肺腺癌酪氨酸激酶抑制剂治疗耐药的逆转机制研究

The application of tyrosine kinase inhibitors (TKI) targeted epidermal growth factor receptor (EGFR) mutation and anaplastic lymphoma kinase (ALK) rearrangement has made breakthrough progress in the treatment of advanced lung adenocarcinoma, but drug resistance has become an emerging problem. In the previous work, we found that ALK-inhibitor crizotinib had obvious inhibitory effects on the growth of generated EGFR-TKI（icotinib）-resistant cell line. In addition, we also found that epigenetic regulative drug chidamide had inhibitory effects on the growth of EGFR-TKI resistant cell lines with different genetic background and ALK-positive cell lines. Based on these significance findings, with the use of chromatin immunoprecipitation sequencing (ChIP-Seq) and western-blot, this study intends to explore the reversal effects and mechanisms of crizotinib in overcoming icotinib-resistance, and novel ALK inhibitor CT-707 in overcoming crizotini-resistance; meanwhile, further to explore the mechanisms of chidamide in reversing EGFR-TKI resistance, in vitro cell lines and animal models. The results of this study are expected to provide experiment data and basis for exploring new treatment strategies in overcoming TKI-resistance in clinical treatment.

针对EGFR基因突变及ALK融合基因的酪氨酸激酶抑制剂（TKI）治疗晚期肺腺癌已经取得了突破性进展，但耐药问题仍无法回避。前期工作发现，ALK激酶抑制剂克唑替尼对我们已经构建成功的EGFR-TKI(埃克替尼)耐药细胞系具有明显的生长抑制作用。同时还发现，表观遗传调节剂西达本胺对不同遗传背景的EGFR-TKI耐药细胞系及ALK融合基因表达阳性的肺腺癌细胞系也有一定的生长抑制作用。基于以上重要发现，本研究拟通过染色质免疫沉淀-测序（ChIP-Seq）、蛋白印迹等技术，在细胞系及动物模型中探讨第一代ALK激酶抑制剂克唑替尼对埃克替尼耐药的逆转作用机制，新一代ALK激酶抑制剂CT-707克服克唑替尼耐药的效果及其分子机制，同时进一步从表观遗传学角度阐述西达本胺逆转EGFR-TKI耐药的分子机制。上述研究结果将为临床探索克服酪氨酸激酶抑制剂耐药的治疗策略提供有益的实验数据和依据。

本研究通过细胞系及动物模型详细阐述了克唑替尼逆转EGFR-TKI耐药的分子机制，且从表观遗传学角度分析了西达本胺对不同背景的肺腺癌细胞系的抗肿瘤效果及逆转EGFR-TKI耐药的分子机制，并分析了CT-707对克唑替尼耐药细胞系的抗肿瘤效果，并通过敲降及过表达技术进一步锁定CT-707及逆转克唑替尼耐药的作用机制。本课题研究结果发表SCI期刊文章 3篇，影响因子共8.5分，在课题研究期间，成功培养研究生2名,并以此课题研究结果为基础成功申请到2019年国自然面上项目1项。本课题研究结果将为临床探索克服酪氨酸激酶抑制剂耐药的治疗策略提供有益的实验数据和依据。