TEL-AML1 Transgenic Zebrafish Model of Human Leukemia

TEL-AML1 人类白血病转基因斑马鱼模型

• 批准号: 7498005
• 负责人: Hatim Sabaawy
• 金额: $ 16.3万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-19 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7498005
• 关键词: Acute Lymphocytic Leukemia; B-Cell Acute Lymphoblastic Leukemia; B-Lymphocytes; Biology; Cell Differentiation process; Child; Childhood Acute Lymphocytic Leukemia; Chimeric Proteins; Chromosomal translocation; Chromosome abnormality; Clinical; Clinical Data; Data; Development; Doctor of Medicine; Doctor of Philosophy; Embryo; Enhancers; Event; Fellowship; Fishes; Flow Cytometry; Fluorescence Microscopy; Gene Transfer; Gene Transfer Techniques; Genes; Genetic; Goals; Heavy-Chain Immunoglobulins; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; High Prevalence; Human; In Situ Hybridization; Incidence; Insertional Mutagenesis; K-Series Research Career Programs; Lymphoblastic Leukemia; Malignant - descriptor; Malignant Childhood Neoplasm; Modeling; Molecular Abnormality; Monocytic leukemia; Monozygotic Twinning; Monozygotic twins; Mus; Mutagenesis; Mutation; National Cancer Institute; Oncogenes; One-Step dentin bonding system; Pathway interactions; Phenotype; Pliability; RUNX1 gene; Reporter Genes; Research; Research Personnel; Research Project Grants; Resources; Retroviridae; Reverse Transcriptase Polymerase Chain Reaction; System; TEL-AML1 Fusion Protein; TEL-AML1 Oncogene; Testing; Tissues; Transgenic Mice; Transgenic Organisms; Transplantation; Western Blotting; Zebrafish; Zinc Fingers; anticancer research; base; career; design; drug development; fetal; histone acetyltransferase; in utero; knock-down; leukemia; mutant; novel; prenatal; programs; promoter; research study; response; retroviral-mediated; t(12; 21)(p13; q22); transcription factor; transgene expression; tumor

DESCRIPTION (provided by applicant): Immediate Goal: To generate TEL-AML1 transgenic zebrafish model and investigate the secondary genetic events that accentuate leukemia development. Career Development Goal: To develop an independent career in leukemia research. Research Project: The t(12;21) chromosomal translocation, which generates the TEL-AML1 fusion gene, is the most frequent structural genetic abnormality in childhood cancer, and is exclusively associated with precursor B-cell acute lymphoblastic leukemia (ALL). Evidence suggests that the TEL-AML1 translocation usually occurs In Utero during fetal hematopoiesis and constitutes an initiating or first-hit mutation that is necessary but insufficient for leukemia development. The aim of this proposal is to identify the secondary genetic events required for the development of TEL-AML1-induced leukemia. The two specific aims designed to test the central hypothesis that TEL-AML1 fusion protein requires secondary genetic events to induce leukemic transformation are: 1) To generate stable transgenic zebrafish lines expressing the TEL-AML1 fusion protein, and evaluate the effects of transgene expression on zebrafish hematopoiesis and leukemic development; 2) To develop a "second-hit" model in TEL-AML1 transgenic zebrafish by investigating the altered expression of the normal zebrafish TEL and histone acetyltransferase activity, both implicated in TEL-AML1 leukemic development, and in parallel, by initiating a retroviral insertional mutagenesis screen in transgenic zebrafish aimed at identifying cancer genes that synergize with the TEL-AML1 fusion protein in ALL development. Resources available to carry out these aims include our established TEL-AML1 transgenic founders and their progeny, a zebrafish mutant for histone acetyltransferase, and a retrovirus expressing zebrafish-specific reporter gene. The zebrafish system is uniquely suitable for these experiments in that it presents a vertebrate model with a versatile biology, genetic flexibility, conserved hematopoietic development, and the opportunity to conduct transgenesis and subsequent mutagenesis. The ability to perform a large-scale phenotype-driven mutational screen to identify cooperating events that accelerate leukemic development in the presence of chromosomal fusion proteins represents the unique advantage of this model. The identification of crucial genes in the leukemic pathway in these studies may identify novel targets for leukemia specific therapies. Although this application is for a K22 career development award, without an institutional affiliation, I believe that having a postdoctoral fellowship at the National Cancer Institute for three years prepared me to develop an independent cancer research career, and that the favorable response to this application would significantly facilitate this transition. My long-term goal is to identify the genetic events associated with the development of leukemia in children, and the ability to perform the proposed studies is one step towards this vital goal.

描述（由申请人提供）：直接目标：生成Tel-AML1转基因斑马鱼模型并研究突出白血病发育的次要遗传事件。职业发展目标：发展白血病研究的独立职业。研究项目：产生Tel-AML1融合基因的T（12; 21）染色体易位是儿童癌症中最常见的结构性遗传异常，并且与前体B细胞急性淋巴细胞性白血病（ALL）完全相关。有证据表明，Tel-AML1易位通常发生在胎儿造血期间的子宫内，并且构成了必要的启动或第一击突变，但不足以进行白血病发育。 该提案的目的是确定开发Tel-AML1引起的白血病所需的二级遗传事件。旨在测试中心假设的两个具体目的是，Tel-AML1融合蛋白需要次级遗传事件诱导白血病转化，是：1）生成稳定的转基因斑马鱼线，表达Tel-AML1融合蛋白，并评估转基因表达对跨因子表达的影响。斑马鱼造血和白血病发育； 2）通过研究正常斑马鱼TEL和组蛋白乙酰基转移酶活性的变化，在Tel-AML1转基因斑马鱼中开发“第二次打击”模型，既涉及Tel-AML1白血病发育转基因斑马鱼中的诱变筛查旨在鉴定所有发育中与Tel-AML1融合蛋白协同作用的癌基因。可以实现这些目标的资源包括我们已建立的Tel-AML1转基因创始人及其后代，一种用于组蛋白乙酰转移酶的斑马鱼突变体以及表达斑马鱼特异性记者基因的逆转录病毒。 斑马鱼系统非常适合这些实验，因为它提出了具有多功能生物学，遗传柔韧性，保守的造血发展以及进行转基因和随后诱变的机会。执行大规模表型驱动的突变筛选的能力，以确定在存在染色体融合蛋白的情况下加速白血病发育的合作事件，这代表了该模型的独特优势。这些研究中白血病途径中关键基因的鉴定可以鉴定白血病特定疗法的新靶标。 尽管此申请是K22职业发展奖，但没有机构隶属关系，但我相信在国家癌症研究所获得博士后奖学金三年，我准备发展独立的癌症研究职业，对此申请的有利回应将会显着促进了这一过渡。我的长期目标是确定与儿童白血病发展相关的遗传事件，进行拟议的研究的能力是朝着这一重要目标迈出的一步。