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，结果是触发细胞周期停滞或细胞凋亡，称为检查点反应。我们已经开发了一种基于双杂交系统的酵母试验，它可以检测在某些DNA损伤治疗后某些检查点蛋白之间的相互作用增加。这种现象伴随着检查点的激活，并且可以通过在特殊构建的酿酒酵母菌株的选择性培养基上的菌落生长来检测到。有效的癌症化疗往往依赖于多种药物的组合。这项应用解决了识别具有抗癌细胞活性的小分子以及表征放大或修改其效果的新型试剂的持续需求。我们计划进一步完善大规模机器人筛选的检测条件，并作为一项原则验证研究，解决一个复杂的筛选目标：分离能够增强(或减弱)已有化疗药物喜树碱的检查点激活作用的新化合物。预计这类药物类拓扑异构酶I抑制剂的有效性可以通过提供佐剂来增强其对肿瘤细胞的作用或减弱其对正常细胞的毒性。为此，将追求以下具体目标：1.证明该检测方法发展成为高通量筛选工具的可行性，特别强调筛选能改变喜树碱作用的药物。2.修改和测试适当的仪器，以演示高通量能力。3.筛选化合物文库，寻找能改变喜树碱作用的药物。我们计划在已经向我们提供的第一阶段中筛选国家癌症研究所的多样性化合物集(1900种药物)。随着这个项目的成功完成，我们将建立并验证一个独特的、强大的、具有成本效益的快速筛查系统，并将在分离可开发为喜树碱癌症化疗增强剂或辅助剂的先导化合物方面取得实质性进展。有效的癌症化疗往往依赖于多种药物的组合。这项应用解决了识别具有抗癌细胞活性的小分子以及表征放大或修改其效果的新型试剂的持续需求。北得克萨斯大学健康科学中心(UNTHSC)开发了一项创新技术，可以检测癌症治疗中使用的化疗药物造成的DNA损伤。在这一STTR项目期间，UNTHSC将与当地一家高科技公司(UHV Technologies，Inc.)合作。为了进一步发展这项技术，进行大规模的机器人筛选，以分离能够增强(或减弱)已有化疗药物喜树碱效果的新化合物。UHV将修改其在之前NIH资助下开发的高通量癌症药物发现系统，以通过筛选化合物库来实施和演示这项新技术，以修改喜树碱的效果。