THE ROLE OF FLI1 IN CELLULAR DIFFERENTIATION AND DEVELOPMENT

FLI1 在细胞分化和发育中的作用

• 批准号: 6340782
• 负责人: Dennis K Watson
• 金额: $ 15.41万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6340782
• 关键词: cell differentiation; cell growth regulation; cell proliferation; embryonic stem cell; erythroleukemia; gene deletion mutation; gene expression; gene targeting; genetic regulation; genetically modified animals; laboratory mouse; megakaryocytes; molecular site; neoplasm /cancer genetics; neoplastic cell; phenotype; protein structure function; tissue /cell culture; transcription factor

The ETS family of transcription factors has been implicated in the development of leukemias, lymphomas and sarcomas. We have previously identified a member of this family, FLI1, whose retroviral activation results in the development of hematological tumors. The FLI1 gene is oncogenically activated in the transformed cells of patients with Ewing's sarcoma by the t(11;22) translocation. The long range goal of this research is to understand the function of FLI1 in the regulation of cellular differentiation and development and how dysregulated FLI1 can lead to the development of disease. Our preliminary data indicate that FLI1 expression can promote differentiation of leukemic cells towards the megakaryocytic lineage. In addition, we have generated targeted cell lines that carry a mutant allele for the Fli1 gene. We propose to use these in vitro and in vivo systems to examine the consequences of over- and under-expression of FLI1. Ultimately, this will help us identify relevant target genes controlled by this member of the ETS gene family. We hypothesize that FLI1 functionally regulates genes that are critical for cellular differentiation, specifically genes encoding cell-specific proteins and genes that encode proteins that control cellular proliferation. The specific aims of this project are to: (1) define the role of FLI1 in the differentiation of human erythroleukemia cells (K562) by deletion analysis of FLI1; identify the domains required for its biological effects; to determine if ERG can functionally substitute for FLI1; (2) identify FLI1 target genes associated with promotion of cellular differentiation and regulation of cell growth by analysis of gene expression in an inducible system; and (3) to define the role of FLi1 in development by disruption of Fli1 gene in ES cells and in mice by homologous recombination; characterize the pathology of the mutant mice; including hematopoietic cell lineage profile analysis. The experiments proposed will provide important understanding into the mechanisms that control lineage selection during hematopoiesis and cellular proliferation, and insights into oncogenesis caused by the disruption of these controls.

转录因子的ETS家族已经被牵连在 白血病、淋巴瘤和肉瘤的发展。我们先前已经 发现了这个家族的一个成员，FLI 1，其逆转录病毒激活 导致血液肿瘤的发展。FLI 1基因是 在尤因病患者的转化细胞中致癌激活 t（11;22）易位的肉瘤。长期目标是 研究的目的是了解FLI 1在调节 细胞分化和发育以及FLI 1失调如何 导致疾病的发展。 我们的初步数据表明，FLI 1表达可以促进 白血病细胞向巨核细胞谱系的分化。在 此外，我们已经产生了携带突变等位基因的靶向细胞系， Fli 1基因。我们建议使用这些体外和体内系统， 研究FLI 1过度表达和表达不足的后果。 最终，这将帮助我们确定相关的靶基因控制的 这个ETS基因家族的成员我们假设FLI 1在功能上 调节对细胞分化至关重要的基因， 特别是编码细胞特异性蛋白质的基因和编码 控制细胞增殖的蛋白质。具体目标是 项目是：（1）确定FLI 1在区分 人红白血病细胞（K562）FLI 1缺失分析;鉴定 其生物学效应所需的结构域;以确定ERG是否可以 FLI 1的功能替代;（2）鉴定FLI 1靶基因 与促进细胞分化和调节 通过分析诱导系统中的基因表达的细胞生长;和（3） 通过在ES细胞中破坏Fli 1基因来确定Fli 1在发育中的作用 细胞和小鼠中通过同源重组;表征病理学 包括造血细胞谱系分析。 提出的实验将提供重要的理解， 造血过程中控制谱系选择的机制， 细胞增殖，并深入了解肿瘤发生引起的 破坏这些控制。