Structural Basis of Multidrug resistance in Cancer

癌症多药耐药性的结构基础

• 批准号: 7939133
• 负责人: OLUWATOYIN Ajibola ASOJO
• 金额: $ 5.4万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939133
• 关键词: ABCG2 gene; ATP phosphohydrolase; ATP-Binding Cassette Transporters; Accounting; Amino Acids; Arginine; Binding; Biological Assay; Breast; Cells; Colon; Complex; Data; Development; Drug Transport; Drug resistance; Environment; Family; Fibroblasts; Genes; Glycine; Glycoproteins; Goals; Human; In Vitro; Institutes; Length; Lung; Malignant Neoplasms; Measures; Membrane; Membrane Transport Proteins; Mentored Research Scientist Development Award; Mentors; Mitoxantrone; Multi-Drug Resistance; Multiple Myeloma; N-terminal; Normal Cell; Nucleotides; Ovarian Carcinoma; Ovary; Pharmaceutical Preparations; Phase; Placenta; Play; Point Mutation; Positioning Attribute; Property; Proteins; Relapse; Research Proposals; Resistance; Roentgen Rays; Role; Sequence Homology; Series; Structure; Testing; Therapeutic; Threonine; Time; anticancer research; base; cancer cell; chemotherapy; cytotoxicity; fibrosarcoma; human ABCG2 protein; in vivo; inhibitor/antagonist; member; mutant; novel; skills; success; three dimensional structure

DESCRIPTION (provided by applicant): Multidrug resistance is a major obstacle to curing cancer because cancer cells become resistant to diverse and unrelated therapeutic compounds. A mechanism for multidrug resistance is the active extrusion of chemotherapeutic drugs from cancer cells by ABC transporters. ABCG2 is a promiscuous ABC transporter of many unrelated compounds. Mutant forms of ABCG2 are expressed in elevated levels in multidrug resistant cancers of diverse origins including fibroblasts, breast, colon, lung, and ovaries. The mechanisms of drug transport remain unclear and the functions of each domain of ABCG2 are neither tested nor confirmed. Furthermore, there are no 3-dimensional structures of ABCG2. The following specific aims are proposed to rectify this situation: 1) To characterize full length ABCG2 and its domains. The activity of each domain of ABCG2 will be tested using assays that measure cytotoxicity, ATPase activity, drug binding and drug extrusion. 2) To determine 3-dimensional structures of ABCG2's cytosolic domain. X-ray structures will be solved that reveal the mode of binding of nucleotides to the cytosolic domain. 3) To determine 3-dimensional structures of full length ABCG2 and domains. Structures will be solved of full length and active domains of ABCG2. The correlation of structural and activity data will clarify the mechanism of drug transport by ABCG2 and homologous transporters, thus spearheading new strategies for the development of novel chemotherapies for multidrug resistant cancer. Specific aims 1 and 2 are proposed for Phase I, while specific aim 3 is proposed for Phase II of the K01 award and beyond. The K01 will afford the applicant protected time to develop new skills and to apply existing skills to cancer research, with the guidance of her mentors, at the stimulating educational environment of the Eppley Institute. She will then be ready to successfully compete for and obtain an independent tenure track position in cancer research.

描述（由申请人提供）：多药耐药性是治愈癌症的主要障碍，因为癌细胞对多种和无关的治疗化合物具有抗性。多药耐药性的机制是ABC转运蛋白从癌细胞中积极挤出化学治疗药物。 ABCG2是许多无关化合物的ABC转运蛋白。 ABCG2的突变形式在多种起源的多种抗药性癌中以较高的水平表达，包括成纤维细胞，乳房，结肠，肺和卵巢。药物转运的机制尚不清楚，并且既未测试也没有证实ABCG2的每个结构域的功能。此外，没有ABCG2的三维结构。提出了以下特定目标来纠正这种情况：1）表征全长ABCG2及其域。 ABCG2每个结构域的活性将使用测量细胞毒性，ATPase活性，药物结合和药物挤出的测定法进行测试。 2）确定ABCG2胞质结构域的3维结构。将解决X射线结构，以揭示核苷酸与胞质结构域结合的方式。 3）确定全长ABCG2和域的3维结构。结构将由ABCG2的全长和活跃域求解。结构和活性数据的相关性将阐明ABCG2和同源转运蛋白的药物转运机制，从而为开发用于多药耐药性癌症的新型化学疗法的新策略。提出了针对第一阶段的特定目的1和2，而针对K01奖的II阶段则提出了特定的目标3。 K01将为申请人提供培养新技能并在埃普利研究所（Eppley Institute）刺激的教育环境中的指导下，在癌症研究中运用新技能并将现有技能应用于癌症研究。然后，她将准备成功竞争并获得癌症研究中的独立终身轨道位置。

项目成果

Expression, purification, crystallization and preliminary X-ray analysis of a truncated soluble domain of human glioma pathogenesis-related protein 1.

人胶质瘤发病机制相关蛋白1截短可溶结构域的表达、纯化、结晶和初步X射线分析。

• DOI: 10.1107/s1744309110035669
• 发表时间: 2010
• 期刊: Acta crystallographica. Section F, Structural biology and crystallization communications
• 作者: Bonafe,Nathalie;Zhan,Bin;Bottazzi,MariaElena;Perez,OrianaA;Koski,RaymondA;Asojo,OluwatoyinA
• 通讯作者: Asojo,OluwatoyinA