ERYTHROPOIETIN & REGULATION OF ERYTHROPOIESIS

促红细胞生成素

• 批准号: 3225409
• 负责人: SANFORD B KRANTZ
• 金额: $ 24.3万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1974
• 资助国家: 美国
• 起止时间: 1974-10-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3225409
• 关键词: Friend virus; anemia; autoradiography; colony stimulating factor; erythroid stem cell; erythropoiesis; erythropoietin; glycosylation; human subject; insulinlike growth factor; interleukin 1; interleukin 3; laboratory mouse; membrane proteins; phosphorylation; protein structure function; radiotracer; receptor; receptor expression; tissue /cell culture; tumor necrosis factor alpha

The aim of this research is to enhance understanding of the regulation of erythropoiesis and the application of this knowledge to clinical anemias and polycythemias. We have purified human colony forming units-erythroid (CFU-E) and have identified erythropoietin (EPO) receptors on these cells. We now propose to purify blood burst-forming units-erythroid (BFU-E) and determine the time course of the evolution of EPO receptors during BFU-E development and differentiation. We will also determine the effect of interleukin-3 (IL-3) granulocyte colony-stimulating factor, granulocytemacorphage colony-stimulating factor, and insulin-like growth factor-I (IGF-I) on down- or up-modulation of EPO receptors. We have also found that while tumor necrosis factor (TNF) and IL-1 depress normal human bone marrow CFU-E development, purified CFU-E are not inhibited by these factors. Thus, TNF and IL-1 are apparently acting on accessory cells to produce a factor that is released and inhibits CFU-E. We will determine the identity of these intermediate accessory cells, purify the factor that is being released, and measure the effect of this factor on human CFU-E EPO receptors to determine if TNF and/or IL-1 down- module EPO receptors as a mechanism for producing the anemia. We will also further characterize the EPO receptor during erythroid differentiation. A heterobifunctional, photoreactive, cleavable, iodinated cross-linker will provide radioactive EPO receptors which will be characterized for molecular size, isoelectric point, glycosylation and phosphorylation capacity. The EPO receptor gene will be obtained and will be used to measure the effect of EPO and IGF-I on EPO receptor mRNA in highly purified murine CFU-E as they differentiate. These studies will enhance our knowledge of the events that are controlled by EPO and they will enhance our under-standing of the anemia of chronic disorders. They should also provide a firm base of information for further understanding a wide variety of additional clinical anemia and polycythemias that may occur through the malfunction of normal EPO control mechanisms.

本研究的目的是加深对监管的理解 红细胞生成以及该知识在临床贫血中的应用 和红细胞增多症。 我们纯化了人类集落形成单位-红细胞 (CFU-E) 并已鉴定出这些细胞上的促红细胞生成素 (EPO) 受体。 我们现在建议纯化血爆发形成单位-红细胞（BFU-E）和 确定 BFU-E 期间 EPO 受体进化的时间过程 发展和分化。 我们还将确定效果 白细胞介素 3 (IL-3) 粒细胞集落刺激因子， 粒细胞巨噬细胞集落刺激因子和胰岛素样生长 I 因子 (IGF-I) 对 EPO 受体的下调或上调。 我们还发现，虽然肿瘤坏死因子 (TNF) 和 IL-1 会抑制 正常人骨髓CFU-E发育，纯化CFU-E则不 受这些因素的抑制。 因此，TNF 和 IL-1 显然作用于 辅助细胞产生释放并抑制 CFU-E 的因子。 我们将确定这些中间辅助细胞的身份， 纯化正在释放的因子，并测量其效果 影响人 CFU-E EPO 受体的因素，以确定 TNF 和/或 IL-1 是否下调 模块 EPO 受体作为产生贫血的机制。 我们还将进一步表征红系期间的 EPO 受体 差异化。 异双功能、光反应性、可裂解、碘化 交联剂将提供放射性 EPO 受体，其将 表征分子大小、等电点、糖基化和 磷酸化能力。 获得EPO受体基因并将 用于测定 EPO 和 IGF-I 对 EPO 受体 mRNA 的影响 高度纯化的鼠 CFU-E，因为它们分化。 这些研究将 增强我们对 EPO 控制的事件的了解，它们 将增强我们对慢性疾病贫血的了解。 他们 还应为进一步了解 可能发生的各种其他临床贫血和红细胞增多症 由于正常 EPO 控制机制发生故障。