Mechanosensitive interactions of tumoroids with an engineered environment promote cell proliferation and enhance drug response detection

Despite remarkable advances in reproducing ever more sophisticated ex-vivo models, foundational challenges remain in standardizing and scaling up organoid culture. Here, we introduce a novel technology for high-throughput derivation and high-content analysis of tumoroids. This technology relied on microengineered tumoroid niches fabricated from PEG acrylate polymers by maskless photolithography to create a mechanically tunable microenvironment that is scalable and compatible with standard automated lab pipelines. We validated our innovative platform on patient-derived urothelial tumoroids and evaluated its use in high-end imaging approaches, including holotomography. Our results reveal that tumoroids dynamically interact and deform their synthetic environment, unravelling a mechanosensitive behavior of tumoroids depending on the features of their synthetic niche. Changing mechanical properties of microcavities modulated cell cycle dynamics and metabolism, as it affected their response to drugs, which highlights the critical role of mechanical stimuli in tumoroid development and future drug screening. Soft niches condition : M_N1/ M_N2/ M_N3/ M_N4 Stiff niches condition : S_N1/ S_N2/ S_N3/ S_N4 2D condition : 2D_N1/ 2D_N2/ 2D_N3/ 2D_N4