Inhibition of Recombination DNA Repair in Pancreatic Cancer Cells

抑制胰腺癌细胞中的重组 DNA 修复

• 批准号: 7315702
• 负责人: Philip P Connell
• 金额: $ 12.28万
• 依托单位: UNIVERSITY OF CHICAGO
• 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 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7315702
• 关键词: Biological Assay; Cancer Etiology; Cells; Cessation of life; Clinical Trials; DNA Damage; DNA Repair; Data; Data Analyses; Development; Filament; Goals; Grant; Human; In Vitro; Inhibitory Concentration 50; Libraries; Liquid substance; Malignant neoplasm of pancreas; Outcome; Patients; Pharmaceutical Preparations; Progress Reports; Proteins; Rad51 recombinase; Radiation therapy; Research; Resistance; Robotics; Screening procedure; Single-Stranded DNA; Specificity; Standards of Weights and Measures; Techniques; Testing; United States; Work; base; cancer cell; chemotherapy; high throughput screening; improved; inhibitor/antagonist; monomer; oncology; pancreatic neoplasm; recombinational repair; small molecule; small molecule libraries; therapy resistant; tumor

DESCRIPTION (provided by applicant): Pancreatic cancer remains the 4th leading cause of cancer death in the United States. These tumors appear to be relatively resistant to standard available therapies. An agent capable of sensitizing pancreatic cancer cells to oncology therapies may improve the outcomes of these patients. A central hypothesis of this project is that elevated levels of homologous recombinational (HR) DNA repair can cause human pancreatic tumors to be resistant to some chemotherapies and radiotherapy, and that specific inhibition of HR may help overcome this resistance to therapy. The proposed research plan will develop a multi-step screen for identifying small molecule inhibitors of human RAD51, which is the central protein involved in initiating HR. The initial submission of this project included an aim proposing to develop a high-throughput (HT) assay and to screen a na¿ve library of small molecules in search of compounds that can inhibit the formation of RAD51 filaments on single-stranded DNA. A portion of initial work has been completed and is now summarized in the Preliminary Studies / Progress Report section. The screen of a 10k library identified 72 compounds that inhibit RAD51 filament formation by = 50%. This work successfully validated the filament formation assay as an HT screen, and it provided important information including refinement of assay techniques and data analysis. Using this information, the screen will be now be repeated using a larger starting library (130k compounds) and robotic liquid handling, which will likely yield compounds with even greater activities and specificities (Aim 1). To determine which have specific activities, the compounds resulting from both screens will be tested further in secondary and tertiary assays. Aim 2 will employ a set of in-vitro secondary assays aimed at identifying those compounds that can specifically block particular functions of purified RAD51 protein. In the third aim, compounds will be further characterized with tertiary cell-based assays, to identify the subset capable of sensitizing pancreatic cancer cells to the lethal effects of DNA damaging therapies via HR-specific mechanisms. The ultimate goal is to identify at least one optimal compound suitable for further development in clinical trials as an oncology drug. Pancreatic cancer remains the 4th leading cause of cancer death in the United States. These tumors appear to be relatively resistant to presently available therapies. A drug capable of overcoming the cellular resistance to radiotherapy and/or chemotherapeutic drugs may improve on the outcomes of these patients. The goal of this proposal is to develop an agent capable of sensitizing pancreatic cancer cells to these common oncology therapies.

描述（由适用提供）：胰腺癌仍然是美国癌症死亡的第四个主要原因。这些肿瘤似乎对标准可用疗法具有相对抗性。能够使胰腺癌细胞对肿瘤疗法敏感的药物可以改善这些患者的结果。该项目的一个核心假设是，同源重组（HR）DNA修复水平升高会导致人胰腺肿瘤对某些化学疗法和放射疗法具有抗性，并且对人力资源的特定抑制可能有助于克服这种对治疗的抵抗力。拟议的研究计划将开发一个多步筛选，用于鉴定人RAD51的小分子抑制剂，该筛选是涉及启动HR的中心蛋白。该项目的最初提交包括一项针对开发高通量（HT）测定法的目标建议，并筛选小分子的Na ve库，以寻找可以抑制单链DNA上Rad51细丝形成的化合物。初始工作的一部分已经完成，现在在初步研究 /进度报告部分中进行了总结。 10K库的屏幕确定了72种抑制RAD51细丝形成的化合物= 50％。这项工作成功验证了细丝形成分析作为HT屏幕，并提供了重要信息，包括对测定技术和数据分析的细化。使用此信息，现在将使用较大的起始库（130K化合物）和机器人液体处理重复屏幕，这可能会产生具有更大活动和特异性的化合物（AIM 1）。为了确定哪些具有特定的活动，将在第二和第三纪测定中进一步测试两个筛选的化合物。 AIM 2将采用一组体外次级测定，旨在识别可以特异性阻断纯化RAD51蛋白的特定功能的化合物。在第三个目的中，将通过基于三级细胞的测定进一步表征化合物，以鉴定能够通过HR特异性机制将胰腺癌细胞敏化DNA损伤疗法的致命作用的子集。最终目标是将至少一种适合在临床试验中进一步开发的最佳化合物作为肿瘤药物。胰腺癌仍然是美国癌症死亡的第四个主要原因。这些肿瘤似乎对目前可用的疗法具有相对抗性。能够克服对放射疗法和/或化学治疗药物的细胞耐药性的药物可能会改善这些患者的结果。该提案的目的是开发能够将胰腺癌细胞敏化的药物对这些常见的肿瘤疗法。