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PES2020-Prosj.etabl.støtte H2020

High Throughput Estimation of DNA Repair Capability

Awarded: NOK 21,219

Project to be established: Developing a novel diagnostic tool for assessing DNA repair capability. DNA repair is a crucially important player in preventing mutations and therefore cancer. It is also important as a factor in determining individual patients ' response to cancer therapy. We will develop a diagnostic tool for DNA repair assessment based on novel, high throughput, miniaturized, automated techniques. Till now, such a tool has not been available, since DNA repair assays have been very labour-inte nsive. Our tool will be based on the comet assay - a sensitive, well-established method for measuring DNA breaks. Electrophoresis of lysed, gel-embedded nuclei gives comet-like fluorescent images in which the relative tail fluorescence indicates the leve l of DNA damage. We have developed high throughput versions of the assay, and now plan to adapt these for measuring repair, using three distinct approaches: a cellular repair assay, in which cells are treated with DNA damaging agent, incubated, and remain ing damage measured at intervals; an in vitro assay, based on incubation of cell extract with DNA substrate containing specific lesions; and an assay for accumulated repair Intermediates when repair synthesis is blocked. The new tool will be validated aga inst existing methods, and applied in a pilot clinical study. Our aim is that the method should become available as a standard tool for use in hospitals, and also in human biomonitoring studies, for example monitoring DNA repair in subjects occupationally exposed to DNA damaging agents. Another possible application is in personalising healthcare. Challenges: Defining role of partners, allocating workpackages, agreeing finances. R&D challenges: Miniaturising the repair assay to make use of small numbers of cells; applying robotic technology to automate the assay; devising a rapid scoring method; validating the new methods in clinical studies.

Funding scheme:

PES2020-Prosj.etabl.støtte H2020