A Superhydrophobic 3D Cell Culture System Reveals the Mechanobiological Role of Cancer-Associated Fibroblasts in Prostate Cancer Metastasis.

Abstract

The ability to model metastatic dissemination under physiologically relevant mechanical conditions, especially of aggregated circulating tumor cells (CTCs), remains a challenge in cancer research. To address this need, this work presents the Advanced Tumor Landscape Analysis System (ATLAS), a rapidly fabricated, 3D-printed superhydrophobic array platform. ATLAS preserves the hierarchical micro- and nanoscale roughness and low-surface-energy interfaces required for stable superhydrophobicity while greatly reducing fabrication time compared to similar technologies, enabling rapid iteration and broad experimental accessibility. Using a superhydrophobic microwell device, heterotypic tumor-stroma clusters were found to exhibit enhanced survival, sustained proliferation, and coordinated activation of STAT3, AKT1, and NFκB signaling under physiological shear conditions that are lethal to single cancer cells. It was determined that shear exposure reprograms cancer-associated fibroblasts to secrete elevated levels of pro-metastatic cytokines, including IL-11 and CXCL12, with signaling effects that persist well beyond the mechanical stimulus. These findings reveal that mechanical conditioning and activated stromal inclusion jointly drive survival-dominant signaling states characteristic of metastatic fitness. Together, ATLAS establishes a materials-enabled framework for resolving how mechanical forces and multicellular organization converge to shape metastatic behavior, offering a powerful preclinical platform for cancer modeling and translational discovery.