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Integrated multi-omic analysis of pediatric metastatic osteosarcoma reveals endothelial cell plasticity and lineage infidelity.

Burks, J.; Wu, Y.; Bhuvaneshwar, K.; Syed, N.; Jung, D.; Sayers, C. M.; Williams, D. O.; Daulatabad, S. V.; Malone, T.; Galindo, J.; Mendez, M.; Cotter, J.; Pavisic, J.; Mukouyama, Y.-S.; Shern, J. F.; Kaplan, R. N.; McEachron, T. A.

2026-08-12 cancer biology
10.64898/2026.08.11.744222 bioRxiv
Show abstract

While recent research has increasingly focused on the role of fibroblasts and macrophages in osteosarcoma, the tumor vasculature remains poorly understood, particularly in metastatic disease. To address this gap, we performed single-nuclei multi-ome (RNA+ATAC) sequencing on 24 human metastatic osteosarcoma specimens. We found that endothelial cells adopt a hybrid endothelial-mesenchymal state resembling endothelial-to-mesenchymal transition (EndMT) and that a subset of diploid endothelial cells expresses osteoblastic transcriptional profiles and gene regulatory networks (GRN). Joint copy-number analysis further identified osteosarcoma cells with endothelial transcriptional programs and GRNs, consistent with vascular mimicry. In vitro assays and syngeneic lineage-tracing experiments validated that tumor educated endothelial cells acquire osteoblast-like features. Together, these findings reveal substantial plasticity among endothelial and osteosarcoma cells in human and murine metastatic osteosarcoma, provide new insight into the how the metastatic microenvironment shapes the tumor vasculature, and challenge current models of osteosarcoma biology.

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