STK-1--A HUMAN HEMATOPOIETIC GROWTH FACTOR RECEPTOR

STK-1--人类造血生长因子受体

• 批准号: 2226552
• 负责人: DONALD SMALL
• 金额: $ 11.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2226552
• 关键词: antibody; biological signal transduction; cell differentiation; cell growth regulation; cellular pathology; clinical chemistry; flow cytometry; genetic regulation; genetic regulatory element; growth factor receptors; hematopoiesis; hematopoietic growth factor; hematopoietic stem cells; molecular cloning; neoplastic transformation; northern blottings; polymerase chain reaction; protein tyrosine kinase; receptor expression; reporter genes; tissue /cell culture; western blottings

Our long-term objectives for this project are to obtain an understanding of the control mechanisms involved in regulating human lymphohematopoietic stem (LHSC) and progenitor cells. This is an important area of research because of the potential clinical impact of being able to manipulate this process. We have taken the approach that receptor tyrosine kinases are likely to be important in regulating the cell numbers or the development of these cells. This premise is based on the accumulated evidence of their role in both hematopoietic and nonhematopoietic cells. We have successfully cloned the cDNA for a human receptor tyrosine kinase, STK-1, that is most closely related to c-fms and c-kit, both growth factor receptors that are involved in hematopoiesis. STK-1 was cloned from a population of human bone marrow cells that are greatly enriched for LHSC and progenitor cells. Initial studies indicate that the expression of STK-1 is restricted to this "young" fraction of bone marrow. The mouse homolog of this receptor was cloned recently and was shown to have similarly restricted pattern of expression. The receptor is therefore of interest because of the potential insights it may give us into the control of hematopoiesis. We have proposed a series of experiments for which most of the reagents are already available as an outcome of our preliminary work. We will carefully examine the expression of STK-1 in subfractionated human marrow by RT-PCR to determine in which cells STK-1 is likely to play a role. The fractions tested will include some of the most highly purified LHSC, to give us an indication of whether the receptor is expressed on these cells. Antibodies to the receptor will allow us to confirm these results by FACS analysis and may enable the further purification of these cells. We will investigate the regulation of the gene coding for STK-1. We will first complete the cloning of the region containing the gene. We will search for cis-acting sequences that confer expression by use of a reporter gene assay. Thus insight may be gained into the control of other genes with expression similarly restricted to "young" hematopoietic cells. We will analyze the pathway by which STK-1 signal transduction takes place. We will look at the targets of signal transduction already identified for other receptors and hope to find some novel proteins as well. Lastly, we will test the hypothesis that this receptor is involved in malignancy or other disease processes involving the hematopoietic system. The finding that STK-1 is expressed in several leukemic cell lines may be an indication that it is somehow involved. RNA and DNA samples from patients afflicted with these diseases will be tested over-expression or altered expression of the receptor.

我们对这个项目的长期目标是了解 参与调节人类行为的控制机制 淋巴造血干细胞（LHSC）和祖细胞。 这是一 重要的研究领域，因为潜在的临床影响 能够操纵这个过程。 我们采取的方法是， 受体酪氨酸激酶可能在调节 细胞数量或这些细胞的发育。 这个前提是基于 根据其在造血和 非造血细胞 我们已经成功地克隆了人酪氨酸受体的cDNA 激酶STK-1与c-fms和c-kit的关系最为密切， 与造血有关的生长因子受体 STK-1是 从人类骨髓细胞中克隆出来的， 富集LHSC和祖细胞。 初步研究表明， STK-1的表达仅限于骨的这一“年轻”部分 骨髓 最近克隆了该受体的小鼠同源物， 显示具有类似的受限表达模式。 受体 因为它可能提供潜在的见解， 控制造血系统 我们提出了一系列实验，其中大多数试剂 已经作为我们初步工作的成果提供。 我们将 仔细检查STK-1在亚分级人骨髓中的表达 通过RT-PCR来确定STK-1可能在哪些细胞中发挥作用。 测试的级分将包括一些最高度纯化的LHSC， 来给我们一个指示， 细胞 受体的抗体将使我们能够证实这些结果 通过流式细胞仪分析，可以进一步纯化这些细胞。 我们将研究编码STK-1的基因的调控。 我们将 首先完成含有该基因的区域的克隆。 我们将 通过使用一种酶来寻找赋予表达的顺式作用序列， 报告基因测定。 因此，可以深入了解控制 其他基因的表达同样局限于“年轻”造血细胞， 细胞 我们将分析STK-1信号转导途径， 地方 我们将研究信号传导的靶点 并希望找到一些新的蛋白质， 好. 最后，我们将测试这个受体参与的假设， 恶性肿瘤或其他涉及造血系统的疾病过程。 STK-1在几种白血病细胞系中表达的发现可能 这表明它在某种程度上参与其中。 RNA和DNA样本来自 患有这些疾病的患者将被测试过表达或 改变受体的表达。