Functional characterisation of the role of PPP2R3B in malignant melanoma. (360G-Wellcome-104076_Z_14_Z)

The Fellow will be based at the UCL Institute of Child Health, and the CRUK London Research Institute. The key finding underpinning this project is the association of a novel germline copy number change of the gene PPP2R3B and melanocytic neoplasia. Key goals: 1. Full characterisation of the human germline genotype-melanoma disease phenotype associations of PPP2R3B copy number abnormalities, using two accurately-phenotyped cohorts 2. Elucidation of the molecular mechanism of oncogenesis induc ed by PPP2R3B copy number change in vitro. shRNA knock-down and over-expression of PPP2R3B in melanoma cell lines using lentiviral vector-mediated transfection will be compared to mock-transfected controls. Functional assays will include RT-PCR and immunoblotting for downstream targets of the MAPK, p53 and PI3K/Akt pathways, cell viability/apoptosis assays and cell cycle stage assays. Identification of potential therapeutic candidates from a targeted siRNA screen and testing of the FDA-approv ed PP2A activator FTY720. 3. Modelling of PPP2R3B copy number changes in animal models to document in vivo effects. Both murine and zebrafish knock-down and transgenic overexpression models will be made, with assessment of MM development and MAPK pathway functional analysis. Promising therapeutic candidates from the cellular model screens will be tested in the animal models.