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

项目摘要/摘要 90%以上可以通过临床前验证的候选药物最终未能获得批准 临床应用。这些失败的一个主要原因是传统的模型-单层培养细胞 和实验室动物--在概括本地微环境的能力方面是有限的。这很好 展示了3D培养细胞，通常是球体和支架，可以更接近地模拟它们的自然 行为。有机物质和3D生物打印的新兴技术已经显著发展到 提高这些3D平台的复杂性。然而，缺乏来自周围生物的引导 提示，细胞的固有能力可能不足以驱动靶组织的自组织 在成熟和发育过程中发挥作用。动态模拟生理微环境 具有时空准确性，并在体外重建定向的细胞-细胞和细胞-细胞外基质相互作用，在 具体目标1，3D支持矩阵将编织嵌入生物化学和生物力学 暗示。这将通过可编程的3D打印加载细胞的ECM衍生材料与即时墨水配方以及随后由封装的成纤维细胞引导的ECM重塑来实现。特定目标2将 探索产生的化学和机械双梯度是否可以指导多尺度血管形成 印刷组织结构，这是组织工程中的一个主要挑战。血管系统的可灌注性将 被评估。3D肿瘤模型将组装成药物试验床，以研究基质屏障是如何形成的 治疗药物的耐药性。拟议的生物制造具有三项创新 策略，包括1)3D打印ECM衍生矩阵和生物墨水的即时可编程配方 2)细胞直接迁移、细胞外基质重塑和血管生成 生物化学和生物力学双梯度；3)结合直接三维构建组织结构 生物打印和打印后引导重塑，走向4D生物打印。如果成功，肿瘤实质细胞， 多尺度血管系统和ECM屏障将通过可编程的 过程，为多特异性、高通量药物筛选提供三个潜在的治疗靶点

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