NEGATIVE REGULATION OF THE YEAST MRP, YCF1P, AND THE HUMAN MRPS BY CK2

CK2 对酵母 MRP、YCF1P 和人类 MRPS 的负调节

• 批准号: 8360576
• 负责人: Christian M Paumi
• 金额: $ 24.99万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360576
• 关键词: ABCC1 gene; Acute Lymphocytic Leukemia; Biological Models; Cells; Centers of Research Excellence; Clinic; Collaborations; Cyclophosphamide; Dexamethasone; Disease remission; Doxorubicin; Funding; Gleevec; Grant; Human; Imatinib; Inhibitory Concentration 50; Kentucky; Mediating; Molecular; National Center for Research Resources; P-Glycoprotein; Patients; Philadelphia Chromosome; Phosphotransferases; Principal Investigator; Regulation; Research; Research Infrastructure; Resources; Role; Source; Survival Rate; Testing; Treatment Efficacy; United States National Institutes of Health; Universities; Vincristine; Yeasts; base; casein kinase II; chromosome mutation; cost; high risk; human disease; inhibitor/antagonist; outcome forecast; src-Family Kinases; tissue culture

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Treatment of high risk acute lymphoblastic leukemia patients whom are positive for the Philadelphia chromosome mutation (ALL Ph+) which results in the fusion of BCR and Abl kinase (BCR/Abl), patients over the years have been extremely difficult. Recently, the standard hyperCVAD (Cyclophosphamide, Vincristine, Dexamethasone, and Doxorubicin) treatment for these patients was modified to include Gleevec (imatinib). The addition of Imatinib has been extremely successful and has increased the complete remission (CR) rate from 60 to 96% and the 2 year survival rate from about 40% of 85%. The mechanism by which Imatinib increases the efficacy of treatment is unclear. Cyclophosphamide metabolites, doxorubicin, and vincristine are all substrates of multidrug resistance protein 1 (MRP1/ABCC1). Recent studies have shown that increased expression and function of MRP1 is associated with a much poorer prognosis, a reduced CR, and a decreased 2 year survival (over 65% reduction) in ALL patients. Preliminary studies carried out in our lab suggest that BCR/Abl regulates MRP1 function via casein kinase 2 (CK2). CK2 is regulated by BCR/Abl and Src kinases. Interestingly, treatment of ALL Ph+ cells with CK2 inhibitors in combination with Imatinib has been shown to increase Gleevec efficacy (a decrease in the IC50 by 35%) (21). Therefore we hypothesize that inhibition of BCR-Abl with Imatinib down regulates CK2 activity and CK2-mediated induction of MRP1 function, this results in increased cellular accumulation of the CVAD chemotherapeutics (BCR-Abl¿CK2¿MRP1). In this proposal we will: 1) Determine the direct mechanism by which BCR/Abl induces MRP1 function, 2) Determine the role of CK2 in BCR/Abl-mediated induction of MRP1 function, and 3) Test our hypothesis in a Ph+ ALL tissue culture model system, Sup-B15 cells and directly in patient primary cells obtained from the University of Kentucky clinic in collaboration with Dr. Dianna Howard.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 费城染色体突变（ALL Ph+）阳性导致BCR和Abl激酶融合（BCR/Abl）的高危急性淋巴细胞白血病患者的治疗多年来一直极其困难。 最近，针对这些患者的标准 hyperCVAD（环磷酰胺、长春新碱、地塞米松和多柔比星）治疗被修改为包括格列卫（伊马替尼）。 添加伊马替尼非常成功，将完全缓解 (CR) 率从 60% 提高到 96%，2 年生存率从 40% 左右提高到 85%。 伊马替尼提高治疗效果的机制尚不清楚。 环磷酰胺代谢物、阿霉素和长春新碱都是多药耐药蛋白 1 (MRP1/ABCC1) 的底物。 最近的研究表明，MRP1 表达和功能的增加与 ALL 患者的预后较差、CR 降低和 2 年生存率降低（降低超过 65%）相关。 我们实验室进行的初步研究表明，BCR/Abl 通过酪蛋白激酶 2 (CK2) 调节 MRP1 功能。 CK2 受 BCR/Abl 和 Src 激酶调节。 有趣的是，用 CK2 抑制剂与伊马替尼联合治疗 ALL Ph+ 细胞已被证明可以提高格列卫疗效（IC50 降低 35%）(21)。 因此，我们假设用伊马替尼抑制 BCR-Abl 会下调 CK2 活性和 CK2 介导的 MRP1 功能诱导，这导致 CVAD 化疗药物 (BCR-Abl¿CK2¿MRP1) 的细胞积累增加。 在本提案中，我们将：1) 确定 BCR/Abl 诱导 MRP1 功能的直接机制，2) 确定 CK2 在 BCR/Abl 介导的 MRP1 功能诱导中的作用，以及 3) 在 Ph+ ALL 组织培养模型系统、Sup-B15 细胞以及直接在与 Dianna Howard 博士合作从肯塔基大学诊所获得的患者原代细胞中测试我们的假设。