用于临床前药物测试的基于 3D 生物打印水凝胶的非小细胞肺癌肿瘤模型

Lung cancer is the most frequently diagnosed malignancy and remains the leading cause of cancer-related deaths worldwide. Non-small cell lung cancer (NSCLC) is the most prevalent subtype, accounting for the majority of these fatalities. Despite numerous approved therapies, the five-year survival rate for NSCLC patients remains poor, largely due to drug resistance and early relapse. Three-dimensional (3D) tumor models that better recapitulate the in vivo conditions of primary NSCLC hold significant potential for advancing both drug discovery and development. In this study, we developed a hydrogel-based, 3D-bioprinted NSCLC model in a tumor-slice format, in which tumor spheroids or organoids of NSCLC and primary CAFs were directly assembled to create a model that mimics the structure and biochemical properties of the tumor microenvironment (TME). Within this bioprinted approach the spatial distribution and compartment ratios can be defined, enhancing reproducibility and enabling a customizable TME reconstruction. A composite bioink consisting of alginate, collagen, and Matrigel was developed to generate a bio-functionalized hydrogel matrix that supports the structural stability and enables the co-culture of different cell types. Several drugs used for first-line treatment of NSCLC were applied to the system to evaluate the treatment efficiency. The printed 3D tumor slices can be cultured for up to 14 days while maintaining their architecture and proliferative capacity. The process is highly reproducible and supports consistent generation of tumor-like constructs suitable for downstream analyses and drug testing. Functional assays including cytotoxicity assays, live imaging, multiplex immunofluorescence staining and 3D imaging were established to assess cell viability, tumor-stromal cell interaction, and treatment response within the printed 3D tumor slices. The culture can also be expanded to include PBMCs to investigate immune cell behavior within the TME, making the model suitable for evaluating immunotherapies. In conclusion, we established a standardized 3D hydrogel-printed model incorporating NSCLC tumor spheroids or organoids and primary CAFs. By reconstructing the TME, this system provides a reproducible and physiologically relevant platform for preclinical drug screening and cancer research. Together with the established analytical methods, this platform serves as a valuable tool for drug discovery and development.