Abstract
Purpose
Ewing sarcoma (EwS) is an aggressive malignant neoplasm composed of small round cells. The heterogeneity and developmental trajectories of EwS are uncertain.
Methods
Single-cell RNA sequencing was performed on 4 EwS tumor tissue samples, and 3 transcriptional atlases were generated. K-nearest neighbor algorithm was used to predict the origin of tumor cells at single-cell resolution. Monocle2 package was used to perform pseudotime trajectory analysis in tumor cells. Differentially expressed genes were compared against those in all other clusters via the FindMarkers function, and then they were subjected to GO analysis using clusterProfiler package.
Results
Combined with the results of k-nearest neighbor algorithm and pseudotime trajectory analysis in tumor cells, we thought meningeal EwS originated from neural crest cells during epithelial to mesenchymal transition and simulated the process of neural crest cell lineage differentiation. But for perirenal EwS and spinal EwS, we hypothesized that after the neural crest cell lineage mutated into them, the tumor cells did not maintain the differentiation trajectory of neural crest cell lineage, and the development trajectory of tumor cells became chaotic. GO analysis results showed that interferon signaling pathway-related biological processes play an essential role in the tumorigenesis and tumor progression process of EwS, and among these biological processes genes, JAK1 gene up-regulated most significantly and highly expressed in all tumor cells. Ruxolitinib was used to explore the function of JAK1. Targeting JAK1 can promote apoptosis of EwS tumor cells, inhibit the migration and invasion of EwS tumor cells, and inhibit cell proliferation by inducing cell cycle S phase arrest.
Conclusion
EwS was derived from neural crest cell lineage with variable developmental timing of oncogenic conversion, and the JAK1 might be a candidate for therapeutic targets of EwS.
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Abbreviations
- EwS:
-
Ewing sarcoma
- NCSCs:
-
Neural-crest-derived stem cells
- MSCs: :
-
Mesenchymal stem cells
- scRNA-seq: :
-
Single-cell RNA sequencing
- UMI: :
-
Unique molecular identifier
- UMAP: :
-
Uniform manifold approximation and projection
- DEGs: :
-
Differentially expressed genes
- CNVs: :
-
Copy-number variations
- kNN: :
-
K-Nearest neighbor
- GO: :
-
Gene ontology
- PDX: :
-
Patient-derived xenografts
- Pre-EMT NCCs I–II: :
-
Pre-epithelial to mesenchymal transition neural crest cells I–II
- EMT NCCs::
-
Epithelial to mesenchymal transition neural crest cells
- NCCs: :
-
Neural crest cells after EMT process
- SNs: :
-
Sensory neurons
- SCPs: :
-
Schwann cell precursors
- Cycling CHCs: :
-
Cycling chromaffin cells
- Non-cycling CHCs: :
-
Non-cycling chromaffin cells
- NPPCs: :
-
Nucleus pulposus progenitor
- SCs: :
-
Satellite cells
- NTCs: :
-
Neural tube cells
- JAK1: :
-
Janus kinase-1
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Acknowledgements
We thank Xing-Yu Huo for editing the English text of a draft of this manuscript.
Funding
This work was supported by the Cyrus Tang Foundation (ZSBK0070), Shanghai Municipal Key Clinical Specialty (no. shslczdzk05703), National Natural Science Foundation of China (no. 82072782), the Shanghai Hospital Development Center (Grant No. SHDC12018X22), and the Children’s National Medical Center (Grant no. EK112520180301).
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This study was approved by the ethics committee of Fudan University Children’s Hospital.
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Hong, B., Li, Y., Yang, R. et al. Single-cell transcriptional profiling reveals heterogeneity and developmental trajectories of Ewing sarcoma. J Cancer Res Clin Oncol 148, 3267–3280 (2022). https://doi.org/10.1007/s00432-022-04073-3
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DOI: https://doi.org/10.1007/s00432-022-04073-3