Rapid Screen for Genotoxicants, Chemoprotectors, and Radioprotectors

描述（由申请人提供）：该研究项目解决了化学和制药行业面临的一个重要问题-他们需要在较短的时间内评估不断增加的新化学实体的DNA损伤潜力。这种情况为能够为这些挑战提供解决方案的小企业提供了机会。我们的建议回答了工业界的需要，通过开发一个自动化的体外染色体损伤检测的高通量毒性评估。将通过流式细胞仪微核测量定量染色体损伤。重要的是，将在本项目过程中开发的方法不仅代表了高容量遗传毒性筛选系统，而且还可以配置为识别防止染色体损伤的试剂，即生物反应调节剂（BRM）。BRM提供了化疗或放射防护的好处，在几个重要领域有很大的希望，包括治疗癌症患者，在战争期间保护我们的部队，以及治疗化学或放射性恐怖袭击的平民受害者。拟议的第二阶段实验最初将侧重于开发方法，使我们能够将自动液体处理设备纳入小型化检测。通过消除转移步骤和其他劳动密集型任务，该分析将满足具有高容量测试要求的制药和化学公司的规格。一旦确定，我们和我们在制药行业的合作者将评估一组遗传毒性物质和非遗传毒性物质，从而提供有关检测便携性，灵敏度和特异性的信息。此外，还将进行另外两项研究：评价HepG 2肝细胞系;以及利用微核评分试验进行辐射防护剂鉴别/表征。最终，这种筛选方法的优化和验证将有助于化学和制药公司将其资源分配给最有前途的候选人，从而在开发早期消除危险实体。此外，通过帮助识别下一代BRM，癌症患者和武装部队将受益于化疗和放射保护药物的可用性。沿着分子生物学、化学和医学领域的最新进展，出现了越来越多的新的和潜在有用的药物和工业化学品。制药和化学公司在安全性和有效性方面优先考虑这些制剂的能力往往因使用过时的方法和/或技术而受到阻碍。该提案解决了自动化和小型化体外微核试验的需求，旨在快速有效地检测药物和化学品的染色体损伤潜力。此外，该测定可用于研究旨在改善辐射暴露影响的试剂，例如生物反应调节剂。

DESCRIPTION (provided by applicant): This research project addresses an important problem faced by the chemical and drug industries-their requirement to evaluate the DNA damaging potential of ever increasing numbers of new chemical entities in shorter periods of time. This situation presents opportunities for small businesses that are able to provide solutions to these challenges. Our proposal answers industry's need for higher throughput toxicity assessment through the development of an automated in vitro chromosomal damage assay. Chromosome damage will be quantified by flow cytometric micronucleus measurements. Importantly, the methodology that will be developed over the course of this project not only represents a high volume genotoxicant screening system, but also one that can be configured to identify agents which protect against chromosomal damage, i.e. biological response modifiers (BRMs). BRMs provide chemo- or radio-protection benefits that hold great promise in several important arenas, including treatment of cancer patients, protection of our troops during times of war, and for treatment of civilian victims of chemical or radiological terrorist attacks. The proposed Phase II experiments will initially focus on developing the methodologies that will allow us to incorporate automatic liquid handling devices into a miniaturized assay. By eliminating transfer steps and other labor-intensive tasks, the assay will meet the specifications of pharmaceutical and chemical companies that have high volume testing requirements. Once this is established, we, and our collaborators in the pharmaceutical industry, will evaluate a panel of genotoxicants and non-genotoxicants, thereby providing information regarding assay portability, sensitivity, and specificity. Additionally, two other lines of investigation will be undertaken: evaluation of the HepG2 liver cell line; and utilization of the micronucleus scoring assay for the purposes of radioprotectant identification/characterization. Ultimately, optimization and validation of this screening assay will help chemical and drug companies allocate their resources to their most promising candidates, eliminating hazardous entities early in development. Furthermore, by aiding in the identification of next generation BRMs, cancer patients and armed forces will benefit from the availability of chemo- and radio-protectant drugs. Along with recent advancements in the fields of molecular biology, chemistry and medicine comes an ever- increasing number of new and potentially helpful drugs and industrial chemicals. The ability of pharmaceutical and chemical companies to prioritize these agents in terms of safety and efficacy is often hampered by the use of outdated methods and/or technologies. This proposal addresses the stated need for an automated and miniaturized in vitro micronucleus assay designed to rapidly and efficiently detect the chromosome damaging potential of drugs and chemicals. In addition, this assay can be used to investigate agents designed to ameliorate the effects of radiation exposure, e.g. biological response modifiers.