Ro Jooyoung, Kim Junyoung, Park Juhee, Choi Yongjun, Cho Yoon-Kyoung
Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 44919, South Korea.
Center for Algorithmic and Robotized Synthesis, Institute for Basic Science (IBS), Ulsan, 44919, South Korea.
Adv Sci (Weinh). 2025 Apr;12(13):e2410659. doi: 10.1002/advs.202410659. Epub 2025 Jan 13.
Current in vitro models of 3D tumor spheroids within the microenvironment have emerged as promising tools for understanding tumor progression and potential drug responses. However, creating spheroids with functional vasculature remains challenging in a controlled and high-throughput manner. Herein, a novel open 3D-microarray platform is presented for a spheroid-endothelium interaction (ODSEI) chip, capable of arraying more than 1000 spheroids on top of the vasculature, compartmentalized for single spheroid-level analysis of drug resistance, and allows for the extraction of specific spheroids for further analysis. As proof of concept, the crosstalk between breast cancer spheroids and vasculature is monitored, validating the roles of endothelial cells in acquired tamoxifen resistance. Cancer spheroids exhibited reduced sensitivity to tamoxifen in the presence of vasculature. Further analysis through single-cell RNA sequencing of extracted spheroids and protein arrays elucidated gene expression profiles and cytokines associated with acquired tamoxifen resistance, particularly involving the TNF-α pathway via NF-κB and mTOR signaling. By targeting the highly expressed cytokines (IL-8, TIMP1) identified, tamoxifen resistance in cancer spheroid can be effectively reversed. In summary, the ODSEI chip allows to study spheroid and endothelial interaction in various contexts, leading to improved insights into tumor biology and therapeutic strategies.
微环境中3D肿瘤球体的当前体外模型已成为理解肿瘤进展和潜在药物反应的有前途的工具。然而，以可控且高通量的方式创建具有功能性脉管系统的球体仍然具有挑战性。在此，提出了一种用于球体-内皮细胞相互作用（ODSEI）芯片的新型开放式3D微阵列平台，该平台能够在脉管系统上方排列1000多个球体，分隔用于单个球体水平的耐药性分析，并允许提取特定球体进行进一步分析。作为概念验证，监测了乳腺癌球体与脉管系统之间的串扰，验证了内皮细胞在获得性他莫昔芬耐药中的作用。在存在脉管系统的情况下，癌症球体对他莫昔芬的敏感性降低。通过对提取的球体进行单细胞RNA测序和蛋白质阵列的进一步分析，阐明了与获得性他莫昔芬耐药相关的基因表达谱和细胞因子，特别是通过NF-κB和mTOR信号传导涉及TNF-α途径。通过靶向鉴定出的高表达细胞因子（IL-8、TIMP1），可以有效逆转癌症球体中的他莫昔芬耐药性。总之，ODSEI芯片允许在各种情况下研究球体与内皮细胞的相互作用，从而更好地洞察肿瘤生物学和治疗策略。
