The main aim of the project is to investigate processes leading to destabilisation of cells after exposure to ionising irradiation. We will, in particular, concentrate on "non-targeted " phenomena such as bystander effect and genomic instability. The underlying hypothesis is that radiation induces a variety of "sensors" and "alarm signals " in cells, which could be expressed as bystander effect and genomic instability. The ultimate aim of the present study is to identify mechanisms, possible relationship and biological meaning of bystander effect and genomic instability. We plan to use three different protocols of irradiation: conventional broad field, medium transfer technique and microbeam irradiation (microbeam facility is available at Gray Cancer Institute).
We plan to use hTERT-BJ1 normal human fibroblasts and hTERT-RPE1 normal human epithelium cell lines which are telomerase-immortalised. This enables us to study effect of irradiation within a wide time frame. A variety of endpoints planned to be used in the present study. Generally, they could be divided into two major categories either related to cellular response (apoptosis, cytogenetic markers and protein expression) or signalling between cells (intercellular communication, expression of connexins). The study of non-targeted effects of radiation is important for understanding the dose- response mechanisms relevant to low-dose natural (i.e. due to Radon) irradiation in vivo and radiation protection issues.
The present project gives a unique opportunity to investigate mechanisms of genomic instability and bystander effect in two human cell lines with an infinite life span. Wide arrays of conventional and innovative techniques are planned to be used. The applicant would benefit from learning of new techniques such as protein expression assays, advanced molecular and cytogenetic markers of radiation response. On other hand, the applicant would be able to transfer his own unique expertise of working.