Institute of Clinical Pharmacology, School of Pharmaceutical Sciences, Sun Yat-Sen University, Guangzhou, 510060, People's Republic of China.
Department of Medical Oncology, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, People's Republic of China.
J Cancer Res Clin Oncol. 2021 Mar;147(3):725-737. doi: 10.1007/s00432-020-03476-4. Epub 2021 Jan 2.
Crizotinib is the first-line small molecule tyrosine kinase inhibitor for ALK-positive non-small cell lung cancer. In this study, a retrospective pharmacogenomics investigation was conducted to explore the relationship between genes related to RTK downstream signaling pathways and crizotinib-induced hepatic toxicity in ALK-positive NSCLC patients.
The variable importance analysis of random forest algorithm was applied to identify the significant features which contribute to the crizotinib sensitivity in Cancer Cell Line Encyclopedia (CCLE) database. The KEGG and reactome pathway enrichment analysis were conducted with EnrichR. The differential expression genes were identified with R package DESeq2 in CCLE liver derived cell lines between crizotinib sensitive and resistant groups. From 2012 to 2015, 42 NSCLC patients were enrolled in this study. 90 polymorphisms were genotyped using the Sequenom Massarray system. Sequencing of HGFR (c-Met) genes was carried out on the Ion Torrent Proton.
In total, 66.7% NSCLC patients suffered from crizotinib-induced liver toxicity and 11.9% progressed to severe hepatic toxicity. The features with the top importance from classification and regression random forest model were enriched in RTK downstream signaling pathways (JAK/STAT, RAS/RAF/MAPK, PI3K/AKT pathways) and immune system-related pathways. Collagen family genes (COL1A1, COL1A2, COL6A1, COL5A1) and other extracellular matrix protein (TNC, TAGLN, TENM2, EDIL3, VCAN, CNN1, SH3BP4, TAGLN), which were closely related to MAPK-ERK signaling pathways, were significantly enriched in crizotinib resistant cell lines. In multiple logistic regression, STAT1 rs10208033 (T > C) was significantly associated with crizotinib-induced liver toxicity (OR = 6.733, 95% CI 1.406-32.24, P = 0.017). Compared with non-CC, OR is 5.5 (95% CI 1.219-24.81, P = 0.027) for STAT1 rs10208033 CC genotype to develop crizotinib-induced liver toxicity. Further cell viability test in human fetal hepatocyte line, L-02, reveals that the STAT1 inhibitor might protect hepatocyte cells from the toxicity caused by crizotinib.
Polymorphism of rs10208033 is a potential biomarker for predicting crizotinib-induced hepatotoxicity. These results suggest that STAT1 plays an important role in crizotinib-induced hepatotoxicity. Further studies are needed to confirm our finding and understand the underlying mechanisms.
克唑替尼是治疗 ALK 阳性非小细胞肺癌的一线小分子酪氨酸激酶抑制剂。本研究通过回顾性药物基因组学研究，探讨与 RTK 下游信号通路相关的基因与 ALK 阳性 NSCLC 患者接受克唑替尼治疗后肝毒性的关系。
应用随机森林算法的变量重要性分析从癌症细胞系百科全书（Cancer Cell Line Encyclopedia，CCLE）数据库中识别出与克唑替尼敏感性相关的显著特征。应用 EnrichR 进行 KEGG 和 reactome 通路富集分析。在 CCLE 肝脏来源细胞系中，使用 R 包 DESeq2 对克唑替尼敏感和耐药组之间的差异表达基因进行鉴定。2012 年至 2015 年，共有 42 名 NSCLC 患者入组本研究。采用 Sequenom Massarray 系统对 90 个多态性进行基因分型。对 HGFR（c-Met）基因进行测序采用 Ion Torrent Proton。
共有 66.7%的 NSCLC 患者发生克唑替尼诱导的肝毒性，11.9%进展为严重肝毒性。分类和回归随机森林模型中最重要的特征富集于 RTK 下游信号通路（JAK/STAT、RAS/RAF/MAPK、PI3K/AKT 通路）和免疫系统相关通路。与 MAPK-ERK 信号通路密切相关的胶原家族基因（COL1A1、COL1A2、COL6A1、COL5A1）和其他细胞外基质蛋白（TNC、TAGLN、TENM2、EDIL3、VCAN、CNN1、SH3BP4、TAGLN）在克唑替尼耐药细胞系中显著富集。在多元逻辑回归中，STAT1 rs10208033（T > C）与克唑替尼诱导的肝毒性显著相关（OR = 6.733，95%CI 1.406-32.24，P = 0.017）。与非 CC 相比，STAT1 rs10208033 CC 基因型发生克唑替尼诱导的肝毒性的 OR 为 5.5（95%CI 1.219-24.81，P = 0.027）。在人胎肝细胞系 L-02 中进行的细胞活力检测表明，STAT1 抑制剂可能保护肝细胞免受克唑替尼引起的毒性。
rs10208033 多态性可能是预测克唑替尼诱导肝毒性的潜在生物标志物。这些结果表明 STAT1 在克唑替尼诱导的肝毒性中起重要作用。需要进一步的研究来证实我们的发现并了解潜在的机制。
