ESR15: “DNA damage and repair as diagnostic tools”

Norgenotech: DNA damage and repair as diagnostic tools

i. Objective of research: To develop new comet assay-based tools for precision medical diagnostics and early detection of cancer.

ii. Current state of the art: The comet assay is the method of choice for assessing DNA damage at the level of individual cells. It is sensitive, simple, economic, and – when performed on white blood cells - relatively non-invasive. Human biomonitoring studies have shown increased DNA damage associated with exposure to a wide range of environmental and occupational agents, as well as with various diseases including cancer. Oxidative stress is a factor in many human diseases. With a simple modification of the comet assay – inclusion of a digestion of the DNA with a specific DNA repair enzyme that recognizes oxidized bases and converts them to breaks – we can measure the effects of oxidative stress at the DNA level. Commonly used enzymes are Fpg, hOGG1 and Endo III. Elevated levels of oxidized bases are seen in patients with diabetes type II, Alzheimer’s, Parkinson’s, rheumatoid arthritis, hyperlipidemia, as well as in Down syndrome. Individual capacity to repair DNA damage is regarded as a marker of susceptibility to cancer. DNA repair capacity can be measured with the comet assay, but this requires substantial numbers of cells; thus miniaturized assays are needed.

iii. Research methodology and approach: We will develop and validate a set of diagnostic tools for simultaneous assessment of DNA damage, DNA repair and redox status, in the form of high throughput, platform based on the comet assay. Our novel assay system to assess genome instability at the individual cell level will have applications in assessing the likely response of cancer patients to therapy, evaluating the predictive value of DNA repair as a cancer risk factor, testing effects of dietary or pharmaceutical intervention and identifying individuals at risk from occupational or environmental exposure.

iv. Originality and innovative aspects of the ESR project: Our invention presents a technological and conceptual breakthrough resulting in a 10-fold increase of the overall assay turnover.

v. Integration of the ESR project to the overall research programme: Our ESR will collaborate with the Legube, Crosetto and Lingner groups on our available technologies for the simultaneous assessment of DNA damage, DNA repair and redox status in cell samples and with the LXRepair on the validation of complementary assays for DNA damage detection.