In vivo transcriptome modulation after low dose of high energy neutron irradiation

Complete text of publication follows. Objective: This project aims to the identification of an hypothetical transcriptome modulation of mouse peripheral blood lymphocytes and skin after exposure to high energy neutron in vivo. Positive candidate genes isolated from mice in in vivo experiments will be selected and evaluated for both radioprotection issues dealing with cosmic ray exposure, and for biomedical issues mainly for low doses and non-cancer effects. Methods: High energy neutron irradiation is performed at the ENEA Frascati, neutron generator facilities (FNG), specifically dedicated to biological samples. FNG is a linear electrostatic accelerator that produces up to 1.0 x 1011 n/s 14 MeV neutrons via the D-T nuclear reaction. The dose-rate applied for this study is of 0.7 cGy/min. The functional genomic approach has been performed on six animals for each experimental points: un-irradiated; 20 cGy, 6 hours and 24 hours delayed time after exposure. Preliminarily, a pool of total RNA is evaluated on commercial micro-arrays containing large collections of mus musculus cDNAs. Statistical filtering and functional clustering of the data is carried out using dedicated software packages. Results: Candidate genes are selected on the basis of responsiveness to 20 cGy of exposure, with a defined temporal regulation. We plan to organize a systematic screen focused on genes responding to our selection criteria, in in vivo mouse experiments, and correlate their differential expression to the human counterparts. A specific cross species database will be created with all the functional information available in standardized format (MIAME: minimal information about micro-arrays experiments). Conclusions: A lack of information on in vivo experiments is still evident for low doses exposure, especially for neutron of cosmic interest. Individual susceptibility, extensive number of animals to be processed, lack of standardization methodologies are among problems to be solved for these studies. To this end, we pursue to define a pattern of expression related to tissues of pivotal interest for both health and biomarkers exposure. The project is partially funded by Italian Space Agency (ASI), MOMA Contract, 2006-2009.