Durante Valeria, Wittwer Alina, Theek Benjamin, Martinez-Osuna Manuel, Donnadieu Emmanuel, Hardt Olaf, Eckardt Dominik, Bosio Andreas, Schallenberg Sonja, Herbel Christoph
Research and Development Department, Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany.
Université Paris Cité, CNRS, Inserm, Institut Cochin, Equipe Labellisée Ligue Contre le Cancer, Paris, France.
PLoS One. 2025 Aug 8;20(8):e0327322. doi: 10.1371/journal.pone.0327322. eCollection 2025.
Up until present day, chimeric antigen receptor (CAR)-T cell therapy has only been approved for hematological malignancies, as CAR-T cells do not show comparable efficacy in solid tumors. Therefore, understanding the features of the tumor microenvironment (TME), is key to improve efficacy of adoptive cell therapies (ACTs) against solid tumors. In this context, robust workflows, which dissect the complex interactions between CAR-T cells and the TME are still lacking. To address this need, we have established an ex vivo workflow co-culturing tissue slices from patient tumor resections with CAR-T cells. The workflow is composed of assessing several complementary attributes, such as cytokine release via flow cytometry, quantification of cell infiltration into the tumor and assessment of the regions of the tissue slice the CAR-T cell infiltrate into by using the MACSima™ imaging cyclic staining technology. Using this workflow it is possible to observe the behavior of CAR-T cells within the tumor and its TME, their infiltration into distinct tumor compartments, as well as to dissect the underlying molecular mechanisms that drive T cell migration, thanks to MACSima™ multiplexing technology and its ability to image several markers at the same time. Assessment of ovarian carcinoma tissue slices revealed substantial release of specific cytokines and increased infiltration of T cells in the tumor areas when CAR-T cells were added to the tissue slices as compared to non-engineered T cells. The establishment of this novel approach will enable researchers to better characterize the interaction between CAR-T cells and the TME. Tissue slices present an intrinsic heterogeneity, which is indeed an advantage compared to other in vitro models but can turn itself into complex results interpretation. Therefore, we recommend that any conclusion derived from this assay should be verified with complementary models.
直到目前，嵌合抗原受体（CAR）-T细胞疗法仅被批准用于血液系统恶性肿瘤，因为CAR-T细胞在实体瘤中未显示出可比的疗效。因此，了解肿瘤微环境（TME）的特征是提高过继性细胞疗法（ACTs）治疗实体瘤疗效的关键。在这种情况下，仍缺乏能够剖析CAR-T细胞与TME之间复杂相互作用的强大工作流程。为满足这一需求，我们建立了一种体外工作流程，将患者肿瘤切除组织切片与CAR-T细胞共培养。该工作流程包括评估几个互补的属性，例如通过流式细胞术检测细胞因子释放、定量细胞向肿瘤内的浸润以及使用MACSima™成像循环染色技术评估CAR-T细胞浸润到的组织切片区域。利用该工作流程，可以观察CAR-T细胞在肿瘤及其TME中的行为、它们向不同肿瘤区域的浸润情况，并且由于MACSima™多重技术及其同时对多个标志物成像的能力，还能够剖析驱动T细胞迁移的潜在分子机制。对卵巢癌组织切片的评估显示，与未改造的T细胞相比，当将CAR-T细胞添加到组织切片中时，肿瘤区域会大量释放特定细胞因子且T细胞浸润增加。这种新方法的建立将使研究人员能够更好地表征CAR-T细胞与TME之间的相互作用。组织切片呈现出内在的异质性，这与其他体外模型相比确实是一个优势，但可能会导致复杂的结果解读。因此，我们建议从该检测得出的任何结论都应使用互补模型进行验证。
