REGULATION OF THE HUMAN CD34 GENE DURING HEMATOPOIESIS

人类 CD34 基因在造血过程中的调控

• 批准号: 2134288
• 负责人: IRA A OLIFF
• 金额: $ 7.88万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-30 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2134288
• 关键词: CD antigens; biological models; cell differentiation; cell growth regulation; clone cells; gene expression; genetic enhancer element; genetic promoter element; genetic regulation; genetically modified animals; hematopoiesis; hematopoietic stem cells; human subject; laboratory mouse; membrane proteins; surface antigens; transfection

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.

这两个阶段的项目旨在提供科学培训 研究者的意见并更好地了解造血系统 通过研究干/祖细胞 (HSPC) 基因调控 CD34 的调节因子。 CD34 在 HSPC 上特异性表达，并随着血液的变化而下调 细胞成熟。迄今为止，它是唯一明确的人类 HSPC 标记。 CD34+细胞可以重建所有细胞谱系的正常造血功能 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标。之前的研究有 确定的控制元件可以正确调节细胞中的 CD34 表达 线。然而，这些元素还有待在正常模型中进行研究。 干/祖细胞造血。 第一阶段将提供基础科学知识的发展 通过包括教学在内的项目培养实验室研究技能 培训和积极的实验室研究。第一阶段研究的目标是 证明控制 CD34 的调控元件 细胞系中的表达也调节其在正常情况下的表达 造血细胞。使用 CD34 启动子/增强子进行表达研究 将在转基因小鼠和人类 CD34+ 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的独立性： 基于此的深入研究计划的概念化和执行 第一阶段结果。具体目标是分离最小的 CD34 指导异源基因表达的调控元件 HSPC 并分离和表征涉及的元素 造血分化过程中 CD34 的下调。构造 包含这些元素将在我们的模型中进行设计和测试 系统。 这些研究应该有助于更好地了解 HSPC 基因 调控并带来对白血病发生和基因治疗的新见解。