Lineage origin and microenvironment shape neuroblastoma transcriptional state and plasticity

Neuroblastoma, a neural-crest-derived malignancy of the peripheral nervous system, is a devastating pediatric disease, characterized by high intra- and intertumoral heterogeneity. While expression of several tumor expression modules correlates with poor patient survival, the determinants of their emergence and plasticity remain elusive. Here, we systematically dissected neuroblastoma transcriptional heterogeneity and measured how tumor expression programs are determined by early developmental signaling versus local tumor environment. To achieve this, we combined single-cell transcriptomics with high-throughput lineage tracing and tumor cell transplantations in zebrafish models of high-risk neuroblastoma. We observed transcriptional programs determined by the cell of origin, including an ALK-activated state linked to poor disease prognosis in humans – in contrast to plastic states associated with physiological processes. Even lineage-determined tumor states can be reprogrammed upon exposure to a developmental signaling environment, indicating high plastic potential in vivo and a crucial role for the signals received in early tumorigenesis for tumor phenotype.