PD-1/PD-L1免疫抑制剂在非小细胞肺癌免疫治疗中的研究进展

摘 要：近年来免疫检查点领域研究进展迅速,众多新药相继出现,其中包括程序性细胞死亡蛋白1（programmed cell death protein 1,PD-1）单抗Pembrolizumab和Nivolumab,程序性细胞死亡蛋白配体1（programmed cell death protein ligand 1,PD-L1）单抗Atezolizumab。免疫检查点PD-1/PD-L1抑制剂给EGFR突变阳性非小细胞肺癌患者带来更多的生存获益,正逐渐改变国内外EGFR突变阳性晚期非小细胞肺癌的治疗模式。全文对PD-1/PD-L1为靶向的肿瘤免疫治疗在晚期EGFR突变阳性非小细胞肺癌患者治疗中的研究进展进行综述。     

外泌体PD-L1在非小细胞肺癌免疫治疗的研究进展

肿瘤免疫疗法在当前癌症治疗领域中愈来愈受瞩目,相关研究也层出不穷。对于非小细胞肺癌（non-small cell lung cancer, NSCLC）患者来说,近些年来,以程序性死亡受体1（programmed cell death 1, PD-1）/程序性死亡配体1（programmed cell death ligand 1, PD-L1）免疫抑制剂为代表的免疫检查点抑制剂（immunecheckpoint inhibitors, ICIs）已经成为治疗恶性肿瘤的一种最具前景的治疗方案。免疫检查点阻断治疗包括抗细胞毒T淋巴细胞相关抗原4（cytotoxic T lymphocyte-associated antigen 4, CTLA-4）单抗、抗PD-1单抗和抗PD-L1单抗,其中最被人熟知的为PD-L1免疫疗法。目前ICIs在临床治疗中取得了很不错的治疗效果,但有效率较低,因此我们希望获得更高的治疗有效率。近几年外泌体PD-L1在NSCLC免疫治疗中发挥了重要作用。本文就肿瘤外泌体PD-L1蛋白对肿瘤微环境的影响、预测免疫治疗效果以及作为NSCLC免疫治疗的新型治疗策略作一综...     

PD-1/PD-L1抑制剂在晚期非小细胞肺癌中的治疗进展

程序性死亡受体1(programmed death 1,PD-1)抑制剂Pembrolizumab进入一线正式标志着免疫检查点抑制剂在晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的治疗体系中占据了重要地位.临床试验结果证实PD-1/程序性死亡配体1(programmed death ligand 1,PD-L1)抑制剂在晚期NSCLC的一线、二线和多药耐药后治疗的疗效均要优于传统的化疗.一线使用Pembrolizumab联合化疗的客观有效率(objective response rate,ORR)最高可达80%;单药Pembrolizumab的无疾病进展时间(progression-free survival,PFS)接近1年(10.3个月),死亡风险比含铂双药化疗下降40%.单药Pembrolizumab、Nivolumab和Atezolizumab用于二线的疗效同样突出,总生存时间(overall survival,OS)可至1年左右.PD-L1的表达是PD-1/PD-L1抑制剂疗效的预测因子,在晚期NSCLC中阳性(≥1%)的比例约为60%左右,组织类型间差异不大,但是目前并无检测的金标准.     

PD-L1在非小细胞肺癌中的表达及相关性

目的 探讨程序性死亡配体-1(PD-L1)在非小细胞肺癌(NSCLC)中的表达与其临床病理参数的关系.方法 采用免疫组织化学两步法检测80例NSCLC组织中PD-L1的表达与其临床病理参数的关系.结果 NSCLC腺癌中PD-L1高表达28例(73.7％,28/38),鳞癌9例(34.6％,9/26),混合癌7例(43....     

多西他赛联合PD-1/PD-L1抑制剂二线治疗晚期非小细胞肺癌的临床分析

背景与目的 程序性死亡受体1(programmed cell death 1,PD-1)/程序性死亡配体1(programmed cell death ligand 1,PD-L1)抑制剂和多西他赛作为晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)患者的标准二线治疗方案,治疗效果有...     

PD-1和PD-L1治疗非小细胞肺癌的临床研究现状

程序性死亡受体1(PD-1)与其配体PD-L1属于CD28/B7家族,两者相互结合后能够调节肿瘤的微环境,使突变的细胞在变成无限增殖的肿瘤细胞之前被特异性清除,对免疫应答发挥负性调节的作用.目前研究表明,PD-1/PD-L1抑制剂在非小细胞肺癌(NSCLC)的临床应用中显示出巨大的潜力,成为继化疗、放疗、手术治疗和分子靶向治疗之后的又一大焦点.因此,本文对PD-1/PD-L1治疗NSCLC的临床研究现状及展望进行综述.     

非小细胞肺癌中与PD-L1和PD-L2表达相关的基因

背景程序性细胞死亡配体1(programmed cell death ligand-1,PD-L1)和配体2(programmed cell death ligand-2,PD-L2)与程序性细胞死亡受体1(programmed cell death protein-1,PD-1)的相互作用是一个介导免疫逃逸的免疫抑制检查点,因此是癌症中基于阻滞的免疫治疗的重要靶点。在非小细胞肺癌(nonsmall-celllungcancer,NSCLC)中,有必要对PD-1检查点阻滞反应生物学进行深入了解,并确定生物标志物以预测其对免疫疗法的临床反应。在本研究中,我们系统描述了NSCLC中PD-L1和PD-L2表达相关基因。方法我们进行了回顾性比较分析,来确定NSCLC中PD-L1和PD-L2 mRNA表达相关基因。为此,我们考察了肿瘤细胞系百科全书(Cancer Cell Line Encyclopedia,CCLE)数据库中肺–非小细胞(lung non-small-cell,Lung_NSC)和癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中肺腺癌(lung adenocarcinoma,LUAD)和鳞状细胞癌(lung squamous cell carcinoma,LUSC)的可用数据集。结果通过对CCLE数据集Lung_NSC的分析确定了PD-L1和PD-L2之间的表达相关性。此外,我们发现了489个基因与PD-L1相关、191个基因与PD-L2相关,以及111个基因与二者均有表达相关性。在TCGA数据集LUAD和LUSC中对PD-L1和PD-L2也进行了表达相关研究。在LUAD中,我们发现了257个基因与PD-L1相关、914个基因与PD-L2相关以及211个基因与二者均有表达相关性。在LUSC中,我们发现了26个基因与PD-L1相关、326个基因与PD-L2相关以及13个基因与二者均有表达相关性。只有少数基因表达在CCLE和TCGA数据集中均表现为与PD-L1和PD-L2相关。涉及干扰素信号转导基因的表达尤其与Lung_NSC中的PD-L1、LUSC中的PD-L2以及LUAD中的PD-L1和PD-L2的表达相关基因汇聚。在LUSC,PD-L1的表达,以及PD-L2的表达(相比之下相关程度较小)与染色体9p24区的基因相关,表明染色体9p24拓扑相关结构域是LUSC中PD-L1表达特别重要的驱动力。对PD-L1和PD-L2的受体PD-1、分化群80(cluster of differentiation,CD80)和排斥导向分子B(repulsive guidance molecule B,RGMB)的表达相关分析表明,在LUAD中PD-1和CD80表达与PD-L1和PD-L2均相关。在LUSC中CD80表达与PD-L2相关。结论我们提出了与NSCLC中PD-L1和PD-L2 mRNA表达相关的基因特征,这可能对于了解PD-1检查点阻滞反应生物学和开发基于基因特征的生物标志物以预测免疫疗法的临床反应具有重要意义。     

High expression of PD-L1 mainly occurs in non-small cell lung cancer patients with squamous cell carcinoma or poor differentiation

Background: Lung cancer is one of the most lethal cancers worldwide, but studies have shown that the higher the expression of programmed cell death protein 1 ligand 1 (PD-L1) in non-small cell lung cancer (NSCLC), the more likely it will benefit from anti-PD-L1 immunotherapy. The purpose of our study was to collect and analyze abundant clinical samples in order to provide evidence for clinicians and patients who might consider anti-PD-L1 immunotherapy while jointly formulating treatment plans. Methods: On the one hand, we obtained cases from The Cancer Genome Atlas (TCGA) database, including 498 lung squamous cell cancer (LUSC) patients and 515 lung adenocarcinoma (LUAD) patients. We studied the lung caner driver gene in LUSC and LUAD. On the other hand, PD-L1 expression was detected in lung cancer tissues of 1,008 NSCLC patients with immunohistochemistry staining (IHC), and we studied the correlation between PD-L1 protein expression and clinicopathological characteristics. Results: PD-L1 expression was higher in LUSC than in LUAD at the mRNA level. In univariate analysis, PD-L1 expression at the protein level was higher in patients who were males, were LUSC, were smokers, had a tumor diameter >3 cm, had poor differentiation, or had stages III~IV disease. In multivariate analysis, PD-L1 expression was higher in patients who were LUSC or in poor differentiation. Conclusion: In term of protein level, PD-L1 expression was higher in NSCLC patients who were LUSC or in poor differentiation. We recommend that PD-L1 IHC detection can be routinely performed in such populations that are likely to benefit most from PD-L1 immunotherapy.     

PI3K/AKT通路通过Nrf2途径调控PD-L1在非小细胞肺癌中的表达

背景与目的：程序性死亡［蛋白］配体-1（programmed death ligand-1,PD-L1）在肿瘤细胞中的表达对肿瘤逃避机体免疫监视具有重要的意义,其表达通常受MAPK、PI3K-AKT或STAT3等多条通路的影响,但这些通路具体经哪个关键环节尚不明确。拟通过PI3K/AKT信号通路对肺癌细胞A549、H460中PD-L1表达的调控具体机制进行探究。方法：使用shRNA技术选择性地沉默A549、H460细胞中的HEB、HTF4、Nrf2及FOXO3a等基因,构建缺陷细胞株;将目的基因PD-L1的3’UTR区域构建至pGL3-basic载体中,通过luciferase双报告基因体系检测PD-L1转录激活情况,并使用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR）技术验证细胞内PD-L1基因转录的情况;通过蛋白质印迹法（Western blot）检测细胞内PD-L1的总表达情况;免疫细胞与肿瘤细胞共培养,检测Nrf2基因敲除前后,人外周血单个核细胞（peripheral blood mononuclear cells,PBMC）对A549、H460细胞株的杀伤效果。结果：经10μg/mL的insulin刺激后,A549、H460细胞株中蛋白激酶B（AKT）基因磷酸化水平显著增强,伴随着PD-L1基因的转录水平和表达水平显著增强（P<0.001）,在HEB、HTF4及FOXO3a基因敲除的细胞株中,情况相似,而在Nrf2基因敲除的细胞株A549（A549 ^Nrf2- ）和H460（H460 ^Nrf2- ）细胞株中,PD-L1的转录和表达水平则与阴性对照组一样未见明显变化（P>0.05）;活性氧（reactive oxygen species,ROS）诱导剂isoproterenol能提高A549、H460细胞株中PD-L1基因的转录和表达水平,而ROS在被NAC解除后,PD-L1的转录和表达水平显著下调,进一步证明Nrf2对PD-L1基因的转录调控作用;wortmannin能逆转insulin刺激引起的AKT磷酸化水平增加,促进PD-L1基因的转录水平和表达水平下降,表明PD-L1的调控与AKT激活程度相关;在insulin和wortmannin的分别干预下,A549、H460细胞株中的Nrf2磷酸化水平能随着AKT磷酸化水平改变而改变,二者相关系数r分别为0.86和0.93,为强正相关;Nrf2敲除后,A549、H460细胞株与PBMC共培养后凋亡数量显著增加。结论：PI3K/AKT通路对肺癌细胞A549、H460中PD-L1表达具有关键调控作用,该调控是通过磷酸化激活Nrf2而实现的。     

Immunotherapy in the treatment of non-small cell lung cancer

Advances in the understanding of the role of the immune system in tumor immunosurveillance have resulted in the recognition that tumors can evade immune destruction via the dysregulation of co-inhibitory or checkpoint signals. This has led to the development of a generation immunotherapeutic agents targeting the immune checkpoint pathway. Recent early phase studies of immune checkpoint modulators, such as CTLA-4, PD-1 and PD-L1 inhibitors in NSCLC have reported promising results with prolonged clinical responses and tolerable toxicity. This article provides an overview of co-stimulatory and inhibitory molecules that regulate the immune response to tumors, recent therapies that have been developed to exploit these interactions and the role of predictive biomarkers in treatment selection.     

PD-1/PD-L1在肿瘤中的机制研究及其在甲状腺癌中的诊治价值

程序性死亡受体1(PD-1)和程序性死亡受体配体1(PD-L1)参与免疫检查点调节,与肿瘤的发生和发展密切相关。在甲状腺癌中,PD-L1的表达及PD-1阳性T细胞的增多可能预示着更高的侵袭性与更大的复发风险。抗PD-1/PD-L1治疗在许多肿瘤中已有显著疗效,但在甲状腺癌方面的疗效还比较局限,需要寻找能够更好预测疗效的生物标志物。进一步了解PD-1/PD-L1的作用机制及其在甲状腺癌中的诊治价值以及与其疗效相关的生物标志物,可为甲状腺癌患者提供新的治疗方法与思路。     

Comparison of Four PD-L1 Immunohistochemical Assays in Lung Cancer

Introduction

Four different programmed death ligand 1 immunohistochemical assays are approved or in development as companion or complementary diagnostics to different immunotherapeutic agents in lung carcinoma. We sought to determine whether these assays are technically equivalent and whether one antibody can be used on an alternate staining platform.

PD-1/PD-L1抑制剂治疗中晚期非小细胞肺癌疗效及安全性的Meta分析

目的:运用Meta分析方法系统评价PD-1/PD-L1抑制剂治疗中晚期非小细胞肺癌（NSCLC）的疗效及安全性。方法:检索CNKI、万方、Pubmed和Cochrane图书馆数据库,提取数据并进行核对,利用RevMan5.3软件的Cochrane偏倚风险工具,对随机对照试验进行风险评估,并应用该软件进行Meta分析。结果:共纳入7个随机对照试验,包括3 945例患者。Meta 分析结果显示:PD-1/PD-L1抑制剂在总生存期［HR=0.66,95%CI(0.61,0.72),P＜0.000 01］、无进展生存期［HR=0.72,95%CI(0.59,0.87),P=0.000 7］、总有效率［OR=1.91,95%CI(1.31,2.78),P=0.000 7］方面均高于化疗对照组;任何级别不良反应事件［OR=0.32,95%CI(0.25,0.40),P＜0.000 01］和3、4、5级不良反应事件［OR=0.23,95%CI(0.13,0.43),P＜0.000 01］低于化疗对照组。结论:在中晚期NSCLC的治疗中,PD-1/PD-L1抑制剂的疗效以及安全性均优于化疗药物。     

PD-1/PD-L1抑制剂与化疗治疗晚期非小细胞肺癌疗效的Meta分析

目的:本文旨在系统评价PD-1/PD-L1抑制剂对比化疗治疗非小细胞肺癌(non-small cell lung cancer,NSCLC)的有效性和安全性,采用Meta分析方法.方法:计算机检索Cochrane Library、PubMed、EM-Base、CNKI、万方数据库、VIP数据库,两名评价者独立评价纳入研究的质量、提取资料并交叉核对,运用Co-chrane量表评价纳入文献的方法学质量同质研究采用RevMan 5.3软件进行Meta分析.结果:共纳入5个随机对照试验,包括3042例病例.Meta分析结果显示:PD-1/PD-L1抑制剂相比较于对照组在总有效率[OR=1.58,95%CI(1.27,1.97),P<0.0001]、总生存期[HR=0.68,95%CI(0.62,0.75),P<0.00001]、无进展生存期[HR=0.79,95%CI(0.72,0.86),P<0.00001]高于对照组.在亚组分析中,PD-1/PD-L1抑制剂相比较于对照组在EGFR突变型的肺癌[HR=0.91,95%CI(0.76,1.11),P=0.35]中无明显差异,在EGFR野生型的肺癌[HR=0.67,95%CI(0.60,0.76),P<0.00001]中有差异.任何级别不良反应事件[OR=0.32,95%CI(0.27,0.39),P<0.00001]和3、4、5级不良反应事件[OR=0.18,95%CI(0.11,0.30),P<0.00001]低于对照组.结论:PD-1/PD-L1抑制剂方案治疗晚期NSCLC患者的疗效高于以多西他赛为主的化疗方案,且安全性优于后者.     

PD-L1 Immunohistochemistry Assays for Lung Cancer: Results from Phase 1 of the Blueprint PD-L1 IHC Assay Comparison Project

IntroductionThe Blueprint Programmed Death Ligand 1 (PD-L1) Immunohistochemistry (IHC) Assay Comparison Project is an industrial-academic collaborative partnership to provide information on the analytical and clinical comparability of four PD-L1 IHC assays used in clinical trials.MethodsA total of 39 NSCLC tumors were stained with four PD-L1 IHC assays (22C3, 28-8, SP142, and SP263), as used in the clinical trials. Three experts in interpreting their respective assays independently evaluated the percentages of tumor and immune cells staining positive at any intensity. Clinical diagnostic performance was assessed through comparisons of patient classification above and below a selected expression cutoff and by agreement using various combinations of assays and cutoffs.ResultsAnalytical comparison demonstrated that the percentage of PD-L1–stained tumor cells was comparable when the 22C3, 28-8, and SP263 assays were used, whereas the SP142 assay exhibited fewer stained tumor cells overall. The variability of immune cell staining across the four assays appears to be higher than for tumor cell staining. Of the 38 cases, 19 (50.0%) were classified above and five (13%) were classified below the selected cutoffs of all assays. For 14 of the 38 cases (37%), a different PD-L1 classification would be made depending on which assay/scoring system was used.ConclusionsThe Blueprint PD-L1 IHC Assay Comparison Project revealed that three of the four assays were closely aligned on tumor cell staining whereas the fourth showed consistently fewer tumor cells stained. All of the assays demonstrated immune cell staining, but with greater variability than with tumor cell staining. By comparing assays and cutoffs, the study indicated that despite similar analytical performance of PD-L1 expression for three assays, interchanging assays and cutoffs would lead to “misclassification” of PD-L1 status for some patients. More data are required to inform on the use of alternative staining assays upon which to read different specific therapy-related PD-L1 cutoffs.     

免疫检查点PD-1／PD-L1通路在小细胞肺癌中的作用

小细胞肺癌（SCLC）主要的特点为生长迅速且早期易发生广泛转移。尽管SCLC对于化疗和放疗敏感,但几乎所有的患者均在治疗后发生复发转移,预后差。免疫检测点抑制剂,尤其程序化细胞死亡受体-1（PD-1）／程序化细胞死亡配体-1（PD-L1）拮抗剂在SCLC的临床前和临床研究中均获得了良好效果,并且能够延长患者生存。免疫检测点疗法作为一种新兴的方法在未来可能会改变SCLC治疗模式。此外,有限的数据显示出PD-L1表达可能成为筛选获益人群一个有效的生物标记物。本文总结PD-L1作为标记物的发展历程,并同时阐述PD-1／PD-L1抑制剂在SCLC治疗中的进展。     

Pembrolizumab for the treatment of non-small cell lung cancer

Introduction: In the last years, a spectacular development of immunotherapeutic agents aimed at the PD-1/PD-L1 axis has taken place. This development of these checkpoint inhibitors has greatly influenced our approach to the treatment of lung cancer in first and second line. The limited toxicity profile and the ability to treat for prolonged periods, even in smokers, is a welcome expansion of the therapeutic arsenal of the oncologist.

Areas covered: This review highlights the results of recent clinical trials on pembrolizumab for the treatment of non-small cell lung cancer. The authors discuss both first and second line treatment with pembrolizumab as monotherapy and in combination therapies. Additionally, implications of the PD-L1 immunohistochemistry assay with the 22C3 antibody and its use in clinical practice and trials is discussed.

Expert commentary: A higher overall response, overall survival and a moderate toxicity profile is observed with the use of pembrolizumab, compared to chemotherapy, in both first and second line. These promising results have already translated into the registration of pembrolizumab in first and second line in patients with a high expression of PD-L1. However, as PD-L1 staining does not sufficiently discriminate responders from non-responders for all checkpoint inhibitors, there still is a need for a better predictive biomarker.