It is well recognized that current batteries of genetic toxicology assays exhibit two critical deficiencies. First, the throughput capacity of in vitro mammalian cell genotoxicity tests is low, and does not meet current needs. Second, conventional assays provide simplistic binary calls, genotoxic or non-genotoxic. In this scheme there is little or no consideration for potency, and virtually no information is provided about molecular targets and mechanisms. These deficiencies in hazard characterization prevent genotoxicity data from optimally contributing to modern risk assessments, where this information is essential. We will address these major problems with current in vitro mammalian cell genetic toxicity assays by developing methods and associated commercial assay kits that dramatically enhance throughput capacity, and delineate genotoxicants' primary molecular targets, while simultaneously providing information about potency. Once biomarkers and a family of multiplexed assays have been developed for these purposes, an interlaboratory trial will be performed with prototype assay kits to assess the transferability of the methods.
Public Health Relevance Statement: Project Narrative DNA damage that cannot be faithfully repaired results in gene mutation and/or chromosomal aberrations, and these effects are known to contribute to cancer and other severe diseases. Thus, there is an important need for sensitive assays to evaluate chemicals for genotoxic and other deleterious effects. The work proposed herein will address issues that have plagued genotoxicity assessments for the last several decades: low throughput, lack of potency metrics, and little to no information about molecular targets. We will address these major problems with current genetic toxicity assays by developing new methods and associated commercial assay kits.
Project Terms: Address; Affect; Aneugens; Antioxidants; Appearance; aurora kinase; base; Benchmarking; Biological Assay; Biological Markers; Biological Response Modifiers; Bleomycin; Caspase; Cell Cycle; Cell Nucleus; Cells; Chemicals; Chromosome abnormality; Chromosomes; Classification; clastogen; Cleaved cell; Colcemid; Companions; Complex; Computer Simulation; computerized tools; Data; Data Analyses; Data Set; design; Disease; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA-PKcs; Dose; Epitopes; Etoposide; Exhibits; experimental study; Family; forest; GADD45A gene; Gamma-H2AX; Gene Mutation; Genetic; genotoxicity; Goals; Harvest; hazard; Histone H3; Human; In Vitro; inhibitor/antagonist; Intercalating Agents; Investigation; Kinetics; Label; Laboratories; Logistic Regressions; Machine Learning; Malignant Neoplasms; Mammalian Cell; Methods; Microtubules; Modeling; Modernization; Modification; Molecular Target; Mutagenicity Tests; next generation; NF-kappa B; Nuclear; Pathway interactions; Phase; Physiologic pulse; prediction algorithm; prevent; Procedures; Protocols documentation; prototype; Reagent; Reference Values; repaired; response; Risk Assessment; Schedule; Scheme; Series; Stains; targeted agent; Testing; Time; tool; Toxic effect; Toxicogenetics; TP53 gene; Training; Validation; virtual; Work
