Abstract A182: T-cell exhaustion assessment using a fully automated sequential chromogenic multiplex assay

Aurélie Collignon, Assil Benchaaben, Anna R Martirosyan, M. Duval, Emilie Bonzom, Emmanuel Prestat, C. Haond, J. Fieschi
{"title":"Abstract A182: T-cell exhaustion assessment using a fully automated sequential chromogenic multiplex assay","authors":"Aurélie Collignon, Assil Benchaaben, Anna R Martirosyan, M. Duval, Emilie Bonzom, Emmanuel Prestat, C. Haond, J. Fieschi","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-A182","DOIUrl":null,"url":null,"abstract":"Cancer immunotherapy by anti-PD-1 or anti-PD-L1 antibodies is now established as an efficient approach to restore the cytotoxic activity of exhausted T-cells. However, the strong and durable response observed in non-small cell lung cancer (NSCLC) patients is limited to a fraction of the treated population. Biomarkers, such as PD-L1 expression on tumor cells or tumor mutational burden, are approved as predictive markers of the response to anti-PD-1 or anti-PD-L1 antibodies. However, they are far from perfect to select patients eligible to immunotherapy. Since exhausted T-cells express more than one checkpoint inhibitor, it is hypothesized that an anti-PD1 antibody may not be sufficient to restore the activity of exhausted cells. Multiplex detection of the major immune checkpoints, i.e., PD-1, LAG-3 and TIM-3, on T-cells within the tumor microenvironment could, on one hand, predict the poor response to a single agent and could, on the other hand, predict the best combination of antibodies targeting more than one immune checkpoint. Here we present an automated sequential chromogenic multiplex assay allowing the assessment of the expression of PD-1, LAG-3 and TIM-3 on CD3+/CD8+ cells on a single FFPE tumor tissue section. Briefly, a tissue section is sequentially stained, digitized, unstained and restained with antibodies targeting the five markers. Images of the whole slide are then analyzed by digital pathology. First, a newly developed software is used to co-register the 5 virtual slides and perform colors deconvolution. Detection of positive cells is performed for each marker independently, using Indica Lab’s HALO software. Then, individual cells can be analyzed to identify complex phenotypes and assess their density on the entire tissue section. In addition, tissue segmentation tools are used to assess densities of exhausted T-cells expressing the major immune checkpoints in parenchyma, tumor stroma and invasive margin regions. The quantitative evaluation of T-cells’ exhaustion based on the expression of more than one immune checkpoint could improve patient stratification and lead to individualized combinations of immunotherapy agents. Citation Format: Aurelie Collignon, Assil Benchaaben, Anna Martirosyan, Matthieu Duval, Emilie Bonzom, Emmanuel Prestat, Christophe Haond, Jacques Fieschi. T-cell exhaustion assessment using a fully automated sequential chromogenic multiplex assay [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A182.","PeriodicalId":170885,"journal":{"name":"Regulating T-cells and Their Response to Cancer","volume":"20 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Regulating T-cells and Their Response to Cancer","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-A182","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Cancer immunotherapy by anti-PD-1 or anti-PD-L1 antibodies is now established as an efficient approach to restore the cytotoxic activity of exhausted T-cells. However, the strong and durable response observed in non-small cell lung cancer (NSCLC) patients is limited to a fraction of the treated population. Biomarkers, such as PD-L1 expression on tumor cells or tumor mutational burden, are approved as predictive markers of the response to anti-PD-1 or anti-PD-L1 antibodies. However, they are far from perfect to select patients eligible to immunotherapy. Since exhausted T-cells express more than one checkpoint inhibitor, it is hypothesized that an anti-PD1 antibody may not be sufficient to restore the activity of exhausted cells. Multiplex detection of the major immune checkpoints, i.e., PD-1, LAG-3 and TIM-3, on T-cells within the tumor microenvironment could, on one hand, predict the poor response to a single agent and could, on the other hand, predict the best combination of antibodies targeting more than one immune checkpoint. Here we present an automated sequential chromogenic multiplex assay allowing the assessment of the expression of PD-1, LAG-3 and TIM-3 on CD3+/CD8+ cells on a single FFPE tumor tissue section. Briefly, a tissue section is sequentially stained, digitized, unstained and restained with antibodies targeting the five markers. Images of the whole slide are then analyzed by digital pathology. First, a newly developed software is used to co-register the 5 virtual slides and perform colors deconvolution. Detection of positive cells is performed for each marker independently, using Indica Lab’s HALO software. Then, individual cells can be analyzed to identify complex phenotypes and assess their density on the entire tissue section. In addition, tissue segmentation tools are used to assess densities of exhausted T-cells expressing the major immune checkpoints in parenchyma, tumor stroma and invasive margin regions. The quantitative evaluation of T-cells’ exhaustion based on the expression of more than one immune checkpoint could improve patient stratification and lead to individualized combinations of immunotherapy agents. Citation Format: Aurelie Collignon, Assil Benchaaben, Anna Martirosyan, Matthieu Duval, Emilie Bonzom, Emmanuel Prestat, Christophe Haond, Jacques Fieschi. T-cell exhaustion assessment using a fully automated sequential chromogenic multiplex assay [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A182.
A182: t细胞耗竭评估使用全自动顺序显色多重分析
目前,通过抗pd -1或抗pd - l1抗体进行癌症免疫治疗是一种有效的方法,可以恢复耗尽的t细胞的细胞毒性活性。然而,在非小细胞肺癌(NSCLC)患者中观察到的强烈和持久的反应仅限于一小部分治疗人群。生物标志物,如肿瘤细胞上的PD-L1表达或肿瘤突变负荷,被批准作为抗pd -1或抗PD-L1抗体反应的预测标志物。然而,它们远不能完美地选择适合免疫治疗的患者。由于疲惫的t细胞表达不止一种检查点抑制剂,因此假设抗pd1抗体可能不足以恢复疲惫细胞的活性。多重检测肿瘤微环境中t细胞上的主要免疫检查点,即PD-1、LAG-3和TIM-3,一方面可以预测对单一药物的不良反应,另一方面可以预测针对多个免疫检查点的抗体的最佳组合。在这里,我们提出了一种自动顺序显色多重检测方法,可以评估单个FFPE肿瘤组织切片上CD3+/CD8+细胞上PD-1、LAG-3和TIM-3的表达。简单地说,一个组织切片依次染色,数字化,不染色和保留抗体针对这五个标记。然后用数字病理学分析整个切片的图像。首先,使用新开发的软件对5张虚拟幻灯片进行共配准,并进行颜色反卷积。使用Indica Lab的HALO软件,对每个标记物独立进行阳性细胞检测。然后,可以分析单个细胞以识别复杂表型并评估其在整个组织切片上的密度。此外,组织分割工具用于评估表达实质、肿瘤间质和侵袭性边缘区域主要免疫检查点的耗尽t细胞的密度。基于多个免疫检查点表达的t细胞衰竭的定量评估可以改善患者分层,并导致免疫治疗药物的个体化组合。引文格式:Aurelie Collignon, Assil Benchaaben, Anna Martirosyan, Matthieu Duval, Emilie Bonzom, Emmanuel Prestat, Christophe Haond, Jacques Fieschi。使用全自动顺序显色多重测定法评估t细胞耗竭[摘要]。第四届CRI-CIMT-EATI-AACR国际癌症免疫治疗会议:将科学转化为生存;2018年9月30日至10月3日;纽约,纽约。费城(PA): AACR;癌症免疫,2019;7(2增刊):摘要nr A182。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信