Gamma-H2Ax quantification of low dose irradiation-induced DNA damage in patients receiving intensity modulated radiotherapy (IRMT)

The full text of the publication follows. Purpose/Objective: IMRT (Intensity Modulated Radiotherapy) offers greater target dose compliance yet may produce a comparative higher whole body dose. The aim of this study is to quantify γH2Ax foci in lymphocytes (an established marker of DNA double strand breaks) in patients undergoing IMRT. Material/Methods: Radical inverse planned IMRT was delivered to patients with brain tumours. Peripheral blood samples were collected from each patient at the following time points: baseline; weekly- prior to and 30 minutes after one treatment fraction; 2 and 6 weeks following completion of treatment. Whole blood was centrifuged to separate lymphocytes, which were fixed and stained for fluorescent immunocytochemistry. 150 cells per sample were visualized. γH2Ax foci were identified and counted using confocal microscopy. Results A low basal level of foci was present in all samples prior to any radiation exposure (0.233, SD 0.028). There was a significant increase in mean foci per cell in post radiotherapy treatment samples(0.367 foci per cell pre-treatment and 0.612 foci per cell post treatment, p=0.000) and no significant difference between post-treatment foci numbers at different times during treatment(for example, 0.518 foci per cell at week 1 and 0.760 at week 6, p=0.279). Mean foci numbers returned to background levels at 6 weeks following completion of radiotherapy (0.239 foci per cell at baseline and 0.219 foci per cell at 6 weeks, p=0.529). Comparison between patients treated with different delivery methods is ongoing. Conclusion: γH2Ax is a feasible marker of DNA damage in lymphocytes during IMRT. These data demonstrate a reproducible level of foci induction in patients undergoing IMRT for tumour targets in brain. There is no significant accumulation of foci during treatment and foci numbers return to baseline post treatment. This assay may be useful to assess differences in whole body dose when different delivery methods are used. (authors)