Aberrant activation of anaplastic lymphoma kinase (ALK) can cause sporadic and familial neuroblastoma. Using a proteomics approach, we identified Bruton's tyrosine kinase (BTK) as a novel ALK interaction partner, and the physical interaction was confirmed by co-immunoprecipitation. BTK is expressed in neuroblastoma cell lines and tumor tissues. Its high expression correlates with poor relapse-free survival probability of neuroblastoma patients. Mechanistically, we demonstrated that BTK potentiates ALK-mediated signaling in neuroblastoma, and increases ALK stability by reducing ALK ubiquitination. Both ALK^WT and ALK^F1174L can induce BTK phosphorylation and higher capacity of ALK^F1174L is observed. Furthermore, the BTK inhibitor ibrutinib can effectively inhibit the growth of neuroblastoma xenograft in nude mice, and the combination of ibrutinib and the ALK inhibitor crizotinib further enhances the inhibition. Our study provides strong rationale for clinical trial of ALK-positive neuroblastoma using ibrutinib or the combination of ibrutinib and ALK inhibitors.

中文翻译：

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布鲁顿的酪氨酸激酶增强了ALK信号传导，并成为神经母细胞瘤的潜在治疗靶标。

间变性淋巴瘤激酶（ALK）的异常激活可引起散发性和家族性神经母细胞瘤。使用蛋白质组学方法，我们确定了布鲁顿酪氨酸激酶（BTK）为新型的ALK相互作用伴侣，并且通过共免疫沉淀法证实了物理相互作用。BTK在神经母细胞瘤细胞系和肿瘤组织中表达。它的高表达与成神经细胞瘤患者的无复发生存率差有关。从机制上讲，我们证明了BTK增强了神经母细胞瘤中ALK介导的信号传导，并通过减少ALK泛素化来增加ALK稳定性。ALK ^WT和ALK ^F1174L均可诱导BTK磷酸化并提高ALK ^F1174L的容量被观察到。此外，BTK抑制剂依鲁替尼可有效抑制裸鼠中神经母细胞瘤异种移植的生长，并且依鲁替尼和ALK抑制剂克唑替尼的组合进一步增强了抑制作用。我们的研究为使用依鲁替尼或依鲁替尼和ALK抑制剂联合治疗ALK阳性神经母细胞瘤的临床试验提供了强有力的依据。