Department of Radiology, University of California San Diego, La Jolla, California, USA
Celsense, Pittsburgh, Pennsylvania, USA.
J Immunother Cancer. 2023 Jun;11(6). doi: 10.1136/jitc-2023-007015.
Adoptive transfer of T cells is a burgeoning cancer therapeutic approach. However, the fate of the cells, once transferred, is most often unknown. We describe the first clinical experience with a non-invasive biomarker to assay the apoptotic cell fraction (ACF) after cell therapy infusion, tested in the setting of head and neck squamous cell carcinoma (HNSCC). A patient with HNSCC received autologous tumor-infiltrating lymphocytes (TILs) labeled with a perfluorocarbon (PFC) nanoemulsion cell tracer. Nanoemulsion, released from apoptotic cells, clears through the reticuloendothelial system, particularly the Kupffer cells of the liver, and fluorine-19 (F) magnetic resonance spectroscopy (MRS) of the liver was used to non-invasively infer the ACF.
Autologous TILs were isolated from a patient in their late 50s with relapsed, refractory human papillomavirus-mediated squamous cell carcinoma of the right tonsil, metastatic to the lung. A lung metastasis was resected for T cell harvest and expansion using a rapid expansion protocol. The expanded TILs were intracellularly labeled with PFC nanoemulsion tracer by coincubation in the final 24 hours of culture, followed by a wash step. At 22 days after intravenous infusion of TILs, quantitative single-voxel liver F MRS was performed in vivo using a 3T MRI system. From these data, we model the apparent ACF of the initial cell inoculant.
We show that it is feasible to PFC-label ~70×10 TILs (F-TILs) in a single batch in a clinical cell processing facility, while maintaining >90% cell viability and standard flow cytometry-based release criteria for phenotype and function. Based on quantitative in vivo F MRS measurements in the liver, we estimate that ~30% cell equivalents of adoptively transferred F-TILs have become apoptotic by 22 days post-transfer.
Survival of the primary cell therapy product is likely to vary per patient. A non-invasive assay of ACF over time could potentially provide insight into the mechanisms of response and non-response, informing future clinical studies. This information may be useful to developers of cytotherapies and clinicians as it opens an avenue to quantify cellular product survival and engraftment.
过继性 T 细胞转移是一种新兴的癌症治疗方法。然而，细胞转移后的命运通常是未知的。我们描述了第一个使用非侵入性生物标志物来检测细胞治疗输注后细胞凋亡分数 (ACF)的临床经验，该方法在头颈部鳞状细胞癌 (HNSCC) 中进行了测试。一名患有 HNSCC 的患者接受了自体肿瘤浸润淋巴细胞 (TIL) 的标记，这些 TIL 用全氟碳 (PFC) 纳米乳液细胞示踪剂标记。纳米乳液从凋亡细胞中释放出来，通过网状内皮系统清除，特别是肝脏的枯否细胞，肝脏的氟-19 (F) 磁共振波谱 (MRS) 用于无创推断 ACF。
从一名 50 多岁的患者的右扁桃体复发性、难治性 HPV 介导的鳞状细胞癌的肺转移灶中分离出自体 TIL。切除肺转移灶用于 T 细胞收获和扩增，使用快速扩增方案。在培养的最后 24 小时内，将扩增的 TIL 与 PFC 纳米乳液示踪剂共孵育，然后进行洗涤步骤。在静脉输注 TIL 后 22 天，使用 3T MRI 系统对患者进行了体内单容积肝脏 F MRS 检查。根据这些数据，我们对初始细胞接种物的表观 ACF 进行建模。
我们证明在临床细胞处理设施中可以一次性标记约 70×10 的 TIL（F-TIL），同时保持>90%的细胞活力和基于标准流式细胞术的表型和功能释放标准。基于肝脏的定量体内 F MRS 测量，我们估计在转移后 22 天，约 30%的过继转移 F-TIL 细胞发生凋亡。
每个患者的主要细胞治疗产品的存活率可能不同。随着时间的推移，对 ACF 的非侵入性检测可能会提供对反应和无反应机制的深入了解，为未来的临床研究提供信息。这一信息可能对细胞疗法的开发者和临床医生有用，因为它为量化细胞产品的存活和植入开辟了一条途径。
