Pavesi Andrea, Tan Anthony T, Koh Sarene, Chia Adeline, Colombo Marta, Antonecchia Emanuele, Miccolis Carlo, Ceccarello Erica, Adriani Giulia, Raimondi Manuela T, Kamm Roger D, Bertoletti Antonio
Institute of Molecular and Cell Biology, Agency for Science, Technology, and Research, Singapore.
BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology, Singapore.
JCI Insight. 2017 Jun 15;2(12). doi: 10.1172/jci.insight.89762.
The tumor microenvironment imposes physical and functional constraints on the antitumor efficacy of adoptive T cell immunotherapy. Preclinical testing of different T cell preparations can help in the selection of efficient immune therapies, but in vivo models are expensive and cumbersome to develop, while classical in vitro 2D models cannot recapitulate the spatiotemporal dynamics experienced by T cells targeting cancer. Here, we describe an easily customizable 3D model, in which the tumor microenvironment conditions are modulated and the functionality of different T cell preparations is tested. We incorporate human cancer hepatocytes as a single cell or as tumor cell aggregates in a 3D collagen gel region of a microfluidic device. Human T cells engineered to express tumor-specific T cell receptors (TCR-T cells) are then added in adjacent channels. The TCR-T cells' ability to migrate and kill the tumor target and the profile of soluble factors were investigated under conditions of varying oxygen levels and in the presence of inflammatory cytokines. We show that only the 3D model detects the effect that oxygen levels and the inflammatory environment impose on engineered TCR-T cell function, and we also used the 3D microdevice to analyze the TCR-T cell efficacy in an immunosuppressive scenario. Hence, we show that our microdevice platform enables us to decipher the factors that can alter T cell function in 3D and can serve as a preclinical assay to tailor the most efficient immunotherapy configuration for a specific therapeutic goal.
肿瘤微环境对过继性T细胞免疫疗法的抗肿瘤疗效施加了物理和功能限制。对不同T细胞制剂进行临床前测试有助于选择有效的免疫疗法，但体内模型开发成本高昂且繁琐，而传统的体外二维模型无法重现靶向癌症的T细胞所经历的时空动态。在此，我们描述了一种易于定制的三维模型，其中可调节肿瘤微环境条件并测试不同T细胞制剂的功能。我们将人肝癌细胞以单细胞形式或作为肿瘤细胞聚集体纳入微流控装置的三维胶原凝胶区域。然后将经工程改造以表达肿瘤特异性T细胞受体的人T细胞（TCR-T细胞）添加到相邻通道中。在不同氧水平条件下以及存在炎性细胞因子的情况下，研究了TCR-T细胞迁移和杀伤肿瘤靶标的能力以及可溶性因子的概况。我们发现只有三维模型能够检测到氧水平和炎性环境对工程化TCR-T细胞功能的影响，并且我们还使用该三维微装置分析了免疫抑制情况下TCR-T细胞的疗效。因此，我们表明我们的微装置平台使我们能够解读可在三维空间中改变T细胞功能的因素，并可作为一种临床前检测方法，为特定治疗目标量身定制最有效的免疫治疗方案。
