Self-assembly of hybrid 3D cultures by integrating living and synthetic cells

Self-assembly is a fundamental property of living matter that drives the three-dimensional (3D) organization of cell collectives such as tissues and organs. Here, the co-assembly of synthetic and natural cells is leveraged to create hybrid living 3D cancer cultures. We demonstrate that synthetic cells based on droplet-supported lipid bilayers can establish artificial tumor immune microenvironments (ART-TIMEs), mimicking immunogenic signals within tumoroids formed by the cell line PANC-1 and eliminating the need to integrate complex living immune cells. Using the ART-TIME approach, we identify a co-signaling mechanism between PD-1 and other T cell-derived surface receptors as a driver in immune evasion of pancreatic ductal adenocarcinoma. Overall design: RNA-Seq profiling of WT PANC-1 tumoroids or hybrid tumoroids with integrated synthetic cells carrying various combinations of PD-1 and other surface receptors.

自组装（Self-assembly）是生命物质的固有属性，可驱动组织、器官等细胞集群形成三维（3D）结构。本研究通过合成细胞与天然细胞的共组装，构建混合活体三维癌症培养体系。研究证实，基于液滴支撑脂质双分子层的合成细胞能够构建人工肿瘤免疫微环境（Artificial Tumor Immune Microenvironments，ART-TIMEs），可模拟PANC-1细胞系形成的类肿瘤体内的免疫原性信号，且无需整合复杂的活体免疫细胞。借助ART-TIME方法，本研究揭示了PD-1（Programmed Death 1）与其他T细胞来源表面受体之间的共信号通路，该通路是胰腺导管腺癌免疫逃逸的关键驱动因素。实验设计概述：对野生型（Wild Type，WT）PANC-1类肿瘤体，或是整合了携带PD-1与其他表面受体不同组合的合成细胞的混合类肿瘤体开展RNA测序（RNA Sequencing，RNA-Seq）表达谱分析。