Validation of a Rodent Mutagenicity Assay

啮齿动物致突变性测定的验证

• 批准号: 8070097
• 负责人: STEPHEN D DERTINGER
• 金额: $ 71.91万
• 依托单位: LITRON LABORATORIES, LTD.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8070097
• 关键词: Academia; Address; Affinity; Aging; Anabolism; Animals; Atherosclerosis; Biological Assay; Blood; Cell Line; Cell Lineage; Cell surface; Cells; Chemical Industry; Chemicals; DNA; DNA Damage; Data; Databases; Development; Disease; Dose; Drug Industry; Enrollment; Environment; Erythrocytes; Erythroid; Event; Exposure to; Family suidae; Feedback; Flow Cytometry; Genes; Glycosylphosphatidylinositols; Government; Guidelines; Harvest; Hepatocyte; Human Resources; Industry; Instruction; International; Investigation; Laboratories; Laboratory Study; Life; Ligands; Lymphocyte; Malignant Neoplasms; Measures; Methodology; Methods; Modeling; Mutagenesis; Mutagens; Mutate; Mutation; Nitrosourea Compounds; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Poison; Population; Preparation; Primary carcinoma of the liver cells; Process; Production; Proteins; Protocols documentation; Public Health; Qualifying; Rat Strains; Rattus; Reagent; Reporter; Reproducibility; Research Project Grants; Resistance; Reticulocytes; Rodent; Safety; Sales; Schedule; Scoring Method; Small Business Innovation Research Grant; Societies; Speed; Staging; Staining method; Stains; System; Time; Tissues; Toxicology; Validation; Work; Workplace; X Chromosome; aerolysin; analytical method; base; cell type; cost effective; cost effectiveness; design; genotoxicity; improved; in vivo; interest; killings; mutant; novel; peripheral blood; pre-clinical; preference; prevent; public health relevance; research study; success; symposium; tissue culture; validation studies; willingness

DESCRIPTION (provided by applicant): Mutation to DNA is a primary mechanism by which cancers arise. These events have also been implicated in diseases such as atherosclerosis, and processes such as aging. Therefore, there is an important need for sensitive analytical methods that facilitate the study of mutagenesis, as well as the identification of chemical or physical agents that can mutate DNA. Methods for measuring in vivo mutation currently exist, each with their own advantages and limitations. While some are based on colony formation and require tissue culture work, others rely on expensive, proprietary trangenic rodents. This laboratory has developed an in vivo mutation assay that is based on the Pig-a locus. The Pig-a gene product is essential for the biosynthesis of glycosyl phosphatidylinositol (GPI) anchors. Mutations giving rise to nonfunctional GPI anchors prevent certain proteins from being expressed on the cell surface, and this represents a phenotype that can be measured by flow cytometry. The work proposed herein extends this line of investigation through an extensive inter-laboratory validation effort whereby rats will be treated with known mutagens and non-mutagens using both a short-term as well as a 28-day repeat dosing schedule. This assay validation effort will focus on an erythrocyte-based assay, a target cell population that has been studied most intensely to date. Concurrently, other work will be directed at developing means to measure Pig-a mutation in other tissues. Society will benefit from the proposed work as pharmaceutical and chemical companies are provided improved methods for conducting safety assessment work. PUBLIC HEALTH RELEVANCE: It is well known that DNA damage is a precursor to the development of cancer and other significant diseases. Therefore, it is in the interest of public health to reduce the occurrence of mutagenic chemicals in the environment, in our drugs, and from our workplaces. This research project will validate a powerful blood-based method that detects mutagenic agents, thereby enhancing the nation's ability to effectively reduce exposure to these toxic compounds. Additional work will be directed at developing mutant cell scoring methods that are compatible with other tissues.

描述（由申请人提供）：DNA突变是癌症出现的主要机制。这些事件也与诸如动脉粥样硬化和衰老等过程有关。因此，重要的是促进诱变研究的敏感分析方法，以及可以突变DNA的化学或物理剂的鉴定。目前存在测量体内突变的方法，每种方法都有自己的优势和局限性。虽然有些基于菌落形成并需要组织培养的工作，但有些则依靠昂贵的专有跨牙啮齿动物。该实验室开发了基于PIG-A基因座的体内突变测定法。 PIG-A基因产物对于糖基磷脂酰肌醇（GPI）锚的生物合成至关重要。引起非功能性GPI锚固的突变阻止某些蛋白在细胞表面表达，这代表了可以通过流式细胞仪测量的表型。本文提出的工作通过广泛的实验室间验证工作扩展了这一调查，从而使用已知的诱变剂和非毛ug虫使用短期和28天的重复给药时间表来治疗大鼠。这种测定验证工作将集中于基于红细胞的测定法，这是迄今为止最深入研究的目标细胞群。同时，其他工作将致力于开发用于测量其他组织中的猪A突变的方法。社会将从拟议的工作中受益，因为制药公司和化学公司提供了改进的进行安全评估工作的方法。 公共卫生相关性：众所周知，DNA损害是癌症和其他重大疾病的发展的先兆。因此，从环境，药物和工作场所中减少诱变化学物质的发生符合公共卫生的利益。该研究项目将验证一种强大的基于血液的方法，该方法检测诱变剂，从而增强了国家有效减少对这些有毒化合物的暴露的能力。其他工作将致力于开发与其他组织兼容的突变细胞评分方法。