Papageorgiou Savvas, Pashley Sarah L, O'Regan Laura, Straatman Kees R, Fry Andrew M
Department of Molecular and Cell Biology, University of Leicester, Lancaster Road, Leicester LE1 7RH, UK.
Advanced Imaging Facility, Core Biotechnology Services, University of Leicester, Leicester LE1 9HN, UK.
Cells. 2024 Nov 25;13(23):1954. doi: 10.3390/cells13231954.
The EML4-ALK oncogene drives tumour progression in approximately 5% of cases of non-small-cell lung cancers. At least 15 EML4-ALK variants have been identified, which elicit differential responses to conventional ALK inhibitors. Unfortunately, most, if not all, patients eventually acquire resistance to these inhibitors and succumb to the disease, which warrants the need for alternative targets to be identified. The most aggressive variant, EML4-ALK variant 3 (V3), assembles into a complex on interphase microtubules together with the NEK9 and NEK7 kinases, which leads to the downstream phosphorylation of NEK7 substrates. Overall, this promotes an elongated cell morphology and an enhanced migratory phenotype, which likely contributes to the increased metastasis often seen in V3 patients. Here, using two separate approaches to displace V3 from microtubules and a variety of in vitro assays, we show that microtubule association of EML4-ALK V3 is required for both V3 phenotypes, as removal of the oncogenic fusion protein from microtubules led to the dissociation of the V3-NEK9-NEK7 complex and the reversal of both phenotypic changes. Overall, we propose that targeting the interaction between EML4-ALK V3 and microtubules might offer a novel therapeutic option, independent of ALK activity, for V3+ NSCLC patients with acquired resistance to ALK inhibitors.
EML4-ALK致癌基因在约5%的非小细胞肺癌病例中驱动肿瘤进展。已鉴定出至少15种EML4-ALK变体，它们对传统ALK抑制剂产生不同反应。不幸的是，大多数（如果不是全部的话）患者最终会对这些抑制剂产生耐药性并死于该疾病，这就需要确定替代靶点。最具侵袭性的变体EML4-ALK变体3（V3）与NEK9和NEK7激酶一起在间期微管上组装成复合物，导致NEK7底物的下游磷酸化。总体而言，这促进了细胞形态的延长和迁移表型的增强，这可能导致V3患者中常见的转移增加。在这里，我们使用两种不同的方法将V3从微管上置换下来，并进行了各种体外试验，结果表明，EML4-ALK V3与微管的结合对于两种V3表型都是必需的，因为从微管上去除致癌融合蛋白会导致V3-NEK9-NEK7复合物的解离以及两种表型变化的逆转。总体而言，我们提出，针对EML4-ALK V3与微管之间的相互作用可能为对ALK抑制剂产生获得性耐药的V3+非小细胞肺癌患者提供一种独立于ALK活性的新型治疗选择。
