Joshua Goulding – Preclinical validation of DNA copy number driven non-mutated dependency genes RRM2 and BRIP1 as novel drug targets in neuroblastoma

Objectives

Investigation of zebrafish preclinical models for MYCN/RRM2 and MYCN/BRIP1-driven neuroblastoma formation and exploration of combination drugging targeting BRIP1/RRM2-ATR-CHK1 axis signalling in replicative stress response. Synergistic targeting of RRM2 and BRIP1 through combination drugging will be executed on standard cell lines, organoids and zebrafish xenotransplants and/or transgenic animals. Preclinical zebrafish models for MYCN versus MYCN/RRM2 and MYCN/BRIP1-driven neuroblastoma formation will be investigated in early hyperplastic lesions and full-blown tumours to investigate impact of dependency gene overexpression on proliferation, ssDNA levels (including EDU-seq), double strand DNA breaks, R-loops using immunostaining on tumour sections and altered gene expression and epigenetic marks using bulk and single cell RNA-seq and CUT&RUN for genome wide mapping of chromatin marks and finally telomere length using Q-FISH.

Expected Results

Novel mechanistic insights in the role of dependency genes on early tumour initiating events and tumour maintenance and leverage on precision oncology drug schemes for phase 1 clinical studies