Fig. 4: SA-lineage-related NB states are maintained across the metastatic dissemination.

a Illustration of patient samples from paired primary tumor (PT) and bone marrow (BM). b, c UMAP of 6,693 NB cells sub-sampled from 7 integrated paired PT/BM colored by transcriptomic clusters (b) and by tissue location (c). d Transcriptomic comparison of PT/BM clusters (in b) with NB SA-related clusters exposed to avian embryonic microenvironments. Similarity scores were computed with Spearman correlation across reference markers. e Bar plot of fractions of NB cells assigned to SA-related states per patient and per sample type. f UMAP of IGR-N91::GFP cells colored by cell location in E14 embryos -257 cells from SG tumors, 261 cells from ADR tumors, respectively 25 and 84 cells on PN and AOR, and 31 cells in the BM. g Bar plot with fractions of cells shown in (f) and assigned to SA-related states for each NB cell location. h Representative image of IGR-N91::GFP cells on PN showing heterogeneous expression of the c2 marker TOP2A in E14 embryos (observed in N = 3/3 embryos from 2 independent experiments). Sections were labeled with α-GFP, α-TUBB3, and α-TOP2A antibodies. i Heatmap of scaled expression values of marker genes extracted from genes differentially expressed between locations (two-sided Wilcoxon rank sum test; p-value < 0.05, log fold change > 0.1 min.pct > 0.1). j Representative images of IGR-N91::GFP cells in E14 embryos, in primary tumors or in contact with PN labeled with α-GFP, α-NF160 and α-PLXNB2 or α-ERBB4 antibodies. k, Gene set enrichment analysis from differentially expressed genes for each location. Pathways were extracted from ontology gene sets of molecular signature database (nperm = 1000, ranked by log fold change, abs(NES) > 1.3, Fischer’s exact test). l Bar plot showing pathway enrichment score (ES) (GO biological processes, p-value < 0.05) in NB cells migrating on PN or AOR based on differential gene expression (two-sided Wilcoxon rank sum test; p-value < 0.05, log fold change > 0.1, min.pct > 0.1). Square root of the ES is shown. Scale bars are indicated. Source data are provided as a source data file. Rb Rib, Sg Sympathetic ganglia.