THE ROLE OF PL3K IN THROMBOPOIETIN SIGNALING

PL3K 在血小板生成素信号传导中的作用

• 批准号: 6226398
• 负责人: AMY E GEDDIS
• 金额: $ 12.58万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6226398
• 关键词: apoptosis; biological signal transduction; cell proliferation; density gradient ultracentrifugation; enzyme activity; flow cytometry; gel mobility shift assay; immunoprecipitation; laboratory mouse; mitogen activated protein kinase; northern blottings; p53 gene /protein; phosphatidylinositol 3 kinase; phosphorylation; polymerase chain reaction; protein structure function; thrombopoietic factor; tissue /cell culture; western blottings

(Adapted from applicant's abstract) This K08 proposal outlines a research plan designed to give the applicant, Dr. Amy E. Geddis, the scientific training and career development necessary for a successful career in academic medicine and basic science. The proposed research will characterize the role of phosphatidylinositol 3-kinase (P13K) in cell survival and proliferation in response to thrombopoietin (TPO). TPO is the primary regulator of megakaryo- cyte (MK) development and plays an important role in hematopoiesis in general, but the mechanisms by which its receptor, Mpl, signals to stimulate cell survival and proliferation are incompletely understood. Our preliminary data suggest a role for P13K in these processes. The proposal outlines a strategy combining the use of kinase inhibitors and inducible expressed dominant negative kinases to identify elements of the P13K pathway that are important for its function in mediating survival and proliferation in response to TPO. In addition, several candidate pathways in apoptosis and proliferation will be evaluated for P13K-dependent regulation by TPO. Finally, the platelet lowering drug Anagrelide, will be evaluated for its potential interference with P13K signaling by TPO. Proposed studies will involve both Mpl-expressing cell lines as well as primary murine MK progenitors. The results of these studies will lead to a better understanding of the mechanisms by which TPO signals to promote cell survival and proliferation. Characterization of TPO signaling is critical in order to understand normal MK development, and to understand the implications of therapeutic modulation of thrombopoiesis for example involving the clinical use of TPO or Anagrelide. The University of Washington provides a strong research environment for this proposal with many faculty members engaged in related studies (hematopoiesis, signal transduction, biochemistry, apoptosis, molecular biology) whose advise and guidance will be available. There are ongoing seminars and courses in relevant areas, and the proximity of the Fred Hutchinson Cancer Research Center provides an additional resource. Dr. Kaushansky and the advisory panel, formed specifically for the aims of this proposal, will oversee the applicant's research and academic progress. A defined plan of training is presented in which the applicant will develop the experience and skills necessary for success as an independent investigator in Hematology.

（改编自申请人的摘要）此 K08 提案概述了一项研究计划 旨在为申请人 Amy E. Geddis 博士提供科学培训和 学术医学事业成功所必需的职业发展 基础科学。 拟议的研究将描述以下角色的作用： 磷脂酰肌醇 3-激酶 (P13K) 在细胞存活和增殖中的作用 对血小板生成素（TPO）的反应。 TPO 是巨核细胞的主要调节因子 细胞 (MK) 发育并在一般造血过程中发挥重要作用， 但其受体 Mpl 发出刺激细胞信号的机制 生存和增殖尚不完全清楚。 我们的初步数据 表明 P13K 在这些过程中的作用。 该提案概述了一项战略 结合使用激酶抑制剂和诱导表达显性 阴性激酶来识别重要的 P13K 通路元件 因其在 TPO 反应中介导生存和增殖的功能。 此外，细胞凋亡和增殖的几个候选途径也将被确定。 评估 TPO 的 P13K 依赖性调节。 最后是血小板 降低药物阿那格雷，将评估其潜在干扰 通过 TPO 进行 P13K 信号传导。 拟议的研究将涉及 Mpl 表达 细胞系以及原代鼠 MK 祖细胞。 这些结果 研究将有助于更好地理解 TPO 的机制 促进细胞存活和增殖的信号。 TPO 的表征 为了了解正常的 MK 发育并了解 了解血小板生成治疗调节的影响 例如涉及 TPO 或阿那格雷的临床使用。 大学 华盛顿为该提案提供了强大的研究环境，许多人 从事相关研究（造血、信号 转导、生物化学、细胞凋亡、分子生物学）谁的建议和 将提供指导。 正在进行的研讨会和课程 相关区域以及 Fred Hutchinson 癌症研究中心的邻近性 中心提供额外资源。 考尚斯基博士和顾问 专门为此提案而成立的小组将监督 申请人的研究和学术进展。 明确的培训计划是 申请人将在其中发展经验和技能 作为血液学独立研究者的成功所必需的。