Seoul National University Cancer Research Institute, Seoul, Republic of Korea.
Seoul National University Cancer Research Institute, Seoul, Republic of Korea; Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea; Medical Research Center and Department of Laboratory Medicine, Seoul National University College of Medicine.
Cytotherapy. 2019 Jun;21(6):603-611. doi: 10.1016/j.jcyt.2019.03.312. Epub 2019 Apr 20.
Treatment with tyrosine kinase inhibitors (TKIs) has improved the outcomes for patients with non-small cell lung cancer (NSCLC) harboring targetable driver mutations. However, acquired resistance to TKIs invariably develops within approximately 1 year of treatment by various mechanisms, including gatekeeper mutations, alternative pathway activation and histological transformations. Because immunotherapy is an option for patients with drug-resistant cancers, we generated several TKI-resistant NSCLC cell lines in vitro, and then evaluated the cytotoxicity of NK92-CD16 cells to these resistant cells.
TKI-resistant NSCLC cells (H3122CR1, H3122LR1, H3122CR1LR1, PC-9GR, PC-9ER, EBC-CR1 and EBC-CR2) were established from NCI-H3122 (EML4-ALK fusion), PC-9 (EGFR exon19 deletion) and EBC-1 (MET amplification) after continuous exposure to crizotinib, ceritinib, gefitinib, erlotinib and capmatinib. Expression of ligands for natural killer (NK) cell receptors and total EGFR were analyzed using flow cytometry. NK cytotoxicity and antibody-dependent cell-mediated cytotoxicity (ADCC) using anti-EGFR monoclonal antibody (mAb) cetuximab were measured using NK92-CD16 as effectors and detected using the Chromium-release assay.
We found that NK92-CD16 cells preferentially killed TKI-resistant NSCLC cells when compared with their parental NSCLC cells. Mechanistically, intracellular adhesion molecule 1 (ICAM-1) was up-regulated in the TKI-resistant NSCLC cells and patients' tumors, and the ICAM-1 up-regulated cancer cells lines were less susceptible to NK cytotoxicity by blocking ICAM-1. Moreover, NK92-CD16 cell-induced cytotoxicity toward TKI-resistant NSCLC cells was enhanced in the presence of cetuximab, an EGFR-targeting mAb.
These data suggest that combinational treatment with NK cell-based immunotherapy and cetuximab may be promising for patients with TKI-resistant NSCLC.
针对携带可靶向驱动突变的非小细胞肺癌（NSCLC）患者，酪氨酸激酶抑制剂（TKI）的治疗已经改善了结局。然而，由于各种机制，包括门控突变、替代途径激活和组织学转化，TKI 治疗的获得性耐药在大约 1 年内不可避免地发生。由于免疫疗法是耐药性癌症患者的一种选择，我们在体外生成了几种 TKI 耐药 NSCLC 细胞系，然后评估 NK92-CD16 细胞对这些耐药细胞的细胞毒性。
从 NCI-H3122（EML4-ALK 融合）、PC-9（EGFR 外显子 19 缺失）和 EBC-1（MET 扩增）中建立 TKI 耐药 NSCLC 细胞系（H3122CR1、H3122LR1、H3122CR1LR1、PC-9GR、PC-9ER、EBC-CR1 和 EBC-CR2），方法是连续暴露于克唑替尼、塞瑞替尼、吉非替尼、厄洛替尼和卡马替尼。使用流式细胞术分析自然杀伤（NK）细胞受体配体和总 EGFR 的表达。使用 NK92-CD16 作为效应细胞，通过 Chromium-release 测定法检测抗 EGFR 单克隆抗体（mAb）西妥昔单抗的 NK 细胞毒性和抗体依赖性细胞介导的细胞毒性（ADCC）。
与亲本 NSCLC 细胞相比，我们发现 NK92-CD16 细胞优先杀伤 TKI 耐药 NSCLC 细胞。从机制上讲，TKI 耐药 NSCLC 细胞和患者肿瘤中细胞间黏附分子 1（ICAM-1）上调，并且 ICAM-1 上调的癌细胞系通过阻断 ICAM-1 对 NK 细胞毒性的敏感性降低。此外，在存在 EGFR 靶向 mAb 西妥昔单抗的情况下，NK92-CD16 细胞诱导 TKI 耐药 NSCLC 细胞的细胞毒性增强。
这些数据表明，NK 细胞为基础的免疫疗法联合西妥昔单抗治疗可能对 TKI 耐药的 NSCLC 患者有希望。
