LAG-3 Protein Expression in Non-Small Cell Lung Cancer and Its Relationship with PD-1/PD-L1 and Tumor-Infiltrating Lymphocytes
LAG-3 Protein Expression in Non-Small Cell Lung Cancer and Its Relationship with PD-1/PD-L1 and Tumor-Infiltrating Lymphocytes
复制标题
DOI：
10.1016/j.jtho.2017.01.019
复制
发表时间：
2017-05-01
影响因子：
20.4
通讯作者：
Hirsch, Fred R.
中科院分区：
文献类型：
--
作者：
He, Yayi;Yu, Hui;Hirsch, Fred R.
简介：靶向程序性死亡1（PD-1）/程序性死亡配体1（PD-L1）检查点的免疫治疗在NSCLC患者中显示出有希望的疗效。淋巴细胞激活3基因（LAG-3）是另一个重要的检查点，其在NSCLC中的作用尚不清楚。在这项研究中，我们研究了淋巴细胞活化3（LAG-3）蛋白的表达;其与PD-1，PD-L1，和肿瘤浸润淋巴细胞（TILs）的相关性;以及其与NSCLC生存率的相关性。方法：通过免疫组织化学方法在55个NSCLC细胞系中评估LAG-3（EPR 4392 [Abcam，剑桥，MA]）蛋白的表达。通过免疫组织化学评价LAG-3、PD-1（NAT 105 [Cell Marque，Rocklin，CA]）和PD-L1（22 C3 [Dako，Carpenteria，CA]）蛋白表达，并对139例NSCLC患者手术切除标本中的TIL丰度进行评分。我们还验证了62例未经治疗的NSCLC患者的结果，并检测了LAG-3表达与EGFR和KRAS突变和棘皮动物微管相关蛋白样4基因（EML 4）-间变性淋巴瘤受体酪氨酸激酶基因（ALK）重排的相关性。而LAG-3在36例NSCLC患者的TIL上表达（25.9%）。60份患者样本（43.2%）在TIL上呈PD-1阳性，25份（18.0%）在肿瘤细胞上呈PD-L1阳性。LAG-3和PD-1在肿瘤细胞上均不表达。与腺癌相比，LAG-3在非腺癌中的TIL上过表达（p = 0.031）。TIL上的LAG-3表达与TIL上的PD-1（p < 0.001）和肿瘤细胞上的PD-L1（p = 0.041）显著相关，但与TIL百分比无关（p = 0.244）。使用logistic回归模型，当将非腺癌与腺癌进行比较，将PD-1阴性的TIL与PD-1阳性的TIL进行比较时，LAG-3的OR分别为0.320（95%置信区间[CI]：0.110-0.929）和4.364（95% CI：1.898-10.031）。TILs为LAG-3阴性与LAG-3阳性的患者的无复发生存期显著不同（1.91年[95% CI：0.76-3.06] vs 0.87年[95% CI：0.27-1.47] [p = 0.025]）。同样，TIL的LAG-3状态（阴性vs阳性）确实显著影响总生存期（OS）（3.04年[95% CI：2.76-3.32] vs 1.08年[95% CI：0.421.74] [p = 0.039]）。使用Kaplan-Meier分析，我们发现PD-L1阴性肿瘤细胞和LAG 3阴性TIL的患者比PD-L1或LAG-3阳性或PD-L1和LAG-3均阳性的患者具有更长的无复发生存期（2.09年[95% CI：0.90-3.28] vs 1.42年[95% CI：0.46-2.34] vs 0.67年[95% CI：0.00-1.45] [p = 0.007]）。在验证阶段，LAG-3的高表达也与TIL上PD-1（p = 0.016）和肿瘤细胞上PD L1（p = 0.014）的较高表达显著相关。LAG-3的表达与EGFR（p = 0.325）、KRAS突变（p = 1.000）和ALK融合（p = 0.562）无相关性。其在非腺癌中的表达较高，并与PD-1/PD-L1表达相关。LAG-3阳性或LAG-3和PD-L1均阳性与术后早期复发相关。LAG-3与不良预后有关。(C)2017年国际肺癌研究协会。爱思唯尔公司出版All rights reserved.
Introduction: Immunotherapy targeting the programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1) checkpoint has shown promising efficacy in patients with NSCLC. Lymphocyte activating 3 gene (LAG-3) is another important checkpoint, and its role in NSCLC is still not clear. In this study we investigated lymphocyte activing 3 (LAG-3) protein expression; its correlation with PD-1, PD-L1, and tumor-infiltrating lymphocytes (TILs); and its association with survival in NSCLC.Methods: The expression of LAG-3 (EPR4392 [Abcam, Cambridge, MA]) protein was assessed in 55 NSCLC cell lines by immunohistochemistry. LAG-3, PD-1 (NAT 105 [Cell Marque, Rocklin, CA]), and PD-L1 (22C3 [Dako, Carpenteria, CA]) protein expression was evaluated by immunohistochemistry, and TIL abundance was scored in 139 surgically resected specimens from patients with NSCLC. We also verified results in samples from 62 patients with untreated NSCLC and detected a correlation between LAG-3 expression and EGFR and KRAS mutation and echinoderm microtubule associated protein like 4 gene (EML4)-anaplastic lymphoma receptor tyrosine kinase gene (ALK) rearrangement.Results: LAG-3 was not expressed on any of the 55 NSCLC cell lines. However, LAG-3 was expressed on the TILs in 36 patients with NSCLC (25.9%). Sixty patient samples (43.2%) were positive for PD-1 on the TILs, and 25 (18.0%) were positive for PD-L1 on tumor cells. Neither LAG-3 nor PD-1 was expressed on the tumor cells. LAG-3 was overexpressed on the TILs in nonadenocarcinoma compared with in adenocarcinoma (p = 0.031). LAG-3 expression on TILs was significantly correlated with that of PD-1 on TILs (p < 0.001) and PD-L1 on tumor cells (p = 0.041) but not with TIL percentage (p = 0.244). With the logistic regression model, the ORs for LAG-3 were 0.320 (95% confidence interval [CI]: 0.110-0.929) and 4.364 (95% CI: 1.898-10.031) when nonadenocarcinoma was compared with adenocarcinoma and TILs that were negative for PD-1 were compared with those positive for PD-1. Recurrence-free survival was significantly different in patients whose TILs were LAG-3 negative as opposed to LAG-3-positive (1.91 years [95% CI: 0.76-3.06] versus 0.87 years [95% CI: 0.27-1.47] [p = 0.025]). Likewise, LAG-3 status of TILs (negative versus positive) did significantly affect overall survival (OS) (3.04 years [95% CI: 2.76-3.32] versus 1.08 years [95% CI: 0.421.74] [p = 0.039]). Using Kaplan-Meier analysis, we found that patients with both PD-L1-negative tumor cells and LAG 3 -negative TILs have longer recurrence-free survival than patients who are either PD-L1-or LAG-3-positive or both PD-L1- and LAG-3-positive (2.09 years [95% CI: 0.90-3.28] versus 1.42 years [95% CI: 0.46-2.34] versus 0.67 years [95% CI: 0.00-1.45] [p = 0.007]). In the verification stage, high expression of LAG-3 was also significantly correlated with higher expression of PD-1 on TILs (p = 0.016) and PD L1 on tumor cells (p = 0.014). There was no correlation between LAG-3 expression and EGFR (p = 0.325) and KRAS mutation (p = 1.000) and ALK fusion (p = 0.562).Conclusions: LAG-3 is expressed on TILs in tumor tissues of some patients with NSCLC. Its expression was higher in nonadenocarcinoma and correlated with PD-1/PD-L1 expression. LAG-3 positivity or both LAG-3 and PD-L1 positivity was correlated with early postoperative recurrence. LAG-3 was related to poor prognosis. (C) 2017 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.