Wellcome Trust PhD Programme for Clinicians at the University of Dundee. (360G-Wellcome-090666_Z_09_Z)

Acute promyelocytic leukaemia is caused by the t(15; 17) genetic translocation resulting in the PML-RAR? fusion. This disrupts the function of these two genes, interfering with myeloid differentiation. The normal function of the promyelocytic leukaemia protein (PML) is not fully understood but it appears to have a role as a tumour suppressor. Through its use in a Chinese remedy, arsenic was noted to be extremely effective in the treatment of APL. This has subsequently been validated in clinical trials. The mechanism of its action is yet to be fully elucidated; however phosphorylation of PML is detected soon after arsenic treatment, and appears to precede small ubiquitin like (SUMO) modification which ultimately results in the degradation of PML by the proteasome. This research intends to use a small interfering (si) RNA screen to knock down kinase expression and assess changes in phenotype of the response of PML to arsenic treatment, using high content imaging of HeLa cells with yellow- fluorescent protein tagged PML and a synthetic lethal approach. Validation experiments will result in a list of kinases involved in the pathway and a subsequent bioinformatic analysis will aid definition of the pathway of arsenic mediated PML degradation.