E2F3: a suppressor of centrosome amplification

E2F3：中心体扩增的抑制因子

• 批准号: 7228923
• 负责人: Harold I Saavedra
• 金额: $ 15.71万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7228923
• 关键词: Alleles; Aneuploidy; Cell Cycle; Cell Cycle Progression; Centrosome; Characteristics; Code; Development; Disruption; Doctor of Philosophy; E2F3 protein; Embryo; Epithelial Cells; Equilibrium; Fellowship; Fibroblasts; Frequencies; Genes; Genome; Genomic Instability; Goals; Growth; Human; Individual; Institution; Knock-out; Laboratories; Lead; Malignant Neoplasms; Mammary Tumorigenesis; Mammary gland; Mentorship; Molecular; Mus; Pathway interactions; Play; Rate; Repression; Role; Series; Transcript; Transcriptional Activation; Transgenic Mice; Trimethoprim-Sulfamethoxazole; Work; carcinogenesis; in vivo; in vivo Model; knock-down; transcription factor; tumor; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): The overall goal of this K01 proposal is to allow Harold I. Saavedra, Ph.D., under the mentorship of Gustavo Leone, Ph.D. to transition from a postdoctoral fellowship to the establishment of his own independent laboratory in an academic institution. The study of the Rb/E2F pathway is crucial to the understanding of cancer, since components of this pathway are de-regulated in almost all human tumors. One of the central components of the Rb/E2F pathway is the E2F3 transcription factor, which plays a key role in cell cycle progression. The E2F3 locus codes for two transcripts, E2F3a and E2F3b, whose individual functions are presently unknown. We show that gene knock-out of E2F3, which included knocking-out of E2F3a and E2F3b, leads to slower growth rates, centrosome amplification and aneuploidy. Our hypothesis is that the uncoupling of the centrosome cycle and the cell cycle, triggered by the loss of E2F3 function, may ultimately lead to cancer. To explore this hypothesis we propose the following specific aims: 1. To unravel the individual contributions of E2F3a and E2F3b towards ceil cycle control, suppression of centrosome amplification and aneuploidy. SiRNAs will be introduced into mouse embryonic fibroblasts to knock-down the expression of E2F3a and E2F3b. We will determine the growth characteristics of those knock-downs, and their individual contributions towards centrosome amplification and aneuploidy. 2. To explore, using a centrin-GFP transgenic mouse, whether loss of E2F3 leads to centrosome amplification in vivo. We will introduce a conditional allele of E2F3 into centrin-GFP mice. Conditional deletion of E2F3 in mammary epithelial cells will be achieved by crossing those mice with WAP-CRE mice. Frequencies of centrosome amplification will be calculated from sections of those mammary glands. 3. To explore the potential role of the myclE2F pathway in centrosome amplification and mammary carcinogenesis. We will generate WAP-myc/centrin-GFP mice and determine progression from expression of myc to centrosome amplification, aneuploidy and tumorigenesis. We will also explore whether loss of E2F3 cooperates with myc to accelerate mammary tumorigenesis. The results obtained from this K01 proposal will be invaluable to the understanding of the relationship between centrosome amplification, aneuploidy and carcinogenesis in vivo, and to whether genomic instability is a cause or a consequence of tumorigenesis.

描述（由申请人提供）：该K01提案的总体目标是允许Harold I. Saavedra，Ph.D.，在Gustavo Leone博士的指导下。从博士后研究金过渡到在学术机构中建立自己的独立实验室。 RB/E2F途径的研究对于对癌症的理解至关重要，因为该途径的组成部分在几乎所有人类肿瘤中都被驱散。 RB/E2F途径的中心成分之一是E2F3转录因子，在细胞周期进程中起关键作用。 E2F3基因座代码为两个转录本E2F3A和E2F3B，其各个函数目前未知。我们表明，E2F3的基因敲除包括敲除E2F3A和E2F3B的敲除，导致生长速度较慢，中心体扩增和非整倍性。我们的假设是，由E2F3功能的丧失触发了中心体周期和细胞周期的解偶，最终可能导致癌症。为了探讨这一假设，我们提出了以下特定目的：1。阐明E2F3A和E2F3B对CEIL循环控制的个体贡献，抑制中心体扩增和非整倍性。 siRNA将被引入小鼠胚胎成纤维细胞中，以敲低E2F3A和E2F3B的表达。我们将确定这些敲击的生长特征，以及它们对中心体扩增和非整倍性的个人贡献。 2。探索，使用中心蛋白-GFP转基因小鼠，E2F3的损失是否导致体内中心体扩增。我们将将E2F3的条件等位基因引入中心蛋白GFP小鼠。通过将这些小鼠与WAP-CRE小鼠交叉，可以实现乳腺上皮细胞中E2F3的条件缺失。中心体扩增的频率将根据这些乳腺的部分计算。 3。探索Mycle2F途径在中心体扩增和乳腺癌发生中的潜在作用。我们将生成WAP-MYC/Centrin-GFP小鼠，并确定从MYC表达到中心体扩增，非整倍性和肿瘤发生的发展。我们还将探索E2F3的损失是否与MYC合作以加速乳腺肿瘤。从该K01提案获得的结果对于理解体内中心体扩增，非整倍性和癌变之间的关系以及基因组不稳定性是肿瘤发生的原因还是后果是无价的。