define the pattern of EcR activity in regenerating wing discs; 3) test the local requirement for the Tai-Yki axis in regenerative growth. In Aim 1, I will compare the extent of regeneration of wing discs depleted of 20HE biosynthesis genes to control discs, and determine the pattern of 20HE biosynthesis gene expression in regenerating imaginal wing discs. In Aim 2, I will place EcR activation into the context with other pathways activated during regeneration. In Aim 3, I will determine the effect of disrupting Tai-Yki association on the extent of regeneration and Tai-dependent Yki target expression in wing discs. Success in these studies will provide insight into how local hormone synthesis regulates homeostatic wound repair and, potentially, pathologic cancer growth.

项目摘要 果蝇果蝇果蝇是一个确定保守机制的良好模型系统 控制上皮亚的刺激，生存，侵入性和类似茎状的品质。 已经发现并定义了苍蝇（例如 YKI/YAP1，Notch和Ilcipelago/FBW7）以及多个果蝇肿瘤模型目前被用作 识别潜在的治疗化合物和鸡尾酒的发现平台。 和约克（YKI）转录共激活因子蛋白是公认的人类的唯一苍蝇直系同源物 癌蛋白SRC-3和YAP1。 核激素受体的同源物，它控制着上皮和干细胞发育的许多方面。 YKI是河马途径的主要转录效应子。 在靶基因启动子和太极拳复合物上的物理缔合使得可以在 果蝇发育和病理学中的河马和类固醇激素途径。 当前项目的重点是在机翼上皮的稳态再生中的台湾轴的角色 受伤后，因此位于Harold Dvorak的伤口愈合和癌症的交汇处 对癌症的有先见之明的描述为“无法愈合的伤口”。 以及局部修复周围细胞周围上皮伤口合成20-羟基甲虫的能力 （20HE）ECR的类固醇配体抑制了伤口修复和再生。 鉴于这些数据，转录活性也显着诱发了周围的WITIN细胞。 Tai-Yki复合物可以将20HE/ECR类固醇信号与YKI/YAP1驱动的生长联系起来。 实际上，太极支持在未受伤的光盘中表达某些YKI培养靶标的表达 再生（例如DILP8）。 想象中的翼圆盘再生通过促进tai依赖性YKI靶标的转录活性 该项目的特定目的是：1）测试机翼圆盘再生20个生物合成的需求； 定义再生翼盘中的ECR活性模式； 3） 在AIM 1中的再生生长，我将比较20人 生物合成基因控制椎间盘，并确定20HE生物合成基因表达的模式 在AIM 2中，我将ECR激活在其他小动物的情况下，将ECR激活 在AIM 3中激活。 翼盘的再生和TAI-末端YKI靶标表达的程度。 提供有关局部激素合成如何调节稳态伤口修复的信息 病理癌的生长。