Elucidating the role of Ftz-f1 as a mediator of steroid hormone-dependent activity in stem cells and their progeny

阐明 Ftz-f1 作为干细胞及其后代中类固醇激素依赖性活性调节剂的作用

• 批准号: 9021098
• 负责人: Elizabeth Tweedie Ables
• 金额: $ 43.49万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9021098
• 关键词: Address; Alleles; Anterior; Biological; Biological Assay; Cell Count; Cell Cycle; Cell Cycle Regulation; Cell Differentiation process; Cell Maintenance; Cell Proliferation; Cell Therapy; Cell physiology; Cells; Childhood; Clinical; Cues; Data; Detection; Disease; Drosophila genus; Drosophila melanogaster; Ecdysone; Embryo; Environment; Equilibrium; Experimental Genetics; Future; Genes; Genetic; Graafian Follicles; Growth and Development function; Health; Homeostasis; Hormonal; Hormones; Human; Individual; Insecta; Left; Location; Maintenance; Measures; Mediating; Mediator of activation protein; Methods; Molecular; Natural regeneration; Nuclear; Nuclear Hormone Receptors; Oocytes; Oogenesis; Orphan; Ovarian; Ovary; Pathway interactions; Physiological; Physiology; Population Sizes; Pregnancy; Production; Property; Receptor Signaling; Regenerative Medicine; Reproduction; Role; Signal Pathway; Signal Transduction; Somatic Cell; Staging; Stem cells; Steroids; Techniques; Technology; Testing; Tissues; cancer therapy; cell type; competence factor; daughter cell; ecdysone receptor; germline stem cells; in vivo; insight; interest; knock-down; meetings; mutant; novel; programs; regenerative therapy; repaired; research study; response; self-renewal; stem; stem cell fate; stem cell population; steroid hormone; tool; transcription factor

PROJECT SUMMARY Recent advances in cell reprogramming have raised considerable interest in the use of stem cells for regenerative therapies to replace damaged or lost cells. Stem cells integrate a variety of intrinsic and extrinsic cues to maintain their fate and proliferative capacity. Importantly, these cues include steroid hormones, which fluctuate during childhood growth and development, pregnancy, and disease, and nuclear hormone receptors, which receive steroid signals to regulate physiological homeostasis. A basic understanding of how stem cells respond to changes in human physiology is therefore a necessary prerequisite to the increased use of stem cells in clinical therapy. Like other nuclear hormone receptor signaling pathways, the insect steroid hormone ecdysone is known to elicit context-dependent cellular responses; however, it is largely unknown how these responses are achieved. The nuclear hormone receptor Ftz-f1 is an ecdysone target, but it is also required for the maximal transcriptional response of other ecdysone targets (namely, E74 and E75) in embryonic and larval tissues. This suggests that Ftz-f1 activity primes some cells to respond to ecdysone by up- regulating a specific transcriptional pathway. Despite the identification of mutant alleles nearly 25 years ago, no studies to date have addressed the function of Ftz-f1 in the ovary. Our studies will test the central hypothesis that Ftz-f1 acts as a competence factor for ecdysone signaling in the ovary. Germline stem cells (GSCs) in the Drosophila melanogaster ovary are well-suited for these studies, due to their easily accessible anatomical location, sensitivity to steroid hormones, and the availability of genetic and experimental techniques. Our studies will not only reveal important new information about the function of ftz-f1 in stem cells and reproduction, but can also serve as a novel paradigm for the study of context-dependent steroid hormone signaling in vivo. In my new independent lab, we will utilize Drosophila genetic mutants and in vivo cell-specific gene knockdown to investigate whether and how ftz-f1 regulates GSCs and their progeny, and test whether ftz-f1 primes specific cells for ecdysone signaling. In Aim 1, we will examine the role of ftz-f1 in the differentiation of GSCs and their progeny. In Aim 2, we will investigate whether ftz-f1 controls the cell cycle in GSCs and their progeny. In Aim 3, we will use genetic interaction experiments to determine whether ftz-f1 functionally interacts with ecdysone signaling. Given the similarity between Drosophila and human steroid hormone signaling, our study will help elucidate how nuclear hormone receptors differentially regulate context-dependent transcriptional pathways in vivo to control stem cell function.

项目摘要 细胞重编程的最新进展引起了人们对使用茎的极大兴趣 细胞用于再生疗法以替代受损或丢失的细胞。干细胞整合了多种 保持其命运和增殖能力的内在和外在提示。重要的是，这些提示 包括类固醇激素，在儿童期生长和发育，怀孕和 疾病和核激素受体，它们接收类固醇信号以调节生理 稳态。对干细胞如何应对人类生理变化的基本理解 因此，这是增加干细胞在临床治疗中增加使用的必要先决条件。喜欢 其他核激素受体信号传导途径，已知昆虫类固醇激素ecdysone已知 引起依赖上下文的细胞反应；但是，这在很大程度上尚不清楚这些响应是如何的 成就了。核激素受体FTZ-F1是ecdysone靶标，但也需要 胚胎和 幼虫组织。这表明FTZ-F1活性素一些细胞通过上升来响应Ecdysone 调节特定的转录途径。尽管鉴定了近25年的突变等位基因 以前，迄今为止尚无研究解决卵巢中FTZ-F1的功能。我们的研究将测试 FTZ-F1是卵巢中ecdysone信号传导的能力因素的中心假设。 果蝇中的种系干细胞（GSC）非常适合这些研究 到他们易于访问的解剖位置，对类固醇激素的敏感性以及可用性 遗传和实验技术。我们的研究不仅会揭示有关的重要新信息 FTZ-F1在干细胞和繁殖中的功能，但也可以作为新的范式 在体内依赖上下文依赖性类固醇激素信号传导的研究。在我的新独立实验室中，我们将使用 果蝇基因突变体和体内细胞特异性基因敲低，以研究是否以及如何 FTZ-F1调节GSC及其后代，并测试FTZ-F1特定细胞的Ecdysone 信号。在AIM 1中，我们将研究FTZ-F1在GSC及其分化中的作用 后代。在AIM 2中，我们将研究FTZ-F1是否控制GSC中的细胞周期 后代。在AIM 3中，我们将使用遗传相互作用实验来确定FTZ-F1是否是否 在功能上与ecdysone信号相互作用。鉴于果蝇和人类之间的相似性 类固醇激素信号传导，我们的研究将有助于阐明核激素受体的差异性 在体内调节上下文依赖性转录途径以控制干细胞功能。