A novel 4D ex vivo human lymphoma model to assess CAR T and NK cell efficacy

表达嵌合抗原受体(CARS)的T细胞和自然杀伤(NK)细胞过继转移治疗晚期B细胞恶性肿瘤已显示出显著的临床疗效。然而，许多患者并没有从这种治疗中受益，因为他们没有反应或随后复发。因此，了解肿瘤对CAR T/NK细胞的耐药机制具有重要意义。在启动临床试验之前，应该建立模型系统，在该模型系统中，可以对工程免疫细胞进行表征并测试其效力和安全性。到目前为止，很少有模型完美地概括了人类免疫系统和肿瘤微环境，一些模型揭示了CAR T/NK的局限性，这在其他模型中是矛盾的或完全忽略的。因此，仔细的模型选择是评估CAR T/NK细胞治疗的关键一步，也是癌症免疫治疗领域的一个主要问题。在这个项目中，我们将实施和优化过去几年由我们开发的创新的临床前人体模型，以实现对我们产生的CAR T/NK细胞疗效的高级监测。这些检测包括一个完全保留淋巴瘤环境的体外4D人体模型，以及一个能够分析工程T细胞和NK细胞运动的微通道系统。使用这两个模型，我们的目标是利用共聚焦荧光成像技术监测CD19 CAR T/NK细胞的早期和晚期反应以及它们杀死恶性细胞的能力。还将研究淋巴瘤环境中包含的抑制细胞和成分在CAR T/NK细胞反应中所起的作用。因此，我们相信，显著丰富当前技术水平的方法，以评估和预测特定的CAR免疫细胞为基础的疗法，并克服耐药机制。最后，在该项目期间获得的结果将使基于个性化免疫细胞的淋巴瘤治疗进一步优化。

Adoptive transfer of T and natural killer (NK) cells expressing chimeric antigen receptors (CARs) has shown remarkable clinical efficacy against advanced B cell malignancies. However, many patients do not benefit from this treatment either because they do not respond or subsequently relapse. It is therefore of paramount importance to understand the mechanisms of tumour resistance to CAR T/NK cells. Prior to initiating clinical trials, model systems in which engineered immune cells can be characterized and tested for their potency and safety should be in place. To date, few models perfectly recapitulate the human immune system and tumour microenvironment, and some models have revealed CAR T/NK limitations that were contradicted or missed entirely in other models. Thus, careful model selection is a crucial step in evaluating CAR T/NK cell treatment and a major issue in the field of cancer immunotherapy. In this project, innovative preclinical human models that have been developed over the last years by our will be implemented and optimized to enable advanced monitoring of CAR T/NK cell efficacy that we generate. These assays include an ex vivo 4D human model that fully preserves the lymphoma environment and a microchannel system that enables to dissect engineered T and NK cell motility. Using these two models, our objectives are to monitor, using confocal fluorescent imaging techniques, early and late CD19 CAR T/NK cell responses as well as their abilities to kill malignant cells. The role played by suppressive cells and components contained within the lymphoma environment on CAR T/NK cell responses will also be investigated. Thus, we believe to significantly enrich the current state of the art approaches for the evaluation and prediction of specific CAR immune cell-based therapeutics and to overcome resistance mechanisms. Finally, the results obtained during this project will allow further optimization of personalized immune cell-based therapy of lymphoma.