Fig. 1: l NB transcriptomic plasticity across disease progression in an avian embryonic microenvironment. | Nature Communications

Fig. 1: l NB transcriptomic plasticity across disease progression in an avian embryonic microenvironment.

From: Neuroblastoma plasticity during metastatic progression stems from the dynamics of an early sympathetic transcriptomic trajectory

Fig. 1

a–f Representative lightsheet imaging of chick embryos grafted with IGR-N91::GFP cells at E2 and harvested at E5 (N = 11) (a) or E14 (N = 7) from 3 independent experiments (b–f) and labelled with α-NF160, or α-SMA, and α-GFP antibodies. E: Embryonic day. Optical sections at E5 in a’ and a”. b, c illustrate E14 rib cage with sympathetic ganglia tumor (in N = 5/7 embryos, enlargement in b’) and/or adrenal tumor (in N = 7/7, optical section c’). d, e show NB cells on nerves (in N = 4/7 embryos) (d), and on the aorta (in N = 7/7 embryos) (e). f illustrates bone invasion (in N = 7/7 embryos) with optical sections highlighting cortical (f’) and marrow (f”) metastases. Scale bars are indicated. g Overview of IGR-N91::GFP cell location harvested for scRNAseq. h Integration of 998 IGR-N91::GFP cells (E0 cells in culture, and from N = 22 E5 and N = 15 E14 embryos from 5 independent experiments) in a UMAP colored by timepoint. i Bar plot showing pathway enrichment score (ES) (Gene Ontology biological processes, p-value < 0.05) based on gene differential expression in E5 vs E0 and in E14 vs E5 (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. j UMAP of NB cells colored by transcriptomic state (c0, c1, c2) with Louvain clustering annotation, at the same resolution (res = 0.3) for E0 (162 cells), E5 (178 cells) and E14 (658 cells) stages. k UMAP of IGR-N91::GFP cells integrated overtime colored by transcriptomic clusters (c0, c1, c2). l Heatmap of scaled expression values of genes markers for clusters shown in k (two-sided Wilcoxon rank sum test; p-value < 0.05, log fold change > 0.25, min.pct > 0.1) grouped by phenotype. m, n Representative images of IGR-N91::GFP cells in culture and in sections of E5 (N = 10 embryos from 4 independent experiments) and E14 (N = 8 embryos from 5 independent experiments) embryos labeled with α-ATAD2 (c0), α-TOP2A (c2) and α-CHGB (c1) antibodies. Source data are provided as a source data file. Adr Adrenal gland, Ao Aorta, Bm Bone marrow, Cb Cortical bone, Lg Lung, No Notochord, NT Neural Tube, Sg Sympathetic ganglia, V Vertebrae.

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