Acoustic assembly of patient tumor organoids for modeling cancer immunity

项目概要 模拟肿瘤和免疫系统之间的天然动态相互作用对于开发和测试至关重要 新的精准免疫治疗策略，以及预测对创新癌症的临床反应 治疗，例如免疫检查点阻断疗法。我们付出了巨大的努力 开发当前源自患者的癌症模型，包括 2D 原代癌细胞培养物、3D 球体 类器官培养物以及患者来源的异种移植物（PDX）。然而，这些模型无法复制 患者的天然癌症免疫相互作用动态，很大程度上是由于其低通量、较长的培养周期 （几周）、缺乏肿瘤微环境成分（例如免疫细胞）和/或支架 干扰 T 细胞迁移和细胞相互作用。 我们的总体目标是通过声学方式组装代表患者的新型类器官 患者肿瘤的微环境成分，以筛选免疫细胞浸润和细胞毒性 以高通量和省时的方式进行动态分析。我们的初步研究证明了声学 使用标准细胞系在一天内组装约 6,000 个无支架同型肿瘤球体。的 拟议项目的目标是（1）利用患者肿瘤以声学方式组装大量异型类器官 样品； (2) 监测 T 细胞与声学工程患者类器官的动态相互作用 使用我们的微流体高通量、延时单细胞成像方法的柱阵列； (3)确定T 细胞肿瘤动态浸润和细胞毒性或耗竭。 我们预计拟议的工作将产生三个成果。首先，将开发一种新颖的声学类器官模型 形成大量异型患者肿瘤类器官。其次，利用这个平台来学习T 细胞肿瘤浸润动态和免疫抑制微环境中的耗竭，密切模仿 患者肿瘤。第三，该平台将允许高通量和高效率地筛选药物（例如免疫药物） 检查点抑制剂）用于开发治疗实体瘤的新型癌症免疫治疗策略。

Project Summary Modeling the native dynamic interaction between tumor and immune system is crucial for developing and testing new precision immunotherapeutic strategies, as well as predicting clinical response to innovative cancer treatments, such as immune checkpoint blockade therapy. Tremendous efforts have been focused on the development of current patient-derived cancer models including 2D primary cancer cell cultures, 3D spheroid and organoid cultures, and patient-derived xenografts (PDX). However, these models fall short of reproducing patients’ native cancer-immune interaction dynamics, largely due to their low throughput, lengthy culture periods (several weeks), lack of tumor microenvironmental components (e.g. immune cells), and/or scaffolding that interferes with T cell migration and cellular interaction. Our overall objective here is to acoustically assemble novel patient organoids that represent the microenvironmental components of a patient’s tumor in order to screen immune cell infiltration and cytotoxicity dynamics in a high-throughput and time efficient manner. Our preliminary research demonstrated the acoustic assembly of about 6,000 scaffold-free homotypic tumor spheroids in one day using standard cell lines. The proposed project aims to (1) acoustically assemble a large number of heterotypic organoids using patient tumor samples; (2) monitor the dynamic T cell interaction with acoustically-engineered patient organoids trapped on a pillar array using our microfluidic high throughput, time-lapse single cell imaging approach; and (3) determine T cell tumor dynamic infiltration and cytotoxicity or exhaustion. We expect the proposed work will yield three outcomes. First, a novel acoustic organoid model will be developed to form a high number of heterotypic patient tumor organoids. Second, this platform will be employed to study T cell tumor infiltration dynamics and exhaustion in immunosuppressive microenvironments that closely mimic the patient tumor. Third, this platform will allow high-throughput and high-efficiency screening of agents (e.g. immune checkpoint inhibitors) for the development of novel cancer immunotherapy strategies to treat solid tumors.