Acoustic assembly of patient tumor organoids for modeling cancer immunity

项目摘要 对肿瘤和免疫系统之间的天然动态相互作用进行建模对于开发和测试至关重要 新的精确免疫治疗策略以及预测对创新癌症的临床反应 治疗，如免疫检查点阻断疗法。巨大的努力一直集中在 目前病人来源的癌症模型的发展，包括2D原代癌细胞培养，3D球体 和器官培养，以及患者来源的异种移植(PDX)。然而，这些模型不能再现 患者的天然癌症-免疫相互作用动态，主要是由于他们的低吞吐量，漫长的培养周期 (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。 我们在这里的总体目标是声学地组装新的患者器官，代表 患者肿瘤的微环境成分，以筛选免疫细胞渗透和细胞毒性 以高吞吐量和省时的方式进行动态调整。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个无支架的同型肿瘤球体。这个 提出的项目的目标是(1)利用患者肿瘤声学组装大量异型有机化合物 样本；(2)监测动态T细胞与声学工程患者器官的相互作用 使用我们的微流控高通量、延时单细胞成像方法的柱状阵列；以及(3)确定T 肿瘤细胞的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结果。首先，将开发一种新的声学有机体模型 以形成大量的异型患者肿瘤器质。其次，将利用该平台来研究T 免疫抑制微环境中细胞肿瘤的侵袭动力学和耗竭 病人肿瘤。第三，该平台将允许高通量和高效率地筛选代理(例如免疫 检查点抑制剂)，用于开发治疗实体瘤的新的癌症免疫治疗策略。

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.