Department of Clinical Hematology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, People's Republic of China.
Key Laboratory of Artificial Micro- and Nano-structures of Ministry of Education, School of Physics and Technology, Wuhan University, Wuhan 430072, China.
Analyst. 2020 Jun 15;145(12):4138-4147. doi: 10.1039/d0an00110d.
Cancer immunotherapy has achieved great success in hematological cancers. However, immune cells are a highly heterogeneous population and can vary highly in clonal expansion, migration and function status, making it difficult to evaluate and predict patient response to immune therapy. Conventional technologies only yield information on the average population information of the treatment, masking the heterogeneity of the individual T cell activation status, the formation of immune synapse, as well as the efficacy of tumor cell killing at the single-cell level. To fully interrogate these single-cell events in detail, herein, we present a microfluidic microwell array device that enables the massive parallel analysis of the immunocyte's heterogeneity upon its interaction pairs with tumor cells at the single-cell level. By precisely controlling the number and ratio of tumor cells and T cells, our technique can interrogate the dynamics of the CD8+ T cell and leukemia cell interaction inside 6400 microfluidic wells simultaneously. We have demonstrated that by investigating the interactions of T cell and tumor cell pairs at the single-cell level using our microfluidic chip, details hidden in bulk investigations, such as heterogeneity in T cell killing capacity, time-dependent killing dynamics, as well as drug treatment-induced dynamic shifts, can be revealed. This method opens up avenues to investigate the efficacy of cancer immunotherapy and resistance at the single-cell level and can explore our understanding of fundamental cancer immunity as well as determine cancer immunotherapy efficacy for personalized therapy.
癌症免疫疗法在血液癌症方面已经取得了巨大的成功。然而，免疫细胞是一个高度异质的群体，在克隆扩增、迁移和功能状态方面变化很大，这使得评估和预测患者对免疫治疗的反应变得困难。传统技术只能提供治疗的平均群体信息，掩盖了个体 T 细胞激活状态、免疫突触形成以及肿瘤细胞杀伤的功效在单细胞水平上的异质性。为了详细地全面探究这些单细胞事件，我们在此提出了一种微流控微井阵列装置，该装置能够在单细胞水平上大规模平行分析免疫细胞与肿瘤细胞相互作用时的异质性。通过精确控制肿瘤细胞和 T 细胞的数量和比例，我们的技术可以同时探究 6400 个微流控井中 CD8+T 细胞和白血病细胞相互作用的动力学。我们已经证明，通过使用我们的微流控芯片在单细胞水平上研究 T 细胞和肿瘤细胞对的相互作用，可以揭示批量研究中隐藏的细节，例如 T 细胞杀伤能力的异质性、时变的杀伤动力学以及药物治疗诱导的动态变化。这种方法为在单细胞水平上研究癌症免疫疗法的疗效和耐药性开辟了道路，并可以深入了解癌症免疫的基本原理，以及确定癌症免疫疗法对个性化治疗的疗效。
