Fig. 5: Genetic tracing of NB cells across the dissemination process allows mapping their physical trajectories in avian tissues. | Nature Communications

Fig. 5: Genetic tracing of NB cells across the dissemination process allows mapping their physical trajectories in avian tissues.

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

Fig. 5

a Parsimony tree showing the genetic relationships between NB cells located in adrenal tumors (orange arm) or sympathetic tumors (blue arm) and subclones located in the dissemination paths (AOR, PN) and in the bone marrow (BM). Genetic information was extracted from SMART-Seq2 single cell data obtained from E14 avian embryos grafted at E2 with IGR-N91::GFP cells (SMARTseq-A1 dataset, N = 9 embryos). Genetic proximity between cells was computed using single nucleotide polymorphisms and indel alterations of transcriptomic regions to build a cell-to-cell genetic divergence matrix based on variant allele frequency. b–q Representative images of the physical paths taken by IGR-N91::GFP cells in E14 embryos (8 embryos from 5 independent experiments analyzed). Tissue sections or whole embryos were labeled with α-GFP and α-NF160 or α-SMA antibodies. E: Embryonic day. b, c show IGR-N91 primary tumors located in the SG (observed in N = 6/8 embryos) (b, enlargement in b’) and in the ADR (observed in N = 6/8 embryos) (c, enlargement in c’). d, e show NB cells transiting to the PN (observed in N = 8/8 embryos) from a sympathetic tumor (d, enlargement in d’) and from an adrenal tumor (e, enlargement in e’). f–i show NB cell migration along nerves at distance from primary tumor sites (f) and reaching innervated bones (g–i) and the BM (observed in N = 4/8 embryos) (j). k–n show adrenal tumors transiting via the AOR (observed in N = 6/8 embryos). k (enlarged optical section in k’) illustrates the proximity of adrenal tumors with the AOR and l-n are transverse sections of the adrenal/aorta region. o–q show NB cells migration along blood vessels (o) at distance from adrenal tumors and reaching the bones (p) and the BM (q, optical section in q’). b–j, l–p are sections of embryos imaged with confocal microscopy. h–k, q were imaged with lightsheet microscopy. Scale bars are indicated. Source data are provided as a source data file. Adr Adrenal gland, Ao Aorta, Bm Bone marrow, Drg Dorsal root ganglia, Lg Lung, Lv Liver, Sg Sympathetic ganglia, V Vertebrae.

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