Chen Yi-Wen, Chiang Huan-Jung, Liu Kuan-Ting, Kao Chun-Wei, Xie Shan-Ren, Su Chao-Ming, Shih Yu-Yin
Research and Development Center for x-Dimensional Extracellular Vesicles, China Medical University Hospital, Taichung, 404332, Taiwan.
Department of Bioinformatics and Medical Engineering, Asia University, Taichung, 41354, Taiwan.
Sci Rep. 2025 Aug 20;15(1):30582. doi: 10.1038/s41598-025-15846-4.
Traditional investigations of viral infection mechanisms have predominantly relied on two-dimensional (2D) cell culture models, which lack the structural organization and physiological relevance of native tissues. These systems often fail to capture key features such as spatial cell-cell interactions, tissue-specific heterogeneity, and microenvironmental complexity that govern virus-host dynamics in vivo. To address these limitations, we established an integrative platform that combines the strengths of both 2D and three-dimensional (3D) models to investigate the role of six-transmembrane epithelial antigen of prostate 3 (STEAP3), a membrane ferrireductase, in regulating viral infection in human intestinal epithelium. The 2D system enabled high-resolution mechanistic interrogation of STEAP3-dependent viral entry processes, while the patient-derived 3D colon organoid model recapitulated the architectural and cellular complexity of intestinal tissue, allowing spatially resolved assessment of infection patterns. Using this integrated approach, we found that STEAP3 knockdown significantly increased viral entry and infection, particularly in enterocytes and enteroendocrine cells. To further mimic physiological conditions in human body, we developed a vascularized organoid-on-chip model, in which increased viral signals were observed within vascular lumens upon STEAP3 depletion, suggesting a protective role of STEAP3 in limiting viral dissemination. For efficient and multiplexed screening of antiviral mechanisms, we also fabricated a 3D-printed 27-well chip tailored for organoid culture. By leveraging the complementary advantages of both 2D and 3D systems, this study demonstrates the power of integrated biomimetic modeling platforms to investigate antiviral defense mechanisms and underscores their value for engineering physiologically relevant infection models.
传统的病毒感染机制研究主要依赖二维（2D）细胞培养模型，这种模型缺乏天然组织的结构组织和生理相关性。这些系统常常无法捕捉到诸如空间细胞间相互作用、组织特异性异质性以及控制体内病毒-宿主动态的微环境复杂性等关键特征。为了解决这些局限性，我们建立了一个整合平台，结合二维和三维（3D）模型的优势，以研究前列腺六跨膜上皮抗原3（STEAP3，一种膜铁还原酶）在调节人类肠道上皮细胞病毒感染中的作用。二维系统能够对STEAP3依赖性病毒进入过程进行高分辨率的机制研究，而患者来源的三维结肠类器官模型重现了肠道组织的结构和细胞复杂性，允许对感染模式进行空间分辨评估。使用这种综合方法，我们发现敲低STEAP3显著增加了病毒进入和感染，特别是在肠上皮细胞和肠内分泌细胞中。为了进一步模拟人体的生理条件，我们开发了一种血管化的芯片类器官模型，在该模型中，STEAP3缺失后在血管腔内观察到病毒信号增加，这表明STEAP3在限制病毒传播中具有保护作用。为了高效、多重地筛选抗病毒机制，我们还制造了一种为类器官培养量身定制的3D打印27孔芯片。通过利用二维和三维系统的互补优势，本研究证明了整合仿生建模平台在研究抗病毒防御机制方面的强大作用，并强调了它们在构建生理相关感染模型方面的价值。
