Iaia Ilenia, Gammaitoni Loretta, Cattaneo Giulia, Giraudo Lidia, Donini Chiara, Fiorino Erika, Primo Luca, Carnevale-Schianca Fabrizio, Aglietta Massimo, Puliafito Alberto, Sangiolo Dario
Candiolo Cancer Institute, FPO-IRCCS, 10060 Candiolo, Italy.
Department of Oncology, University of Turin, 10060 Candiolo, Italy.
Cancers (Basel). 2021 May 11;13(10):2302. doi: 10.3390/cancers13102302.
Cancer adoptive cell therapy (ACT) with HLA-independent tumor killer lymphocytes is a promising approach, with intrinsic features potentially addressing crucial tumor-escape mechanisms of checkpoint inhibitors. Cytokine-induced Killer (CIK) and Natural Killer (NK) lymphocytes share similar tumor-killing mechanisms, with preclinical evidence of intense activity against multiple solid tumors and currently testing in clinical studies. To improve the effective clinical translation of such ACT approaches, several fundamental questions still need to be addressed within appropriate preclinical contexts, capable of overcoming limitations imposed by most traditional two-dimensional assays. Here, we developed a novel experimental approach to explore, dissect, and visualize the interactions of CIK and NK lymphocytes with melanoma tumors in vitro in 3D. Primary melanoma cells were assembled into small tumors that were dispersed in a 3D matrix and challenged with patient-derived CIK or the NK-92 cell line. By means of imaging-based methods, we reported, visualized, and quantitatively measured the recruitment of CIK and NK on the 3D targets, their infiltration, and cytotoxic activity. Our results support the effective tumor recruitment and tumor infiltration by CIK and NK. Such features appeared dependent on the specific geometric aspects of the environment but can be explained in terms of directional migration toward the tumor, without invoking major feedback components. Overall, our 3D platform allows us to monitor the processes of tumor recruitment, infiltration, and killing by means of live measurements, revealing important kinetic aspects of ACT with CIK and NK against melanoma.
采用不依赖HLA的肿瘤杀伤淋巴细胞进行癌症过继性细胞疗法（ACT）是一种很有前景的方法，其内在特性可能解决检查点抑制剂关键的肿瘤逃逸机制。细胞因子诱导的杀伤细胞（CIK）和自然杀伤细胞（NK）具有相似的肿瘤杀伤机制，临床前证据表明它们对多种实体瘤具有强烈活性，目前正在进行临床研究测试。为了改善此类ACT方法的有效临床转化，在适当的临床前环境中仍需解决几个基本问题，以克服大多数传统二维检测方法所带来的局限性。在此，我们开发了一种新颖的实验方法，用于在三维体外探索、剖析和可视化CIK和NK淋巴细胞与黑色素瘤肿瘤的相互作用。将原发性黑色素瘤细胞组装成小肿瘤，分散在三维基质中，并用患者来源的CIK或NK-92细胞系进行攻击。通过基于成像的方法，我们报告、可视化并定量测量了CIK和NK在三维靶点上的募集、浸润及细胞毒性活性。我们的结果支持CIK和NK对肿瘤的有效募集和浸润。这些特性似乎取决于环境的特定几何方面，但可以用朝着肿瘤的定向迁移来解释，而无需引入主要的反馈成分。总体而言，我们的三维平台使我们能够通过实时测量来监测肿瘤募集、浸润和杀伤过程，揭示CIK和NK针对黑色素瘤进行ACT的重要动力学方面。
