Development of an osteosarcoma model with MYCN amplification and TP53 mutation in hiPS cell- derived neural crest cells

Mesenchymal stem cell- or osteoblast-derived osteosarcoma is the most common malignant bone tumor. Its highly metastatic malignant phenotypes, which are often associated with a poor prognosis, have been correlated with the modulation of TP53- and cell-cycle-related pathways. MYC, which regulates the transcription of cell-cycle modulating genes, is used as a representative prognostic marker for osteosarcoma. Another member of the MYC oncoprotein family, MYCN, is highly expressed in a subset of osteosarcoma, however its roles in osteosarcoma have not been fully elucidated. Here, we attempted to create an in vitro tumorigenesis model using hiPSC-derived neural crest cells, which are precursors of mesenchymal stem cells, by overexpressing MYCN on a heterozygous TP53 hotspot mutation (c.733G > A; p.G245S) background. MYCN-expressing TP53 mutated transformed clones were isolated by soft agar colony formation, and administered subcutaneously into the periadrenal adipose tissue of immunodeficient mice, resulting in the development of chondroblastic osteosarcoma. MYCN suppression decreased the proliferation of MYCN-induced osteosarcoma cells, suggesting MYCN as a potential target for a subset of osteosarcoma treatment. Further, comprehensive analysis of gene expression and exome sequencing of MYCN-induced clones indicated osteosarcoma-specific molecular features, such as the activation of TGF-beta signaling and DNA copy number amplification of GLI1. The model of MYCN-expressing chondroblastic osteosarcoma was developed from hiPSC-derived neural crest cells, providing a useful tool for the development of new tumor models using hiPSC-derived progenitor cells with gene modifications and in vitro transformation.

Automated summary

In this study, an in vitro tumorigenesis model of chondroblastic osteosarcoma was successfully established using hiPSC-derived neural crest cells overexpressing MYCN and with a heterozygous TP53 hotspot mutation. Suppression of MYCN decreased the proliferation of MYCN-induced osteosarcoma cells, indicating MYCN as a potential target for a subset of osteosarcoma treatment. Comprehensive gene expression analysis and exome sequencing revealed osteosarcoma-specific molecular features, including activation of TGF-beta signaling and DNA copy number amplification of GLI1. This study provides a useful tool for the development of new tumor models using gene-modified hiPSC-derived progenitor cells.