Garbati Patrizia, Barbieri Raffaella, Cangelosi Davide, Zanon Carlo, Costa Delfina, Eva Alessandra, Thellung Stefano, Calderoni Matilde, Baldini Francesca, Tonini Gian Paolo, Modesto Paola, Florio Tullio, Pagano Aldo
IRCCS Ospedale Policlinico San Martino, 16132 Genova, Italy.
Laboratory of Molecular Biology, IRCCS Giannina Gaslini Institute, 16147 Genova, Italy.
Biomedicines. 2020 Nov 3;8(11):471. doi: 10.3390/biomedicines8110471.
To overcome the lack of effective pharmacological treatments for high-risk neuroblastoma (HR-NB), the development of novel in vitro and in vivo models that better recapitulate the disease is required. Here, we used an in vitro multiclonal cell model encompassing NB cell differentiation stages, to identify potential novel pharmacological targets. This model allowed us to identify, by low-density RT-PCR arrays, two gene sets, one over-expressed during NB cell differentiation, and the other up-regulated in more malignant cells. Challenging two HR-NB gene expression datasets, we found that these two gene sets are related to high and low survival, respectively. Using mouse NB cisplatin-treated xenografts, we identified two genes within the list associated to the malignant stage (MCM2 and carbonic anhydrase 9), whose expression is positively correlated with tumor growth. Thus, we tested their pharmacological targeting as potential therapeutic strategy. We measured mice survival and tumor growth rate after xenografts of human NB treated with cisplatin in the presence of MCM2/carbonic anhydrase 9 inhibitors (ciprofloxacin and acetazolamide). MCM2 or carbonic anhydrase 9 inhibition significantly increased cisplatin activity, supporting their possible testing for NB therapy.
为克服高危神经母细胞瘤(HR-NB)缺乏有效药物治疗的问题,需要开发能更好地模拟该疾病的新型体外和体内模型。在此,我们使用了一个包含神经母细胞瘤细胞分化阶段的体外多克隆细胞模型,以确定潜在的新型药物靶点。该模型使我们能够通过低密度逆转录聚合酶链反应(RT-PCR)阵列鉴定出两组基因,一组在神经母细胞瘤细胞分化过程中过度表达,另一组在恶性程度更高的细胞中上调。对两个HR-NB基因表达数据集进行分析,我们发现这两组基因分别与高生存率和低生存率相关。利用小鼠顺铂处理的神经母细胞瘤异种移植模型,我们在与恶性阶段相关的基因列表中鉴定出两个基因(微小染色体维持蛋白2(MCM2)和碳酸酐酶9),它们的表达与肿瘤生长呈正相关。因此,我们测试了将其作为潜在治疗策略进行药物靶向治疗的效果。在用顺铂处理人神经母细胞瘤的异种移植模型后,我们在存在MCM2/碳酸酐酶9抑制剂(环丙沙星和乙酰唑胺)的情况下测量了小鼠的生存率和肿瘤生长率。抑制MCM2或碳酸酐酶9可显著增强顺铂的活性,这支持了对它们进行神经母细胞瘤治疗测试的可能性。