(PQD5) Development of in Vitro Vascularized Microtumors for Drug Screening

描述(申请人提供)：由于目前预测新药候选药物在人体的疗效或毒性的方法往往不准确，我们能否开发新的方法来测试潜在的治疗药物，从而产生更好的反应预测？挑衅性的问题需要大胆的回答。在这里，我们提出了一种全新的生物工程方法来筛选药物，该方法使用排列的人体3D微肿瘤，并由灌流的毛细血管支持。肿瘤细胞生长在一个充满复杂的细胞外基质和基质细胞的小房间里，并由连接到人造动静脉系统的新形成的毛细血管网络滋养。数百个腔室可以布置在一个芯片上，每个腔室都有自己的血管供应，从而允许开发高通量筛查(HTS)平台。肿瘤株可用于药物发现，或患者特定的肿瘤细胞用于个性化筛查。该系统结合了组织培养的定义、快速、成本效益和可重复性的优点，以及小鼠模型的优点-三维、多种细胞类型相互作用、复杂的药效学以及依赖血管系统生存和药物输送。此外，慢病毒介导的红色、黄色和绿色荧光蛋白在肿瘤细胞、内皮细胞和间质细胞中的表达将允许随着时间的推移重复、快速和定量地评估针对每种细胞类型的药物毒性/疗效。重要的是，这项提议背后的关键技术已经通过了概念验证阶段--我们现在寻求将组件优化和协调到一个工作的HTS平台中，该平台将更准确地预测药物在人体内的作用。

DESCRIPTION (provided by applicant): Since current methods to predict the efficacy or toxicity of new drug candidates in humans are often inaccurate, can we develop new methods to test potential therapeutic agents that yield better predictions of response? Provocative questions demand bold answers. Here we propose a radically new bioengineering approach to drug screening that employs arrayed human 3D microtumors, supported by perfused capillaries. Tumor cells grow within a small chamber filled with complex ECM and stromal cells, and are nourished by a network of newly-formed capillaries that are connected to an artificial arteriovenous system. Hundreds of chambers can be arranged on a single chip, each with its own vascular supply, thereby allowing for development of a high- throughput screening (HTS) platform. Tumor lines can be used for drug-discovery, or patient-specific tumor cells for personalized screening. This system combines the advantages of tissue culture - defined, rapid, cost- effective and reproducible - with the advantages of mouse models - 3-dimensional, multiple cell types interacting, complex pharmacodynamics, and dependence on vasculature for survival and drug-delivery. In addition, lentiviral-mediated expression of red, yellow and green fluorescent proteins in the tumor cells, endothelial cells and stromal cells will allow for repeate, rapid and quantitative assessment of drug toxicity/efficacy against each cell type over time. Importantly, the key technologies underlying this proposal have already passed the proof-of-concept stage - we now seek to optimize and harmonize the components into a working HTS platform that will more accurately predict how drugs will act in humans.