Modeling the Responsiveness of Sensitive Populations to Genotoxic Agents Using DNA Repair Inhibitors

使用 DNA 修复抑制剂模拟敏感人群对基因毒性药物的反应性

基本信息

批准号：
10734425
负责人：
Jeffrey C Bemis
金额：
$ 23.03万
依托单位：
LITRON LABORATORIES, LTD.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10734425
关键词：
Address Alkylating Agents Alkylation Antimetabolites Apoptosis Area Base Excision Repairs Benchmarking Biological Assay Biological Markers Cell Line Cell Nucleus Cells Chemical Exposure Chemicals Computer software Crosslinker DNA DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway Dose Drug or Chemical Engineering Exhibits Family Fee-for-Service Plans Future Genetic Genetic Variation Genomics Goals Histone H3 Hour Human Human Cell Line In Vitro Individual Differences Laboratories Mammalian Cell Medicine Metaphase Methods Modeling Mutagenicity Tests Mutagens Nonhomologous DNA End Joining Oxidants Oxidative Stress Pathway interactions Poison Population Predisposition Reagent Rodent Rodent Model Scheme Source Specificity Stress TP53 gene Testing Time Topoisomerase II Toxic effect Toxicogenetics Toxicology Type I DNA Topoisomerases United States National Academy of Sciences Work cost effective cytotoxicity data analysis pipeline experimental study genotoxicity inhibitor innovation novel pre-clinical preservation repaired response small molecule testing services

项目摘要

Project Summary It is well recognized that conventional toxicology assays exhibit a crucial deficiency. Specifically, their use of limited numbers of cell lines and rodent strains does not provide information about toxic responses that may occur in sensitive human populations. Although studying very large numbers of cell lines and laboratory rodent models could theoretically address this issue, it is not a practical answer. There are simply too many legacy chemicals and new chemical substances being developed for this to represent a routine testing scheme. Fortunately, in the area of genetic toxicology, a solution exists. Our innovative approach takes into account the fact that inter-subject differences in sensitivity to genotoxic agents is predominately due to variable DNA repair capacity. Our project will develop an efficient, high throughput, in vitro genotoxicity assay platform that provides information about chemicals’ potential to cause DNA damage in prototypical and sensitive human populations. The high efficiency of our proposed approach is in part explained by our decision to study a single p53- competent human cell line (TK6) to generate prototypical genotoxic response profiles. Sensitive population responses will be modelled by co-exposing cells to each test chemical in combination with a panel of small molecule DNA repair inhibitors. These inhibitors will be chosen to cover each of the major DNA repair pathways. In addition to our use of DNA repair inhibitors to efficiently mimic sensitive populations, other innovative aspects of our project include our use of a multiplexed DNA damage assay to cover multiple biomarkers of genotoxicity and cytotoxicity, and state-of-the-art benchmark dose software to characterize the family of dose-response relationships that will be generated for each chemical studied. Once reduced to practice, we will make the assay available as a set of kit-formatted reagents, and also through fee-for-service testing.

项目摘要众所周知，常规毒理学测定法表现出至关重要的缺陷。具体来说，他们的使用有限数量的细胞系和啮齿动物菌株不会提供有关可能有毒反应的信息发生在敏感人群中。尽管研究了大量的细胞系和实验室啮齿动物从理论上讲，模型可以解决这个问题，这不是一个实际的答案。有太多遗产为此开发了化学物质和新的化学物质，以代表常规测试方案。幸运的是，在遗传毒理学领域，存在溶液。我们的创新方法考虑了事实认为，对遗传毒性剂的敏感性差异主要是由于可变的DNA修复容量。我们的项目将开发一个高效，高通量的体外遗传毒性测定平台，以提供有关化学药品在原型和敏感人群中引起DNA损伤的潜力的信息。我们提出的方法的高效率在某种程度上是通过我们研究单个p53-的决定来解释的。主管人细胞系（TK6）生成典型的遗传毒性反应谱。敏感人群响应将通过共暴露细胞对每种测试化学化学物质的共同模型，并结合一组小面板分子DNA修复抑制剂。这些抑制剂将被选择覆盖每个主要的DNA修复途径。除了我们使用DNA修复抑制剂有效地模仿敏感人群，其他我们项目的创新方面包括使用多路复用DNA损伤测定法来覆盖多个遗传毒性和细胞毒性的生物标志物以及最先进的基准剂量软件以表征每个化学研究生将产生的剂量反应关系家族。一旦降低到练习，我们将使Assame作为一组套件形式的试剂，也可以通过支付费用提供测试。