Engineered Invasive Human Breast Tumors with Integrated Capillaries and Lymphatics

项目摘要 目前人类癌症的体内和体外模型的复制能力仍然有限 以易于获取和生理相关的形式向侵袭性疾病进展。组织工程 肿瘤可以通过对肿瘤的关键方面进行模块化控制来提供更强大的系统， 微环境，如血管密度或间质压力。这项合作研究旨在开发 并应用新的体外血管化肿瘤工程方法，其中细胞，物理和遗传 可以以高的空间和时间分辨率控制肿瘤的组成及其微环境。 该合作团队由生物材料和组织工程（Tien），定量 发育和肿瘤生物学（纳尔逊）、力学（Ekinci）和临床肿瘤生物学（Radisky）以及 病理学（Nassar）。 我们在过去15年中一直在开发的核心技术是使用 三维（3D）微图案化的细胞外基质水凝胶作为支架， 组织工程化组织。具体来说，拟议的工作将创造微型人类乳房 含有灌注毛细血管和引流毛细血管的肿瘤，为肿瘤细胞逃逸提供途径 并能够捕获这些细胞用于下游表达谱分析。间质应力和生物化学 将分别通过非侵入性成像和间质液的重复取样来分析组合物，以 提供与肿瘤细胞行为相关的纵向数据。这项工作还将创造血管化的 可以接受人乳腺肿瘤活检物作为体外“患者来源的异种移植物”的胶原基质， 发现有利于肿瘤侵袭和逃逸的候选突变;然后对这些突变进行测试 在使用工程乳腺肿瘤的假设驱动的分析中。更广泛地说，这项工作将传播 这些微型组织工程技术的癌症研究实验室，以适应其他类型 癌症和肿瘤细胞的行为。

PROJECT SUMMARY Current in vivo and in vitro models of human cancer remain limited in their ability to replicate progression to invasive disease in an easily accessible and physiologically relevant format. Tissue-engineered tumors may provide a more powerful system by enabling modular control over key aspects of a tumor and its microenvironment, such as vascular density or interstitial pressure. This collaborative study seeks to develop and apply new methods of engineering vascularized tumors in vitro, in which the cellular, physical, and genetic composition of the tumor and its microenvironment can be controlled with high spatial and temporal resolution. The collaborative team consists of experts in biomaterials and tissue engineering (Tien), quantitative developmental and tumor biology (Nelson), mechanics (Ekinci), and clinical tumor biology (Radisky) and pathology (Nassar). The core enabling technology, which we have been developing over the past fifteen years, is the use of three-dimensional (3D) micropatterned extracellular matrix hydrogels as scaffolds for directing the 3D organization of engineered tissues. Specifically, the proposed work will create microscale human breast tumors that contain perfused capillaries and draining lymphatics, which provide routes for tumor cell escape and enable the capture of those cells for downstream expression profiling. Interstitial stresses and biochemical composition will be analyzed by non-invasive imaging and repeated sampling of interstitial fluid, respectively, to provide longitudinal data for correlation with tumor cell behavior. This work will also create vascularized collagenous stroma that can accept human breast tumor biopsies as in vitro "patient-derived xenografts", for the discovery of candidate mutations that favor tumor invasion and escape; these mutations will then be tested in hypothesis-driven analyses using the engineered breast tumors. More broadly, this work will disseminate these microscale tissue engineering technologies to cancer research laboratories for adaptation to other types of cancers and tumor cell behaviors.