A Two Hybrid System Based Yeasy Screen for Agents that Affect DNA Damage Checkpoi

基于两种混合系统的 Yeasy 筛选影响 DNA 损伤检查点的试剂

• 批准号: 7326735
• 负责人: NALIN KUMAR
• 金额: $ 13.54万
• 依托单位: UHV TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7326735
• 关键词: Address; Adjuvant; Affect; Agar; Amplifiers; Antineoplastic Agents; Apoptosis; Automation; Biological Assay; Businesses; Camptothecin; Cancer Control; Cell Cycle Arrest; Cells; Chemotherapy-Oncologic Procedure; Class; Condition; DNA Binding Domain; DNA Damage; Development; Family; Funding; Future; Goals; Growth; Health Sciences; Histidine; Image Analysis; Lead; Libraries; Malignant Neoplasms; Molecular; National Cancer Institute; Normal Cell; Numbers; Pharmaceutical Preparations; Phase; Plasmids; Proteins; Radiation therapy; Reporter; Robotics; Saccharomyces cerevisiae; Sales; Screening procedure; Services; Small Business Technology Transfer Research; Source; System; Techniques; Technology; Testing; Texas; Therapeutic Index; Topoisomerase-I Inhibitor; Toxic effect; United States National Institutes of Health; Universities; Work; Yeasts; base; cancer cell; cancer therapy; chemotherapeutic agent; chemotherapy; cost efficient; drug discovery; high throughput screening; improved; inhibitor/antagonist; innovation; instrument; instrumentation; interest; neoplastic cell; novel; response; small molecule; tool; yeast two hybrid system

DESCRIPTION (provided by applicant): Cancer chemotherapy agents frequently introduce DNA damage and, as a consequence, trigger cell cycle arrest or apoptosis, referred to as checkpoint responses. We have developed a two-hybrid-system based yeast assay that detects increased interaction between certain checkpoint proteins following certain DNA-damaging treatments. This phenomenon accompanies checkpoint activation and can be detected by colony growth on selective medium in specially constructed strains of the yeast Saccharomyces cerevisiae. Effective cancer chemotherapy frequently relies on a combination of agents. This application addresses the continued need to identify small molecules that have activity against cancer cells and to characterize novel agents that amplify or modify their effect. We plan to further refine the assay conditions for large-scale robotic screening and, as a proof-of-principle study, to address a sophisticated screening goal: to isolate novel compounds that enhance (or diminish) the checkpoint-activating effect of the established chemotherapy drug camptothecin. It is expected that the usefulness of this agent class of topoisomerase I inhibitors can be enhanced by providing adjuvants that enhance its effect on tumor cells or blunt its toxicity towards normal cells. To this end, the following specific aims will be pursued: 1. To demonstrate the feasibility of developing the assay into a high- throughput screening tool, with special emphasis on screening for agents that modify the camptothecin effect. 2. To modify and test the appropriate instrumentation for demonstration of high throughput capability. 3. To screen a compound library for agents that modify the effects of camptothecin. We plan to screen the National Cancer Institute's Diversity Compound set (1,900 agents) during Phase I that is already available to us. At the successful completion of this project, we will have established and validated a unique, robust, cost efficient and rapid screening system and will have made substantial progress towards isolation of lead compounds that can be developed into amplifiers or adjuvants in camptothecin cancer chemotherapy. Effective cancer chemotherapy frequently relies on a combination of agents. This application addresses the continued need to identify small molecules that have activity against cancer cells and to characterize novel agents that amplify or modify their effect. University of North Texas Health Science Center (UNTHSC) has developed an innovative technique that detects DNA damage caused by chemotherapy agents used in cancer treatment. During this STTR project, UNTHSC will work with a local high technology company (UHV Technologies, Inc.) to further develop this technique for large-scale robotic screening to isolate novel compounds that enhance (or diminish) the effects of the established chemotherapy drug camptothecin. UHV will modify its high throughput cancer drug discovery system developed under previous NIH funding to implement and demonstrate this new technique by screening a compound library for agents that modify the effects of camptothecin.

描述（由申请人提供）：癌症化疗药物经常引起 DNA 损伤，从而引发细胞周期停滞或细胞凋亡，称为检查点反应。我们开发了一种基于双杂交系统的酵母检测，可检测某些 DNA 损伤处理后某些检查点蛋白之间相互作用的增加。这种现象伴随着检查点激活，并且可以通过在特殊构建的酿酒酵母菌株中的选择性培养基上的菌落生长来检测。有效的癌症化疗通常依赖于多种药物的组合。该申请解决了识别具有抗癌细胞活性的小分子以及表征增强或改变其作用的新型药物的持续需求。我们计划进一步完善大规模机器人筛选的测定条件，并作为原理验证研究，解决复杂的筛选目标：分离出增强（或减弱）已建立的检查点激活作用的新型化合物。化疗药物喜树碱。预期可以通过提供增强其对肿瘤细胞的作用或减弱其对正常细胞的毒性的佐剂来增强此类拓扑异构酶I抑制剂的有用性。为此，将追求以下具体目标： 1. 证明将该测定开发为高通量筛选工具的可行性，特别强调筛选改变喜树碱效应的药物。 2. 修改和测试适当的仪器以演示高通量能力。 3.筛选化合物库以寻找改变喜树碱作用的药物。我们计划在第一阶段筛选我们已经可用的国家癌症研究所的多样性化合物组（1,900 种药物）。该项目成功完成后，我们将建立并验证一个独特、稳健、具有成本效益的快速筛选系统，并将在分离可开发为喜树碱癌症化疗中的放大器或佐剂的先导化合物方面取得实质性进展。有效的癌症化疗通常依赖于多种药物的组合。该申请解决了识别具有抗癌细胞活性的小分子以及表征增强或改变其作用的新型药物的持续需求。北德克萨斯大学健康科学中心 (UNTHSC) 开发了一种创新技术，可以检测癌症治疗中使用的化疗药物引起的 DNA 损伤。在这个 STTR 项目期间，UNTHSC 将与当地一家高科技公司 (UHV Technologies, Inc.) 合作，进一步开发这项技术，用于大规模机器人筛选，以分离出增强（或减弱）现有化疗药物喜树碱效果的新型化合物。 UHV 将修改其在 NIH 先前资助下开发的高通量癌症药物发现系统，通过筛选化合物库来寻找可改变喜树碱作用的药物，以实施和演示这项新技术。