Mechanism of action of TOP2-directed anticancer drugs

TOP2靶向抗癌药物的作用机制

• 批准号: 7006957
• 负责人: LEROY F LIU
• 金额: $ 28.02万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-10 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7006957
• 关键词: DNA damage; DNA topoisomerases; antineoplastics; biological signal transduction; carcinogenesis; cell death; genetically modified animals; laboratory mouse; neoplasm /cancer pharmacology; peptidases; pharmacokinetics; ubiquitin

DESCRIPTION (provided by applicant): The long-term goal of this application is to determine the mechanism of action of topoisomerase II (TOP2)-directed anticancer drugs. In addition to their antitumor activities, TOP2-directed anticancer drugs;uch as VP-16 have been associated with the development of therapy-related acute myelogenous leukaemia (AML). Indeed, our preliminary studies have shown that VP-16 behaves as a stage I tumor promoter in the mouse skin carcinogenesis model. TOP2-directed anticancer drugs are known to trap both TOP2 isozymes, TOP2 alpha and TOP2 beta, into their respective covalent complexes with DNA known as cleavable or cleavage complexes. These two TOP2 isozymes are regulated differently during the cell cycle and most likely perform different functions. However, the differential roles of these two TOP2 isozymes in the antitumor and carcinogenic activities of TOP-directed anticancer drugs remain unclear. Our recent studies have demonstrated that TOP2 beta but not TOP2 alpha cleavage complexes induced by the TOP2-directed anti-cancer drug, VP-16, trigger ubiquitin/26S proteasome-dependent degradation of TOP2 beta (TOP2beta down-regulation). This is the first demonstration of a TOP2 isozyme-specific cellular response to TOP2-directed anticancer drugs. TOP2beta down-regulation is dependent on RNA transcription but not new protein synthesis. Concurrent with TOP2 beta down-regulation, the large subunit of RNA polymerase II, but not other proteins, is also degraded by 26S proteasome. These and other results have suggested that a ubiquitin/26S proteasome pathway(s) is activated locally in the vicinity of the arrested RNA polymerase elongation complexes, leading to the degradation of the TOP2 beta-DNA covalent complex. We further hypothesize that degradation of the TOP2 beta-DNA covalent complex results in exposure of theTOP2-concealed double-strand breaks, which could contribute to both tumor cell death and DNA sequence rearrangements/carcinogenesis. Elucidation of the differential cellular responses to TOP2 cleavage complexes formed by TOP2 isozymes may form the foundation for future development of TOP2 isozyme-specific anticancer drugs. The objective of the current application is to characterize the ubiquitin/26S proteasome pathway induced by TOP2-directed anticancer drugs and to explore the biological consequences of TOP2 beta down-regulation in tumor cell death and carcinogenesis.

描述(申请人提供)：本申请的长期目标是确定拓扑异构酶II(TOP2)导向的抗癌药物的作用机制。除了抗肿瘤活性外，TOP2导向的抗癌药物，如VP-16，也与治疗相关的急性髓系白血病(AML)的发生有关。事实上，我们的初步研究表明，在小鼠皮肤癌变模型中，VP-16作为I阶段肿瘤促进剂发挥作用。众所周知，TOP2导向的抗癌药物可以将TOP2同工酶TOP2α和TOP2β捕获到它们各自的与DNA的共价复合体中，称为可切割或切割复合体。这两种TOP2同工酶在细胞周期中受到不同的调节，很可能发挥不同的功能。然而，这两种TOP2同工酶在顶向抗癌药物的抗肿瘤和致癌活性中的不同作用尚不清楚。我们最近的研究表明，由TOP2导向的抗癌药物VP-16诱导的TOP2β而不是TOP2α裂解复合体可以触发泛素/26S蛋白酶体依赖的TOP2β降解(TOP2beta下调)。这是首次证明TOP2同工酶对TOP2导向的抗癌药物有特异性的细胞反应。TOP2beta的下调依赖于RNA转录，而不是新的蛋白质合成。在TOP2β下调的同时，RNA聚合酶II的大亚基，而不是其他蛋白质，也被26S蛋白酶体降解。这些和其他结果表明，泛素/26S蛋白酶体途径(S)在受阻的RNA聚合酶延长复合体附近被局部激活，导致TOP2β-DNA共价复合体的降解。我们进一步假设，TOP2β-DNA共价复合体的降解导致TOP2隐藏的双链断裂暴露，这可能导致肿瘤细胞死亡和DNA序列重排/致癌。阐明细胞对TOP2同工酶形成的TOP2裂解复合体的不同反应可能为未来TOP2同工酶特异性抗癌药物的开发奠定基础。目前应用的目的是研究TOP2导向的抗癌药物诱导的泛素/26S蛋白酶体途径，并探讨TOP2β下调在肿瘤细胞死亡和癌变中的生物学意义。