This project will validate two high throughput human blood-based DNA damage assays and develop them into commercial kits. The assays monitor types of damage associated with important human diseases. Whereas the PIG-A assay reports on gene mutation, the micronucleated reticulocyte (MN-RET) assay is responsive to chromosomal breaks and/or losses. The biomarkers are applicable to both humans and laboratory animals and will fulfill two important needs: extension of findings in laboratory animal models to direct studies in humans, and performance of well-controlled mechanistic laboratory studies to understand observations first made in humans. The assays utilize immunomagnetic separation prior to flow cytometry to dramatically enrich the relevant cell populations and thereby enhance assay precision and sensitivity. By providing simple-to-use kits with thoroughly documented reproducibility and inter-laboratory transferability, and validating the biomarkers for specific uses, researchers will have available tools with unprecedented efficiencies for comprehensively studying those factors that contribute to inter-individual differences in human DNA damage. Applications include the study of drug treatments, host and/or life-style factors that contribute to inter-individual differences in DNA damage and repair, exaggerated sensitivities to anti- neoplastic therapies, and population-based epidemiology studies of environmental exposures, including occupational exposures. The project benefits from a strong multidisciplinary team of internationally recognized scientists with a proven track record of successfully converting research advances into reliable commercial assay kits.
Public Health Relevance Statement: Project Narrative This project will validate two high throughput human blood-based DNA damage assays and develop them into commercial kits. The assays represent significant advances in the ability to directly monitor important types of DNA damage in humans, and their cross-species nature is ideal for supporting mechanistic laboratory studies when necessary. Validation work will include but is not limited to assessments of assay reproducibility within and among laboratories, validation of target cells, and identification of physiological and lifestyle factors that affect assay results. The proposed work also includes the evaluation of several important use cases for these kits, for example studying DNA damage resulting from chemotherapies, and also systemic inflammation.
Project Terms: Affect; Age; Aging; Aliquot; Antineoplastic Agents; arm; base; Benchmarking; Biological Assay; Biological Markers; Blood; Blood specimen; Blood Volume; Bloom Syndrome; Body mass index; C-reactive protein; Cancer Patient; Cells; chemotherapy; Chromosomal Breaks; Code; Congenital Abnormality; Data; Development; Disease; DNA Damage; DNA Repair; DNA Repair Disorder; Dose; Environmental Exposure; epidemiology study; Erythropoiesis; Evaluation; Exhibits; Family suidae; Fanconi's Anemia; Flow Cytometry; Folic Acid; Frequencies; Gene Mutation; Genes; Genomic Instability; genotoxicity; Genotype; Hematopoietic; Heritability; Human; human disease; human DNA damage; Human Pathology; human study; hydroxyurea; Immunomagnetic Separation; Individual; Inflammation; Inflammatory Bowel Diseases; Integration Host Factors; International; Investigation; Kinetics; Laboratories; Laboratory Animal Models; Laboratory Animals; Laboratory Study; lifestyle factors; Malignant Neoplasms; Measurement; Messenger RNA; Monitor; mouse model; multidisciplinary; Mus; Mutagens; Mutation; Nature; Newborn Infant; next generation; Nijmegen Breakage Syndrome; Obesity; Occupational Exposure; Operative Surgical Procedures; patient population; Patients; Performance; Pharmaceutical Preparations; Pharmacotherapy; Phase; phase 2 study; Phenotype; Physiological; Platinum; Play; Population; population based; Population Study; prototype; Race; Rattus; Reporting; Reproducibility; Research; Research Personnel; response; Reticulocytes; Rodent; Rodent Model; Role; Sampling; Scientist; sex; Sickle Cell Anemia; Small Business Innovation Research Grant; Source; specific biomarkers; Specimen; temozolomide; Testing; tool; TP53 gene; Treatment Factor; Validation; Work
