Purpose

Neuroblastoma, a common childhood tumor, remains one of the most elusive diseases to treat. To date, high-risk neuroblastoma is associated with low survival rates. To address this, novel and more effective therapeutic strategies must continue to be explored.

Methods

We employed a bioinformatics approach corroborated with in vitro and in vivo data. Samples from neuroblastoma patients were retrieved and immuno-stained for Bruton’s tyrosine kinase (BTK). To evaluate its effect on cellular functions, BTK expression in SK-N-BE(2) and SH-SY5Y neuroblastoma cells was downregulated using gene silencing or inhibition with ibrutinib or acalabrutinib. Xenograft mouse models were used to investigate the in vivo role of BTK in neuroblastoma tumorigenesis.

Results

We found that BTK was highly expressed in primary neuroblastoma samples, preferentially in MYCN-amplified neuroblastoma cases, and was associated with a poor prognosis. Immunohistochemical staining of tissues from our neuroblastoma cohort revealed a strong BTK immunoreactivity. We also found that neuroblastoma SK-N-BE(2) and SH-SY5Y cells were sensitive to treatment with ibrutinib and acalabrutinib. Pharmacologic or molecular inhibition of BTK elicited a reduction in the migratory and invasive abilities of neuroblastoma cells, and ibrutinib considerably attenuated the neurosphere-forming ability of neuroblastoma cells. Both inhibitors showed synergism with cisplatin. In vivo assays showed that acalabrutinib effectively inhibited neuroblastoma tumorigenesis.

Conclusions

From our data we conclude that BTK is a therapeutically targetable driver of neuroblastoma.

Graphical abstract

中文翻译：

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异常表达的布鲁顿酪氨酸激酶优先驱动神经母细胞瘤细胞中的转移性和干细胞样表型。

目的

神经母细胞瘤是一种常见的儿童肿瘤，仍然是最难以治疗的疾病之一。迄今为止，高危神经母细胞瘤与低存活率有关。为了解决这个问题，必须继续探索新的和更有效的治疗策略。

方法

我们采用了一种生物信息学方法，并得到了体外和体内数据的证实。取回神经母细胞瘤患者的样本并对布鲁顿酪氨酸激酶 (BTK) 进行免疫染色。为了评估其对细胞功能的影响，SK-N-BE(2) 和 SH-SY5Y 神经母细胞瘤细胞中的 BTK 表达被下调使用基因沉默或依鲁替尼或阿卡布替尼抑制。异种移植小鼠模型用于研究 BTK 在神经母细胞瘤肿瘤发生中的体内作用。

结果

我们发现 BTK 在原发性神经母细胞瘤样本中高度表达，尤其是在 MYCN 扩增的神经母细胞瘤病例中，并且与不良预后相关。来自我们神经母细胞瘤队列的组织的免疫组织化学染色显示出强烈的 BTK 免疫反应性。我们还发现神经母细胞瘤 SK-N-BE(2) 和 SH-SY5Y 细胞对 ibrutinib 和 acalabrutinib 治疗敏感。BTK 的药理学或分子抑制引起神经母细胞瘤细胞迁移和侵袭能力的降低，而依鲁替尼显着减弱了神经母细胞瘤细胞的神经球形成能力。两种抑制剂均显示出与顺铂的协同作用。体内试验表明，acalabrutinib 能有效抑制神经母细胞瘤的发生。

结论

根据我们的数据，我们得出结论，BTK 是神经母细胞瘤的治疗靶向驱动因素。

图形概要