Oncogenic mutations of ALK kinase in neuroblastoma
Oncogenic mutations of ALK kinase in neuroblastoma
复制标题
DOI：
10.1038/nature07399
复制
发表时间：
2008-10-16
期刊：
影响因子：
64.8
通讯作者：
Ogawa, Seishi
中科院分区：
文献类型：
--
作者：
Chen, Yuyan;Takita, Junko;Ogawa, Seishi
即使最近的治疗取得了进展，晚期神经母细胞瘤仍然是最难治的儿科癌症之一(1)。神经母细胞瘤具有多种遗传变化，包括高频率的 MYCN 扩增、1p36 和 11q 杂合性丧失以及 17q 遗传物质的获得，所有这些都与神经母细胞瘤的发病机制有关(2-5)。然而，可靠分子靶点的缺乏阻碍了针对神经母细胞瘤的有效治疗药物的开发。在此，我们表明，间变性淋巴瘤激酶 (ALK) 最初被鉴定为非霍奇金淋巴瘤亚型 (NPM-ALK)(6-8) 中的融合激酶，最近在肺腺癌 (EML4-ALK)(9,10) 中被鉴定为融合激酶，它也是晚期神经母细胞瘤中遗传改变的常见靶标。根据我们使用高密度单核苷酸多态性基因分型微阵列对 215 个原发性神经母细胞瘤样本中的遗传病变进行全基因组扫描 (11-14)，ALK 基因座（MYCN 基因座的着丝粒）被确定为拷贝数增加和基因扩增的反复靶点。此外，ALK 的 DNA 测序揭示了 215 个新鲜肿瘤中的 13 个 (6.1%) 和 24 个神经母细胞瘤衍生细胞系中的 8 个 (33%) 中存在 8 个新的错义突变。主要样本中除一个突变外（13 个中的 12 个）均在疾病的 3 - 4 阶段被发现，并且隐藏在激酶结构域中。与野生型激酶相比，突变激酶被自身磷酸化并表现出增加的激酶活性。他们能够转化 NIH3T3 成纤维细胞，如它们在软琼脂中的集落形成能力以及它们在裸鼠中形成肿瘤的能力所示。此外，我们证明通过 RNA 干扰下调 ALK 可抑制含有突变 ALK 的神经母细胞瘤细胞的增殖。我们预计我们的研究结果将为晚期神经母细胞瘤的发病机制提供新的见解，并且 ALK 特异性激酶抑制剂可能会改善其临床结果。
Neuroblastoma in advanced stages is one of the most intractable paediatric cancers, even with recent therapeutic advances(1). Neuroblastoma harbours a variety of genetic changes, including a high frequency of MYCN amplification, loss of heterozygosity at 1p36 and 11q, and gain of genetic material from 17q, all of which have been implicated in the pathogenesis of neuroblastoma(2-5). However, the scarcity of reliable molecular targets has hampered the development of effective therapeutic agents targeting neuroblastoma. Here we show that the anaplastic lymphoma kinase (ALK), originally identified as a fusion kinase in a subtype of non- Hodgkin's lymphoma (NPM-ALK)(6-8) and more recently in adenocarcinoma of lung (EML4-ALK)(9,10), is also a frequent target of genetic alteration in advanced neuroblastoma. According to our genome- wide scans of genetic lesions in 215 primary neuroblastoma samples using high- density single- nucleotide polymorphism genotyping microarrays(11-14), the ALK locus, centromeric to the MYCN locus, was identified as a recurrent target of copy number gain and gene amplification. Furthermore, DNA sequencing of ALK revealed eight novel missense mutations in 13 out of 215 (6.1%) fresh tumours and 8 out of 24 ( 33%) neuroblastoma- derived cell lines. All but one mutation in the primary samples ( 12 out of 13) were found in stages 3 - 4 of the disease and were harboured in the kinase domain. The mutated kinases were autophosphorylated and displayed increased kinase activity compared with the wild- type kinase. They were able to transform NIH3T3 fibroblasts as shown by their colony formation ability in soft agar and their capacity to form tumours in nude mice. Furthermore, we demonstrate that downregulation of ALK through RNA interference suppresses proliferation of neuroblastoma cells harbouring mutated ALK. We anticipate that our findings will provide new insights into the pathogenesis of advanced neuroblastoma and that ALK- specific kinase inhibitors might improve its clinical outcome.