DNA double strand breaks (DSB) are the most deleterious type of DNA damage induced by ionizing radiation and cytotoxic agents used in the treatment of cancer. When DSBs are formed, the cell attempts to repair the DNA damage through activation of a variety of molecular repair pathways. One of the earliest events in response to the presence of DSBs is the phosphorylation of a histone protein, H2AX, to form γH2AX. Many hundreds of copies of γH2AX form, occupying several mega bases of DNA at the site of each DSB. These large collections of γH2AX can be visualized using a fluorescence microscopy technique and are called ‘γH2AX foci’. γH2AX serves as a scaffold to which other DNA damage repair proteins adhere and so facilitates repair. Following re-ligation of the DNA DSB, the γH2AX is dephosphorylated and the foci disappear.
We have developed a contrast agent, 111In-anti-γH2AX-Tat, for nuclear medicine (SPECT) imaging of γH2AX which is based on an anti-γH2AX monoclonal antibody. This agent allows us to image DNA DSB in vitro in cells, and in in vivo model systems of cancer. The ability to track the spatiotemporal distribution of DNA damage in vivo would have many potential clinical applications, including as an early read-out of tumour response or resistance to particular anticancer drugs or radiation therapy.
The imaging tracer principle states that a contrast agent should not interfere with the physiology of the process being imaged. Therefore, we have investigated the influence of the contrast agent itself on the kinetics of DSB formation, repair and on γH2AX foci formation and resolution and now wish to synthesise these data into a coherent kinetic-dynamic model.