PDPK1/AKT1通路异常与肺癌克唑替尼耐药及多激酶抑制剂克服耐药的机制研究

间变性淋巴瘤激酶（ALK）阳性的非小细胞肺癌（NSCLC）患者接受克唑替尼治疗后，耐药不可避免。我们对前期构建的克唑替尼耐药细胞系进行测序后发现其存在PDPK1基因的一种新转录本（PDPK1_n）及AKT1基因突变，通过蛋白结构分析推测PDPK1_n和AKT1突变可自发激活下游信号通路引发耐药；同时通过细胞和动物水平实验证明多激酶抑制剂CT-707可克服克唑替尼耐药，且FAK高表达和AKT1突变与其克服耐药有效性相关。因此，我们拟通过体内外实验研究PDPK1_n表达、AKT1突变和FAK拷贝数变异在肺癌克唑替尼耐药及多激酶抑制剂克服耐药中的作用机制，并在前瞻性临床试验的组织样本和液体活检标本中验证。研究将发现ALK抑制剂和多激酶抑制剂的新生物标志和潜在靶点，指导患者ALK抑制剂耐药后的治疗方案选择，拓展使用多激酶抑制剂克服ALK抑制剂耐药的新途径，为肺癌的个体化治疗提供实验依据和理论指导。

Non-small cell lung cancer (NSCLC) patients with anaplastic lymphoma kinase (ALK)-positive inevitably develop drug resistance after crizotinib treatment. We sequenced the previously established crizotinib-resistant cell lines and discovered a novel transcript of the PDPK1 gene (PDPK1_n) and AKT1 gene mutation. The protein structure analysis suggested that the PDPK1_n and AKT1 mutations can spontaneously activate signaling pathways, which lead to drug resistance. Of note, cell and animal experiments proved that multi-kinase inhibitor CT-707 can overcome crizotinib resistance, which is related to FAK expression and AKT1 mutations. Therefore, we aim to research the relationship of PDPK1_n expression, AKT1 mutations, FAK copy number variation and crizotinib resistance, overcoming drug resistance in vivo and vitro, and further validate in tumor tissue samples and liquid biopsy specimens in a prospective clinical trial. The study will identify new biomarkers and potential targets for ALK inhibitors and multi-kinase inhibitors, guide treatment for patients after resistance to ALK inhibitor , and expand the use of multi-kinase inhibitors. The results would also provide experimental and theoretical guidance for individualized treatment of lung cancer.