High Throughput Screens of Novel Radiation Sensitizers and Protectors

新型辐射敏化剂和保护剂的高通量筛选

• 批准号: 7911884
• 负责人: William F Morgan
• 金额: $ 29.89万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-05 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7911884
• 关键词: Accidents; Alleles; Arts; Baltimore; Biochemical Pathway; Biological Assay; Cell Line; Cell Survival; Cells; Chromosomal Instability; Clinic; Clinical; Devices; Dose; Emergency Situation; Event; Exposure to; FDA approved; Fluorescence; Genes; Genomic Instability; Goals; Human; Individual; Ionizing radiation; Lead; Libraries; Malignant Neoplasms; Maryland; Measures; Micronucleus Tests; Military Personnel; Molecular Target; Mutation; Normal tissue morphology; Nuclear; Pathway interactions; Patients; Pharmaceutical Preparations; Preclinical Drug Evaluation; Proteins; Public Health; Radiation; Radiation Oncology; Radiation therapy; Radiation-Protective Agents; Radiation-Sensitizing Agents; Radio; Radioactive; Radiobiology; Reporter; Research; Robotics; Technology; Terrorism; Testing; Time; Universities; base; cancer cell; cell killing; cell type; clinically relevant; dirty bomb; drug discovery; emergency service responder; high throughput screening; improved; interest; irradiation; killings; neoplastic cell; novel; novel therapeutics; radiation effect; tumor

DESCRIPTION (provided by applicant): This is a new application that brings together expertise in radiation biology and robotic high throughput drug screen technology to identify new radiation sensitizers and protectors for use in clinical radiation oncology and in the event of radiological terrorism. Three specific aims are proposed. Specific Aim 1 will implement a high throughput cell-based screen to identify compounds that can sensitize/protect human cells to/from ionizing radiation. This is a drug discovery aim that will allow us to identify lead molecules that are radio-sensitizers or radio-protectors from a library consisting of ~40,000 compounds. Specific Aim 2 will validate the results obtained in Aim 1 using clonogenic cell survival assays. Different drug concentrations and exposure times will be investigated at increasing radiation doses. Aim 2 will also test the hypothesis that radiation protectors are effective when used after irradiation. Once we have confirmed the initial observations regarding sensitization or protection, we will use isogenic cells that lack one or both alleles of genes known to be disrupted in tumor cells to determine whether lead compounds sensitize/protect differentially in wild type cells versus cells with well characterized mutations commonly found in cancer. Specifically we will test the hypothesis that compounds will differentially sensitize/protect wild type cells compared with cells containing genetic alterations commonly found in cancer cells. Specific Aim 3 will test the hypothesis that the sensitizers/protectors identified in Aim 1 and confirmed in Aim 2 will not predispose surviving cells to enhanced genomic instability. Exposure to clinically relevant doses of radiation in the presence of sensitizers or protectors will not kill all cells. Using a green fluorescence protein-based reporter assay we will test the hypothesis that the progeny of cells surviving irradiation +/-sensitizer or protector are not more likely to manifest genomic instability as measured by delayed mutation and hyperrecombination. Using a micronucleus assay we will test the hypothesis that the progeny cells surviving irradiation +/- sensitizer or protector are not more likely to manifest genomic instability as measured by delayed chromosomal instability. All technology and cell lines are available and in place at the University of Maryland. We have the expertise to identify new radiation sensitizers/protectors for use in the clinic and in the event of radiological terrorism. Successfully achieving the proposed research goals will be a significant contribution to patients undergoing radiation therapy and to the public in the event of terrorism involving radioactive material. PUBLIC HEALTH RELEVENCE: There are two feasible and potentially very significant goals to this application. The first is to use state of the art technologies to identify compounds that can sensitize cancer cells to radiation- induced cell killing and thus improve radiation therapy. The second is to use those same technologies to find unique compounds that can protect against deleterious radiation effects and thus protect individuals in the event of a radiological incident.

描述（由申请人提供）：这是一项新的应用，汇集了放射生物学和机器人高通量药物筛选技术的专业知识，以识别用于临床放射肿瘤学和放射恐怖主义事件的新型放射增敏剂和保护剂。提出了三个具体目标。具体目标1将实施高通量的基于细胞的筛选，以确定可以使人体细胞对电离辐射敏感/保护人体细胞免受电离辐射的化合物。这是一个药物发现的目标，将使我们能够从由约40，000种化合物组成的库中识别出作为辐射增敏剂或辐射保护剂的先导分子。特定目标2将使用克隆原性细胞存活试验验证目标1中获得的结果。将在增加辐射剂量的情况下研究不同的药物浓度和暴露时间。目标2还将检验辐射防护器在辐照后使用时有效的假设。一旦我们证实了关于致敏或保护的初步观察结果，我们将使用缺乏已知在肿瘤细胞中被破坏的基因的一个或两个等位基因的同基因细胞来确定先导化合物是否在野生型细胞与具有在癌症中常见的充分表征的突变的细胞中差异地致敏/保护。具体而言，我们将测试以下假设：与含有癌细胞中常见的遗传改变的细胞相比，化合物将差异性地敏化/保护野生型细胞。特定目标3将检验目标1中鉴定并在目标2中确认的致敏剂/保护剂不会使存活细胞倾向于增强基因组不稳定性的假设。在存在敏化剂或保护剂的情况下，暴露于临床相关剂量的辐射不会杀死所有细胞。使用基于绿色荧光蛋白的报告基因测定，我们将检验这样的假设，即在辐射+/-敏化剂或保护剂下存活的细胞的后代不太可能表现出通过延迟突变和过度重组测量的基因组不稳定性。使用微核试验，我们将检验以下假设：存活于辐射+/-敏化剂或保护剂的子代细胞不太可能表现出通过延迟染色体不稳定性测量的基因组不稳定性。所有的技术和细胞系都可以在马里兰州大学获得和使用。我们拥有识别新的辐射敏化剂/保护剂的专业知识，可用于临床和放射恐怖主义事件。成功实现拟议的研究目标将对接受放射治疗的病人和在涉及放射性材料的恐怖主义事件中对公众作出重大贡献。公共卫生解放：这一应用有两个可行的和潜在的非常重要的目标。第一个是使用最先进的技术来鉴定可以使癌细胞对辐射诱导的细胞杀伤敏感的化合物，从而改善放射治疗。第二个目标是利用同样的技术来寻找独特的化合物，以防止有害的辐射影响，从而在发生辐射事件时保护个人。