Neuroblastoma (NB) is a childhood cancer in sympathetic nervous system cells. NB exhibits cellular heterogeneity, with adrenergic and mesenchymal states displaying distinct tumorigenic potentials. NB is highly vascularized, and blood vessels can form through various mechanisms, including endothelial transdifferentiation, leading to the development of tumor-derived endothelial cells (TECs) associated with chemoresistance. We lack specific biomarkers for TECs. Therefore, identifying new TEC biomarkers is vital for effective NB therapies. A stiffness-based platform simulating human arterial and venous stiffness was developed to study NB TECs in vitro. Adrenergic cells cultured on arterial-like stiffness transdifferentiated into TECs, while mesenchymal state cells did not. The TECs derived from adrenergic cells served as a model to explore new biomarkers, with a particular focus on GB3, a glycosphingolipid receptor implicated in angiogenesis, metastasis, and drug resistance. Notably, the TECs unequivocally expressed GB3, validating its novelty as a marker. To explore targeted therapeutic interventions, nanoparticles functionalized with the non-toxic subunit B of the Shiga toxin were generated, because they demonstrated a robust affinity for GB3-positive cells. Our results demonstrate the value of the stiffness-based platform as a predictive tool for assessing NB aggressiveness, the discovery of new biomarkers, and the evaluation of the effectiveness of targeted therapeutic strategies.
神经母细胞瘤（NB）是一种发生于交感神经系统细胞的儿童恶性肿瘤。该肿瘤具有细胞异质性，其中肾上腺素能状态与间充质状态表现出不同的致瘤潜能。神经母细胞瘤具有高度血管化特征，其血管可通过多种机制形成，包括内皮转分化过程，这一过程可产生与化疗耐药相关的肿瘤源性内皮细胞（TECs）。目前我们尚缺乏针对TECs的特异性生物标志物，因此发现新的TEC生物标志物对开发有效的神经母细胞瘤治疗方案至关重要。本研究开发了一种模拟人体动脉与静脉硬度的基质刚度平台，用于体外研究NB TECs。实验发现，在模拟动脉硬度的基质上培养的肾上腺素能细胞可转分化为TECs，而间充质状态细胞则不具备此特性。利用肾上腺素能细胞来源的TECs作为研究模型，我们重点探索了GB3这一与血管生成、转移和耐药相关的鞘糖脂受体作为新型生物标志物的潜力。值得注意的是，TECs明确表达GB3，证实了其作为新型标志物的价值。为探索靶向治疗策略，我们构建了功能化修饰志贺毒素无毒亚基B的纳米颗粒，该颗粒对GB3阳性细胞表现出强亲和力。本研究结果证实了基于基质硬度的平台在评估神经母细胞瘤侵袭性、发现新型生物标志物以及评价靶向治疗策略有效性方面具有重要预测价值。
肿瘤(癌症)患者之家