PO.TB02.01 · 肿瘤生物学
用于表征实体瘤微环境中 CAR T 疗法的高通量成像方法
High-throughput imaging approaches to characterize CAR T therapies for solid tumor microenvironments
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
CAR T 疗法彻底改变了血液系统肿瘤的治疗，但在实体瘤中面临重大障碍，主要归因于肿瘤微环境。致密的细胞外基质（ECM）区域阻碍淋巴细胞的迁移和浸润，降低了治疗效果。为了模拟这些临床挑战，我们开发了一种高通量成像检测方法，以评估 CAR T 在三维（3D）ECM 包埋肿瘤球体中的细胞毒性。
上皮细胞黏附分子（EpCAM）通常在上皮性癌症中过表达，为 CAR T 疗法提供了一个具有临床相关性的靶点。将靶向 EpCAM 的 CAR T 细胞针对包埋于 ECM 中以模拟实体瘤结构的 T-47D 球体进行了测试。我们基于成像的方法揭示了剂量依赖性的细胞毒性以及 CAR T 细胞对球体的深度浸润，尽管与悬浮培养相比其细胞病变反应有所延迟。此外，与未经工程改造的 T 淋巴细胞相比，这种细胞毒性是 EpCAM 工程化 CAR T 细胞所特有的。重要的是，包埋条件下升高的 IC 50 值凸显了 ECM 对治疗动力学的影响。这些结果为优化 CAR T 设计和给药策略以克服微环境屏障提供了可操作的见解。通过在生理相关模型中量化迁移、浸润和杀伤，这项工作支持了改善 CAR T 在实体瘤中疗效的转化研究，并为下一代免疫疗法的临床开发路径提供了信息。
仅供研究使用。不用于诊断程序。
查看英文原文 English abstract
CAR T therapies have revolutionized hematologic cancer treatment but face significant barriers in solid tumors, primarily due to the tumor microenvironment. Dense extracellular matrix (ECM) regions impede lymphocyte migration and infiltration, reducing therapeutic efficacy. To model these clinical challenges, we developed a high-throughput imaging assay to evaluate CAR T cytotoxicity in three-dimensional (3D) ECM-embedded tumor spheroids.
The epithelial cell adhesion molecule (EpCAM), typically overexpressed in epithelial cancers, provides a clinically relevant target for CAR T therapy. EpCAM-targeted CAR T cells were tested against T-47D spheroids embedded in ECM to mimic solid tumor architecture. Our imaging-based approach revealed dose-dependent cytotoxicity and deep infiltration of spheroids by CAR T cells, despite delayed cytopathic responses compared to suspension cultures. Furthermore, this cytotoxicity is distinct to EpCAM-engineered CAR T cells when compared to nonengineered T lymphocytes. Importantly, elevated IC 50 values under embedded conditions underscore the impact of ECM on therapeutic kinetics. These results offer actionable insights for optimizing CAR T design and dosing strategies to overcome microenvironmental barriers. By quantifying migration, infiltration, and killing in physiologically relevant models, this work supports translational efforts to improve CAR T efficacy in solid tumors and informs clinical development pathways for next-generation immunotherapies.
For Research Use Only. Not for use in diagnostic procedures.
利益披露 Disclosure
E. Heimsath,
Agilent Technologies, Inc. Employment.
P. Held,
Agilent Technologies, Inc. Employment.
P. Brescia,
Agilent Technologies, Inc. Employment.
J. Clayton,
Agilent Technologies, Inc. Employment.