In Vitro Modelling of Neuroblastoma Development using Human Pluripotent Stem Cells

Neuroblastoma is one of the most common cancers that appear in early childhood. It affects 1 in 8000 live births and accounts for 12%-15% of all deaths in childhood related to cancer. It develops in the sympathetic nervous system and is thought to be derived from trunk neural crest (NC) cells or their sympathoadrenal derivatives during embryonic development. Tumours present different degrees of differentiation and common genetic aberrations associated with poor prognosis in neuroblastoma include amplification of the MYCN gene and chromosome 17q. Current popular models for examining the origins of neuroblastoma involve the use of transgenic mouse lines/primary explants or established human neuroblastoma-derived cell lines. Here I describe the development of a tractable human pluripotent stem cell (hPSC) differentiation in vitro system that allows the temporal dissection of the early events associated with neuroblastoma initiation. This involves the efficient generation of the cell types that give rise to neuroblastoma i.e. trunk NC and sympathoadrenal cells from hPSCs. I exploited this system to characterise hPSC lines that harbour neuroblastoma-relevant aberrations such as 17q amplification as well as a line I generated with an inducible MYCN overexpression cassette. I was able to highlight the impact of these aberrations on both cellular specification and cancer-related cellular dynamics along a sympathoadrenal trajectory. The overexpression of MYCN leads to both differentiation blockage and the presentation of cancer hallmarks, whereas 17q amplification presented an undocumented differential response to WNT signalling during trunk axial specification, leading to downstream differentiation defects. My data provides the basis for an in vitro model that might fully recapitulate the onset and progression of neuroblastoma in a physiologically relevant context.