Acoustic assembly of patient tumor organoids for modeling cancer immunity

项目摘要 肿瘤与免疫系统之间的天然动态相互作用建模对于开发和测试至关重要 新的精确免疫策略，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。已经作出了巨大的努力， 开发当前患者来源的癌症模型，包括2D原代癌细胞培养物、3D球体 和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-免疫相互作用动力学，主要是由于其低通量，漫长的培养期 （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组装代表患者的新型类器官。 为了筛选免疫细胞浸润和细胞毒性， 以高吞吐量和时间有效的方式动态地进行。我们的初步研究表明， 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状体。的 拟议的项目旨在（1）利用患者肿瘤声学组装大量异型类器官 （2）监测动态T细胞与捕获在声学工程患者类器官上的动态T细胞相互作用。 柱阵列使用我们的微流体高通量，延时单细胞成像方法;和（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，将开发一种新的声学类器官模型 以形成大量异型患者肿瘤类器官。其次，利用该平台对T 在免疫抑制微环境中的细胞肿瘤浸润动力学和耗竭， 肿瘤患者第三，该平台将允许高通量和高效率地筛选试剂（例如免疫抑制剂）。 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.