非小细胞肺癌中ALK基因融合的不同亚型及丰度对靶向治疗的影响及其机制研究

Over the past decade, the treatment of non-small cell lung cancer (NSCLC) has been revolutionized by the usage of tyrosine kinase inhibitors (TKIs) with the approach to the driven genetic events of the tumor. Since the first ALK TKI was approved by FDA, targeted therapy for ALK-fusion NSCLC has shown remarkable response in a series of clinical trials. However, the response magnitude and duration are heterogeneous in clinic. Based on our previous study, the variants of ALK-fusion were found to be associated with the survival of targeted treatment, although its mechanism was still unknown. In this work, we further investigate the responsiveness of various ALK-rearrangement variants and abundance to TKI by using NGS. We also explored the proliferation ability, migration ability, as well as the activation ability of downstream signaling pathways among different ALK-fusion variants, both in vitro and in vivo. Our study will provide the evidence for the heterogeneous response of TKI among ALK-fusion patients. It will also contribute to the understanding of the biological behavior among different ALK-fusion cells.

自ALK酪氨酸激酶抑制剂被批准用于治疗ALK融合的非小细胞肺癌以来，许多患者都从此靶向治疗中获益。然而，临床中发现，同样为ALK融合的患者，其靶向治疗获益却具有明显差异性。基于申请人前期工作，发现ALK融合的亚型与靶向治疗预后之间存在一定关联，然而其影响预后的机制未明。故本申请拟对ALK融合亚型及丰度对预后的影响及其机制进行进一步的体内及体外研究。本研究将：1.通过高通量NGS方法，明确ALK亚型及丰度与ALK靶向治疗预后之间的关系。2.通过建立离体细胞模型，探索不同ALK亚型对细胞增殖能力、迁移能力、及下游信号通路活性的影响；同时明确不同亚型对不同ALK抑制剂是否存在相同的响应差异。3.通过建立裸鼠肿瘤模型，验证不同ALK亚型肿瘤的形成及发展的差异，并探索对不同ALK抑制剂的响应差异。本研究将有助于对临床ALK靶向治疗异质性的理解，帮助提示ALK融合非小细胞肺癌病人的靶向治疗选择及预后。

约有5%-7%的非小细胞肺癌的驱动基因变异为ALK基因融合，并且能够从ALK酪氨酸激酶抑制剂克唑替尼的靶向治疗中获益。然而，自ALK酪氨酸激酶抑制剂被批准用于治疗ALK融合的非小细胞肺癌以来，我们发现，同样为ALK融合的患者，其靶向治疗获益却具有明显差异性。本研究针对中国非小细胞肺癌ALK融合的人群，描述了其临床病理参数、亚型分布、及预后分析，揭示了不同ALK融合亚型患者的预后差异。此外，本研究构建了不同亚型的过表达慢病毒，通过感染A549细胞系，探索了细胞模型及小鼠模型中不同亚型的ALK融合蛋白的表达量、对下游信号通路的影响，从而为进一步精准预测ALK融合肺癌患者的预后及耐药提供了进一步的思路。