A vascularized 3D bioengineered lung tumor model for anticancer drug screening

Abstract The limited clinical translation of preclinical anticancer drug efficacy underscores the urgent need for advanced models that faithfully replicate tumor pathophysiology. While three-dimensional (3D) tumor cultures improve the fidelity of microenvironmental modeling, most existing systems lack vascularization-a critical element influencing tumor progression and treatment resistance. In this study, we established a vascularized 3D lung cancer model by co-seeding decellularized lung scaffold with human embryonic stem cell-derived endothelial cells, pericytes, and A549 adenocarcinoma cells. This tri-culture system successfully formed a hierarchical vascular network and recapitulated key features of the tumor microenvironment, including hypoxia-driven lysyl oxidase (LOX) overexpression, and integrin-mediated fibronectin-rich desmoplastic niches accumulation. Compared to traditional 2D cultures, this 3D bioengineered platform demonstrated excellent cell compatibility and architectural complexity, supporting enhanced cell migration and MUC5AC hypersecretion. Importantly, cancer cells cultured in this 3D vascularized system exhibited reduced chemosensitivity relative to monolayer cultures. Moreover, patient-derived lung cancer organoids were integrated into the pre-vascularized 3D compartment for individualized drug response testing. Mechanically, hypoxia-activated HIF-1α/LOX signaling promoted ITGA5/FN1-dependent extracellular matrix remodeling, contributing to a chemoprotective niche. This vascularized 3D lung cancer model offers a physiologically relevant and translationally valuable platform for investigating non-small cell lung cancer progression and optimizing patient-specific drug screening.