HEMATOPOIETIC STEM CELL DIFFERENTIATION IN HUMAN BLOOD

人类血液中的造血干细胞分化

• 批准号: 6123167
• 负责人: BARBARA A. MILLER
• 金额: $ 3.26万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6123167
• 关键词: biological signal transduction; bone marrow; calcium channel; cell cycle; cell differentiation; cell free system; clinical research; electrophysiology; erythroid stem cell; erythropoietin; growth factor; hematopoiesis; human subject; membrane channels; transcription factor; voltage /patch clamp

The long term goal of studies proposed here is to determine the signal transduction mechanisms through which growth factors stimulate hematopoietic proliferation and differentiation. The major goal of this project is to understand the mechanisms through which erythropoietin (Epo) regulates ion channels during erythroid differentiation and to determine the functional role of calcium influx in erythropoiesis. Blood will be drawn and bone marrow aspirates will be performed in the CRC on healthy volunteers. The following specific aims will be addressed: Specific Aim 1: Identification of the signaling mechanism through which Epo regulates calcium channels. (A) Here we will determine the Epo signaling pathways which link Jak2 to calcium channel activation. Involvement of STAT, Ras, or the IRS-2/PI 3 kinase pathways will be examined using microinjection of single BFU-E derived erythroblasts and quantitative fluorescence microscopy coupled digital video imaging. (B) We will determine the domains of erythropoietin receptor required for Epo modulation of calcium channel opening. Specific Aim 2:Determination of the role of [Cai] in regulation of transcription factor activation in erythropoiesis. The function of the NF-kB and bHLH transcription factors and the c-Jun N-terminal kinases (JNK) are modulated by calcium. We will determine if the amplitude or duration of the Ca++ response in erythroid cells affects NF-kB transcription factor activation, or if calcium/calmodulin levels influence DNA binding of bHLH proteins, particularly SCL. We will also determine the effect of [Cai] on JNLK activation on the role of PI 3-kinase in this pathway.

这里提出的研究的长期目标是确定生长因子刺激造血增殖和分化的信号转导机制。 该项目的主要目的是了解红细胞生成素（EPO）在红细胞分化过程中调节离子通道的机制，并确定钙涌入在红细胞生成中的功能作用。 将吸收血液，并将在健康志愿者的CRC中进行骨髓抽吸物。 将解决以下特定目的：具体目标1：识别EPO调节钙通道的信号传导机制。 （a）在这里，我们将确定将JAK2与钙通道激活联系起来的EPO信号通路。将使用单个BFU-E衍生的生成颗粒细胞和定量荧光显微镜耦合数字视频成像的微注射来检查STAT，RAS或IRS-2/PI 3激酶途径的参与。 （b）我们将确定钙通道开放的EPO调节所需的红细胞生成素受体的结构域。 具体目的2：确定[CAI]在促红细胞生成中转录因子激活中的作用。 NF-KB和BHLH转录因子以及C-JUN N末端激酶（JNK）的功能通过钙调节。 我们将确定红细胞细胞中Ca ++反应的幅度或持续时间是否会影响NF-KB转录因子激活，或者钙/钙调蛋白水平是否影响BHLH蛋白（尤其是SCL）的DNA结合。 我们还将确定[CAI]对JNLK激活对PI 3-激酶在该途径中的作用的影响。