Novel molecular targets of RUNX in hematopoietic stem/progenitor cells

RUNX在造血干/祖细胞中的新分子靶点

• 批准号: 7597128
• 负责人: Shinobu Matsuura
• 金额: $ 5.17万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7597128
• 关键词: Acute Myelocytic Leukemia; Adult; Affect; Binding Sites; Blood Cells; Bone Marrow Cells; Cell Line; Cell physiology; Cells; Chromosomal translocation; DNA; DNA Binding; Data; Development; Disease; Dysmyelopoietic Syndromes; Embryo; Gene Expression; Gene Expression Alteration; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Human; In Vitro; Knock-out; Knockout Mice; Life; Maintenance; Malignant - descriptor; Methods; Microarray Analysis; Modeling; Molecular; Molecular Target; Mus; Mutate; Mutation; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Proliferating; Proteins; RUNX1 gene; Regulation; Regulator Genes; Research; Role; Stem cells; Syndrome; Techniques; Testing; Therapeutic; chromatin immunoprecipitation; gene discovery; genome-wide; in vitro Model; in vivo; insight; knockout gene; leukemia; mouse model; novel; simulation; stem; transcription factor

DESCRIPTION (provided by applicant): The long-term objective of this research is to clarify the genetic mechanisms by which hematopoietic stem cells proliferate and differentiate into mature blood cells. More specifically, this project will focus on the Runxl transcription factor in hematopoietic stem cell function. Runxl is a DNA binding transcription factor. However, its critical target genes in hematopoiesis are not known. Chromosomal translocations and mutations of Runxl are involved in the pathogenesis of leukemia and myelodysplastic syndrome. Runxl gene dysruption in mice causes embryonic lethality due to complete lack of definitive hematopoiesis. Conditional knockout of this gene in adult life causes perturbation of normal proliferation and differentiation in various hematopoietic lineages. This proposal will test a hypothesis that Runxl directly regulates the expression of a group of critical genes involved in the maintenance and differentiation of hematopoietic stem cells. This project will proceed in three steps. In Specific Aim 1, the mouse multipotent hematopoietic stem cell line EML, which is a good in vitro model of normal hematopoiesis, will be used for ChIP on chip analysis. ChIP on chip analysis combines chromatin immunoprecipitation and DNA tiling microarray analysis, and is a powerful technique for genome-wide search for transcription factor target genes. In Specific Aim 2, expression microarray analysis will be performed on hematopoietic stem cells from the Runxl conditional knockout mouse. The transcriptional profile of these cells will reveal the genes whose expression is controlled by RUNX1. The role of Runxl in illness will then be approached throuh the transcriptional profile of the Runxl (Ala224fsTer228) mutation, found in a patient with MDS/AML. In Specific Aim 3, the data obtained in Specific Aims 1 and 2 will be combined to picture the network of genes that are under direct control of the Runxl transcription factor. The major target genes and the downstream pathways affected by this gene will be characterized focusing on hematopoietic stem cell function. The most significant genes will be selected and tested for their significance in the hematopoietic system, using both in vitro and in vivo approaches. Normal hematopoiesis is a result of controlled expression of various genes in the hematopoietic stem cell. Runxl is a master regulator of gene expression in hematopoietic stem cells. Despite intense research in the field, the most critical Runxl target genes for normal hematopoiesis is still unknown. The goal of this project is to discover the genes indispensable for normal hematopoiesis. Understanding of the precise molecular mechanisms that control hematopoiesis is essential for the development of better therapeutic drugs for hematopoietic disorders.

描述（由申请人提供）：这项研究的长期目标是阐明造血干细胞增殖并分化为成熟血细胞的遗传机制。更具体地说，该项目将重点放在造血干细胞功能中的Runxl转录因子上。 RunXl是DNA结合转录因子。但是，其造血中的关键靶基因尚不清楚。 Runxl的染色体易位和突变参与白血病和骨髓增生综合征的发病机理。小鼠中的Runxl基因发病障碍会导致由于完全缺乏确定的造血作用而导致胚胎致死性。该基因在成人生活中的有条件敲除导致各种造血谱系中正常增殖和分化的扰动。该提案将检验一个假设，即RUNXL直接调节与造血干细胞维持和分化有关的一组关键基因的表达。该项目将分三个步骤进行。在特定的目标1中，小鼠多能造血干细胞系EML（这是正常造血的体外模型）将用于芯片分析。芯片分析中的芯片结合了染色质免疫沉淀和DNA平铺微阵列分析，是全基因组搜索转录因子靶基因的强大技术。在特定的目标2中，将从Runxl条件基因敲除小鼠对造血干细胞上进行表达微阵列分析。这些细胞的转录谱将揭示其表达受Runx1控制的基因。然后，将通过在MDS/AML患者中发现Runxl（Ala224fster228）突变的转录谱（ALA224FSUR228）突变的转录曲线来接触Runxl的作用。在特定目标3中，将组合在特定目标1和2中获得的数据，以描绘在Runxl转录因子直接控制基因网络中。该基因影响的主要靶基因和下游途径将被特征在于造血干细胞功能。最重要的基因将使用体外和体内方法选择并测试其在造血系统中的重要性。正常造血是造血干细胞中各种基因的控制表达的结果。 RunXl是造血干细胞中基因表达的主要调节剂。尽管在该领域进行了激烈的研究，但正常造血的最关键的RUNXL靶基因仍然未知。该项目的目的是发现正常造血必不可少的基因。了解控制造血的精确分子机制对于开发更好的造血疾病治疗药物至关重要。