Mechanisms controlling cell cycle exit upon terminal differentiation

终末分化时控制细胞周期退出的机制

• 批准号: 8206617
• 负责人: Laura A Buttitta
• 金额: $ 23.77万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206617
• 关键词: Address; Award; B-Lymphocytes; Biochemical; Biological Models; Cancer cell line; Cancerous; Candidate Disease Gene; Cell Culture System; Cell Culture Techniques; Cell Cycle; Cell Cycle Regulation; Cell Line; Cell division; Cells; Coupled; Development; Drosophila eye; Drosophila genus; Effector Cell; Eye; Future; Genes; Genetic; Genetic Epistasis; Genetic Screening; Goals; Grant; Human; In Vitro; Instruction; Malignant Neoplasms; Mammalian Cell; Mammals; Orthologous Gene; Pathway interactions; Phase; Phenotype; Post-Transcriptional Regulation; Process; Proliferating; RNA Interference; Research; Role; Signal Pathway; Signal Transduction; System; Testing; Tissues; Wing; Work; base; cancer therapy; experience; fly; insight; mature animal; novel; overexpression; prevent; tool

Terminal differentiation is often coupled with permanent exit from the cell cycle and represents the most common cellular state in adult animals. Yet it remains unclear how proliferation is blocked in differentiated tissues. The goal of this project is to determine how terminal differentiation signals impinge on the cell cycle machinery to induce a stable quiescent state and investigate how this state is disrupted in cancer. In the research proposed here, I use a combination of genetic and biochemical approaches in Drosophila and mammalian cells to delineate the conserved genetic pathways that control cell cycle exit. During the K99 phase of this grant, I investigated the redundant mechanisms that limit cycling in differentiated Drosophila tissues, obtained experience working with 2 different mammalian cell lines that can be induced to differentiate in vitro for future studies of cell cycle exit, and worked on a screen to identify genes that when overexpressed or inhibited by RNAi, de-regulate the cell cycle to cause ectopic proliferation in contexts of terminal differentiation. Such genes would normally be activated or inhibited upon terminal differentiation, but likely become de-regulated in cancers. During the ROO phase of the award, I will determine the precise mechanisms by which these genes regulate the cell cycle (Aim 1) and examine the roles of mammalian orthologs of these genes in normal and cancerous mammalian cells (Aim2). I will also investigate the signaling pathways by which these genes are regulated, using genetic epistasis tools in both Drosophila and mammalian cells (Aim 3). Such work will begin to delineate the conserved pathways connecting terminal differentiation signals with cell cycle controls, and determine how they can become de-regulated leading to cancer.

终末分化通常伴随着细胞周期的永久退出，代表了细胞周期中最重要的分化。 成年动物的常见细胞状态。然而，目前尚不清楚在分化的细胞中增殖是如何被阻止的。 组织中这个项目的目标是确定终末分化信号如何影响细胞周期 机器诱导稳定的静止状态，并研究这种状态如何在癌症中被破坏。在 在这里提出的研究中，我在果蝇中使用遗传和生物化学方法的组合， 哺乳动物细胞描绘控制细胞周期退出的保守遗传途径。在K99 在这个阶段，我研究了限制分化果蝇循环的冗余机制， 组织，获得了使用2种不同哺乳动物细胞系的工作经验，这些细胞系可以诱导 在体外分化为未来的研究细胞周期退出，并在屏幕上工作，以确定基因，当 过表达或被RNAi抑制，使细胞周期失调，从而导致异位增殖。 终末分化这些基因通常在终末分化时被激活或抑制，但 可能在癌症中失去控制。在ROO阶段的奖励，我将确定准确的 这些基因调控细胞周期的机制（目的1），并检查哺乳动物细胞的作用。 这些基因在正常和癌性哺乳动物细胞中的直系同源物（Aim2）。我也会调查 这些基因被调控的信号通路，在果蝇和 哺乳动物细胞（Aim 3）。这项工作将开始描绘连接终末细胞的保守通路。 细胞周期控制的分化信号，并确定它们如何变得失调，导致 癌