Programmable Construction of 3D in vitro Disease Models Capturing Microenvironment Heterogeneity

项目总结/文摘

PROJECT SUMMARY/ABSTRACT More than 90% of drug candidates that could pass preclinical validations eventually fail to be approved for clinical applications. A main cause of these failures is that conventional models—monolayer cultured cells and lab animals—are limited in the capability to recapitulate the native microenvironments. It is well demonstrated that 3D cultured cells, typically spheroids and scaffolds, can more closely mimic their natural behaviors. The emerging technologies of organoid and 3D bioprinting have been developed to significantly improve the complexity of these 3D platforms. However, lacking the guidance from surrounding biological cues, the intrinsic capability of cells may not be sufficient to drive self-organization for target tissue functions during the maturation and development. To dynamically model physiological microenvironments with spatiotemporal accuracy and re-establish directed cell-cell and cell-ECM interactions in vitro, in specific aim 1, 3D supporting matrices will be weaved with embedded biochemical and biomechanical cues. This will be enabled by programmably 3D printing cell-laden ECM-derived materials with the on-thefly ink formulation and following guided ECM remodeling by encapsulated fibroblasts. Specific aim 2 will explore if generated chemical and mechanical dual-gradients can direct multiscale vascularization of printed tissue constructs, a major challenge in tissue engineering. The perfusability of the vasculature will be evaluated. 3D tumor models will be assembled as a drug testbed to study how stromal barriers shape the resistance on therapeutic agents. Three innovations are featured by the proposed biofabrication strategy, including 1) 3D Print ECM-derived matrices with on-the-fly programmable formulation of bioinks and in situ crosslinking; 2) Direct cell migration, ECM remodeling, and angiogenesis with immobilized biochemical and biomechanical dual-gradients; 3) Construct tissue architectures by combining direct 3D bioprinting and postprint guided remodeling, toward 4D bioprinting. If successful, tumor parenchymal cells, multiscale vasculature and ECM barriers will be integrated within a single 3D model via a programmable process, providing three potential therapeutic targets for multi-specific, high-throughput drug screening