A High Throughput Screen for Telomerase Assembly (RMI)

端粒酶组装 (RMI) 的高通量筛选

• 批准号: 7021596
• 负责人: MICHAEL B JARSTFER
• 金额: $ 7.3万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7021596
• 关键词: DNA binding protein; antineoplastics; biotechnology; drug discovery /isolation; drug screening /evaluation; enzyme activity; enzyme complex; enzyme inhibitors; high throughput technology; molecular assembly /self assembly; neoplasm /cancer chemotherapy; small molecule; technology /technique development; telomerase

DESCRIPTION (provided by applicant): Telomerase is a ribonucleoprotein complex that is specialized for the synthesis of the DNA found on the 3 strand at the end of each chromosome. Telomerase activity is elevated in most cancerous cells while it is absent in most undiseased tissues. Furthermore, inhibiting telomerase activity inhibits growth of cultured cancer cells and prevents growth of human tumors xenografted in mouse models. Thus, telomerase inhibitors have the potential of universal applicability as anticancer therapeutics, and the potential of telomerase inhibition as an anticancer approach has been validated by several approaches. Recently, additional roles for telomerase in promoting tumorigenicity and inhibiting apoptosis have been identified. These roles are independent of DNA synthesis, suggesting that inhibitors of only telomerase enzymatic activity will not achieve the maximum potential effect of telomerase-targeted therapeutics. In this proposal, a new technology platform for inhibiting telomerase activity will be described. The central hypothesis for the experiments is that the in situ generation of a dominant negative telomerase complex will provide a mechanism to disrupt all telomerase-dependent cellular functions. We have developed an assay to follow assemblage of telomerase by observing the association of specific parts of the telomerase complex that are required for telomerase activity. Specific Aim 1 describes efforts to generate a high throughput screen using scintillation proximity assay to report on these essential macromolecular interaction. Specific Aim 2 outlines a secondary screen to further evaluate hits. The completion of these studies will provide a novel HTS that will allow compounds that specific block proper assemblage of the telomerase holoenzyme complex to be identified. Such compounds will serve as leads for anticancer drug discovery and biological studies that illuminate the physiological role of telomerase in mediating tumorigenicity.

描述（由申请人提供）：端粒酶是一种核糖核蛋白复合物，专门用于合成每条染色体末端3链上的DNA。端粒酶活性在大多数癌细胞中升高，而在大多数未患病组织中不存在。此外，抑制端粒酶活性可抑制培养的癌细胞的生长，并防止小鼠模型中异种移植的人肿瘤的生长。因此，端粒酶抑制剂具有作为抗癌治疗剂的普遍适用性的潜力，并且端粒酶抑制作为抗癌方法的潜力已经通过几种方法得到验证。最近，端粒酶在促进致瘤性和抑制细胞凋亡中的其他作用已被确定。这些作用独立于DNA合成，表明仅端粒酶酶活性的抑制剂不会达到端粒酶靶向治疗的最大潜在效果。在这个提议中，将描述用于抑制端粒酶活性的新技术平台。实验的中心假设是显性负性端粒酶复合物的原位产生将提供破坏所有端粒酶依赖性细胞功能的机制。我们开发了一种通过观察端粒酶活性所需的端粒酶复合物的特定部分的关联来跟踪端粒酶组装的测定。具体目标1描述了使用闪烁邻近测定法产生高通量筛选以报告这些基本大分子相互作用的努力。具体目标2概述了一个二级屏幕，以进一步评估命中。这些研究的完成将提供一种新的HTS，这将允许鉴定特异性阻断端粒酶全酶复合物的正确组装的化合物。这些化合物将作为抗癌药物发现和生物学研究的线索，阐明端粒酶在介导致瘤性中的生理作用。