PO.IM01.05 · 免疫学
抗GPC3 CAR-T淋巴细胞有效靶向NSCLC，包括化疗耐药的干细胞样细胞
Anti GPC3 CAR T lymphocytes effectively target NSCLC, including chemoresistant stem like cells
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
目的：本研究旨在探索一种在肺癌背景下使用抗GPC3 CAR淋巴细胞进行基于细胞的免疫治疗的临床前策略，特别关注根除一个表现出干细胞样特征的化疗耐药肿瘤亚群。
程序：在一大系列鳞状非小细胞肺癌（NSCLC）手术标本和一组肿瘤细胞系中确认了GPC3表达。使用由健康供者以抗GPC3第二代CAR构建体工程化的CAR-T淋巴细胞进行功能测定。为可视化癌症干细胞（CSC）亚群，用慢病毒CSC检测载体转导肿瘤细胞系，其中GFP表达由干细胞基因启动子OCT4驱动，使具有干细胞样特征的细胞呈绿色。
结果：分别通过免疫组化和流式细胞术，在21/50例鳞状NSCLC样本和5种肿瘤细胞系（EBC1；H226；NCI-H23；NCI-H596；A-427）中确认了GPC3表达。抗GPC3 CAR-T淋巴细胞（平均表面CAR表达40%）有效杀伤所有测试的肿瘤细胞系（n = 5），在效靶比（E:T）分别为10:1和2:1时，平均特异性细胞毒活性范围为71%±13至42%±21（p<0.0001，对比未修饰T细胞对照），而对GPC3阴性的A-549细胞系无活性。该细胞毒活性完全涵盖了假定的癌症干细胞（CSC）区室（GFP⁺，此前显示其在体外对顺铂具有增强的耐药性），而且相比GFP⁻分化对应细胞对该亚群显示出优先杀伤，导致减少幅度约大两倍（p<0.05，E/T 5:1）。
结论：GPC3成为肺癌中基于细胞的免疫治疗的一个有效且有前景的靶点。尽管尚属初步，我们的数据支持这样的概念：抗GPC3 CAR-T细胞能够有效清除富含CSC的肿瘤区室，该区室被认为驱动化疗耐药和复发，从而为在晚期或复发无应答肺癌中基于细胞的免疫疗法的未来临床开发提供了依据。
查看英文原文 English abstract
Purpose: The aim of our study was to explore a preclinical strategy of cell-based immunotherapy using anti-GPC3 CAR-lymphocytes in the context of lung cancer, with a particular focus on eradicating a chemoresistant tumor subset displaying stem-like features.
Procedures : GPC3 expression was confirmed on a large series of surgical specimens from squamous non-small cell lung cancer (NSCLC) and on a panel of tumor cell lines. Functional assays were performed using CAR-T lymphocytes engineered from healthy donors with an anti-GPC3 2 nd generation CAR construct. To visualize the cancer stem cell (CSC) subset, tumor cell lines were transduced with a lentiviral CSC-detector vector in which GFP expression is driven by the stem-gene promoter OCT4, rendering green the cells with stem-like features.
Results: GPC3 expression was confirmed on 21/50 squamous NSCLC samples and 5 tumor cell lines (EBC1; H226; NCI-H23; NCI-H596; A-427), by immunohistochemistry and flow cytometry respectively. Anti-GPC3 CAR-T lymphocytes (mean surface CAR expression 40%) effectively killed all tested tumor cell lines (n = 5), with mean specific cytotoxic activity ranging from 71% ±13 to 42%±21 at effector-to-target (E:T) ratios of 10:1 and 2:1, respectively (p<0.0001 vs unmodified T cell controls), while showing no activity against the GPC3-negative A-549 cell line. The cytotoxic activity fully included the putative cancer stem cell (CSC) compartment (GFP⁺), previously shown with increased resistance to cisplatin in vitro, but also displayed preferential killing toward this subset compared with GFP⁻ differentiated counterparts resulting in approximately twofold greater reduction (p<0.05 , E/T 5:1).
Conclusions: GPC3 emerges as an effective and promising target for cell-based immunotherapy in lung cancer. Although preliminary, our data support the concept that anti-GPC3 CAR-T cells can effectively eliminate the CSC-enriched tumor compartment, which is believed to drive chemoresistance and recurrence, thus providing a rationale for future clinical development of cell-based immunotherapies in advanced or relapsed non-responsive lung cancer.
利益披露 Disclosure
L. Minori, None..
A. Proment, None..
F. Napoli, None.
L. Righi,
AstraZeneca ).
G. Doronzo, None..
E. Vigna, None.
K. Chaney,
AstraZeneca Employment, Stock.
W. Shim,
AstraZeneca Employment, Stock.
J. Stone,
AstraZeneca Employment, Stock.
P. Bironzo, None.
A. Hamilton,
AstraZeneca Employment, Stock.
S. Novello, None.
D. Sangiolo,
AstraZeneca ).