Biomechanical 3D tumor models on a micro-milled high-throughput force sensor array.

Abstract

The tumor microenvironment plays a critical role in drug resistance, with extracellular matrix mechanics, cell-cell crosstalk, and transport barriers contributing to poor therapeutic outcomes. Traditional two-dimensional (2D) cultures fail to capture these features, and drug efficacy in 2D often does not translate to three-dimensional (3D) models or<i>in vivo</i>tumors. Here, we present a 3D tumor model integrated with a high-throughput biomechanical sensor array that enables simultaneous measurement of cellular forces and matrix remodeling. The platform, fabricated using a scalable and cost-effective micro-milling approach, supports the parallel generation of multiple tumor constructs within a single dish. To demonstrate feasibility, we formed<i>in vitro</i>tumors using patient-derived pancreatic ductal adenocarcinoma organoids, cancer cells, and stromal fibroblasts. The sensors were then applied to characterize the evolving biophysical properties of these tumors (tissue force and stiffness) and to evaluate responses to chemotherapy drug, Gemcitabine, and the investigational agent, all-trans retinoic acid. Drug responses in 3D tumors were compared with those in 2D cultures. By combining biochemical and biomechanical readouts, this 3D platform provides a more physiologically relevant tumor model and a powerful tool for preclinical drug testing and personalized medicine.