REGULATION OF THE HUMAN CD34 GENE DURING HEMATOPOIESIS

这两个阶段的项目旨在提供科学的培训， 为了更好地了解造血功能， 干/祖细胞（HSPC）基因调控的研究。 调节CD 34的因子 CD 34在HSPC上特异性表达，并随着血液中CD 34的表达而下调。 细胞成熟。迄今为止，它是人类HSPC唯一明确的标志物。 CD 34+细胞可以重建所有细胞谱系的正常造血， 可能是多种白血病的起源细胞。此外，HSPC 是基因治疗的理想潜在靶点。先前的研究 适当调节细胞中CD 34表达的确定的控制元件 线然而，这些元素还有待于在正常的模型中进行研究。 干/祖细胞造血。 第一阶段将提供基础科学知识的发展， 实验室研究技能，通过一个程序，包括教学 培训和实验室研究。第一阶段研究的目标是 是为了证明控制CD 34的调节元件 在细胞系中的表达也调节其在正常细胞中的表达。 造血细胞使用CD 34启动子/增强子的表达研究 将在转基因小鼠和人CD 34+细胞中进行构建 正常造血的培养模型。 第二阶段将通过以下方式促进研究者独立性： 概念化和执行密集的研究计划的基础上， 第一阶段的结果。具体目标是分离最低限度的CD 34 将指导异源基因表达的调控元件， HSPC，并分离和表征参与HSPC的元素。 在造血分化过程中下调CD 34。构建体 将在我们的模型中进行设计和测试 系统. 这些研究有助于更好地了解HSPC基因 调节并导致对白血病发生和基因治疗的新见解。

This two phase project is designed to provide for the scientific training of the Investigator and to achieve a better understanding of hematopoietic stem/progenitor cell (HSPC) gene regulation through the study of the factors regulating CD34. CD34 is expressed specifically on HSPCs and is downregulated as blood cells mature. To date, it is the only well defined marker for human HSPCs. CD34+ cells can reconstitute normal hematopoiesis of all cell lineages and may be the cells of origin in a variety of leukemias. In addition, HSPCs are ideal potential targets for gene therapy. Previous studies have defined control elements which properly regulate CD34 expression in cell lines. However, these elements have yet to be studied in models of normal stem/progenitor cell hematopoiesis. Phase I will provide for the development of basic scientific knowledge and laboratory research skills through a program consisting of didactic training and active laboratory research. The goal of Phase I research will be to demonstrate that the regulatory elements which control CD34 expression in cell lines also regulate its expression in normal hematopoietic cells. Expression studies using CD34 promoter/enhancer constructs will be performed in both transgenic mouse and human CD34+ cell culture models of normal hematopoiesis. Phase II will promote Investigator independence through the conceptualization and execution of an intensive research plan based on Phase I results. The specific aims will be to isolate the minimal CD34 regulatory elements which will direct expression of a heterologous gene in HSPCs and to isolate and characterize the elements involved in the downregulation of CD34 during hematopoietic differentiation. Constructs containing these elements will be engineered and tested in our model systems. These studies should lead to a better understanding of HSPC gene regulation and lead to new insights into leukemogenesis and gene therapy.