An automated high-throughput tissue model for screening metastatic effectors

描述(由申请人提供)：我们对调控癌症转移的生物学机制的了解很少，部分原因是缺乏相关的高通量分析系统，允许对涉及的无数参数进行有效研究。这些参数不仅包括癌细胞，还包括癌细胞在转移级联的不同阶段遇到的微环境。例如，为了转移，癌细胞必须离开原发肿瘤，通过血管内和外渗(进入和离开血管)在循环系统中导航。这发生在由不同相互作用的细胞类型组成的复杂环境中，这些细胞类型可能影响和调节入侵/内渗和外渗/定植。在这里，我们建议建立一个体外高通量的血管内和外渗模型，该模型结合了体内发现的几种基本结构/功能关系。此外，我们将用活体小鼠模型和来自原发和转移性乳腺肿瘤的人类组织样本微阵列来验证体外试验。我们将使用新的微制造方法(例如，粘性指进)来创建高通量的微血管阵列(例如，在细胞外基质中创建的内皮衬里管腔)。该检测平台集成了微流体和多种细胞类型的3D培养，提供了一个自动系统来识别血管内和外渗的影响因素-转移级联中的两个关键步骤。最后，我们将应用该分析平台对可能影响乳腺癌转移的微环境因素进行基于发现的筛选。由我们的方法独一无二地实现的基于筛查的方法可能会发现治疗干预的新靶点，而传统方法无法检查癌细胞、基质细胞和细胞外基质之间的复杂相互作用。

DESCRIPTION (provided by applicant): Our understanding of the biological mechanisms that regulate cancer metastasis are poorly understood, in part, because of the lack of relevant and high-throughput assay systems that allow for the efficient study of the myriad of parameters involved. These parameters include not just the cancer cells, but the microenvironment the cancer cells encounter at various steps in the metastatic cascade. For example, in order to metastasize, cancer cells must leave the primary tumor and navigate the circulatory system via intravasation and extravasation (into and out of blood vessels). This occurs amidst a complex environment consisting of different interacting cell types that likely effect and regulate invasion/intravasation and extravasation/colonization. Here we propose to develop an in vitro high throughput model of intravasation and extravasation that incorporates several essential structure/function relationships found in vivo. Further, we will validate the in vitro assay with a in vivo mouse model and a microarray of human tissue samples from primary and metastatic mammary tumors. We will use novel micro fabrication methods (e.g. viscous fingering) to create high-throughput arrays of micro vessels (e.g. endothelial lined lumens created within extracellular matrix). The assay platform integrates microfluidics and multiple cell type 3D culture to provide an automated system for identifying the effectors of intravasation and extravasation - two key steps in the metastatic cascade. Finally, we will apply the assay platform to perform a discovery-based screen of the microenvironmental factors suspected to influence breast cancer metastasis. The screening-based approach uniquely enabled by our approach will likely discover new targets for therapeutic intervention that would have been missed in traditional approaches which don't allow an examination of the complex interactions between cancer cells, stromal cells and the extracellular matrix.