NB, being a highly metastatic cancer, is one of the leading causes of cancer-related deaths in children. Increased disease recurrence and clinical resistance in patients with metastatic high-risk NBs (HR-NBs) result in poor outcomes and lower overall survival. However, the paucity of appropriate in vivo models for HR-NB metastasis has limited investigations into the underlying biology of HR-NB metastasis. This study was designed to address this limitation and develop suitable immunocompetent models for HR-NB metastasis. Here, we developed several highly metastatic immunocompetent murine HR-NB cell lines. Our newly developed cell lines show 100% efficiency in modeling experimental metastasis in C57BL6 mice and feature metastasis to the sites frequently observed in humans with HR-NB (liver and bone). In vivo validation demonstrated their specifically gained metastatic phenotype. The in vitro characterization of the cell lines showed increased cell invasion, acquired anchorage-independent growth ability, and resistance to MHC-I induction upon IFN-γ treatment. Furthermore, RNA-seq analysis of the newly developed cells identified a differentially regulated gene signature and an enrichment of processes consistent with their acquired metastatic phenotype, including extracellular matrix remodeling, angiogenesis, cell migration, and chemotaxis. The presented newly developed cell lines are, thus, suitable and promising tools for HR-NB metastasis and microenvironment studies in an immunocompetent system.
神经母细胞瘤作为一种高转移性肿瘤，是儿童癌症相关死亡的主要原因之一。转移性高危神经母细胞瘤患者疾病复发率增高及临床耐药性增强，导致预后不良和总体生存率降低。然而，由于缺乏合适的高危神经母细胞瘤转移体内模型，限制了对其转移潜在生物学机制的研究。本研究旨在解决这一局限性，开发适用于高危神经母细胞瘤转移研究的免疫健全模型。我们成功建立了多个具有高转移能力的免疫健全小鼠高危神经母细胞瘤细胞系。这些新开发的细胞系在C57BL6小鼠实验性转移模型中表现出100%的建模效率，并具有向高危神经母细胞瘤患者常见转移部位（肝脏和骨骼）转移的特征。体内验证实验证实了这些细胞系特异性获得的转移表型。体外表征显示，这些细胞系具有增强的细胞侵袭能力、获得锚定非依赖性生长特性，并在IFN-γ处理后表现出MHC-I诱导抗性。此外，通过RNA-seq分析新开发细胞系，我们发现了差异调控的基因特征以及与其获得性转移表型相一致的生物学过程富集，包括细胞外基质重塑、血管生成、细胞迁移和趋化作用。因此，本研究开发的新型细胞系为在免疫健全系统中进行高危神经母细胞瘤转移及微环境研究提供了合适且具有前景的研究工具。
肿瘤(癌症)患者之家