An automated high-throughput tissue model for screening metastatic effectors

描述（由申请人提供）：我们对调节癌症转移的生物学机制的理解很少，部分原因是缺乏相关的高通量测定系统，这些系统允许有效研究所涉及的无数参数。这些参数不仅包括癌细胞，还包括癌细胞在转移级联中的各个步骤中遇到的微环境。例如，为了转移，癌细胞必须离开原发性肿瘤并通过内渗和外渗（进出血管）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侵入/内渗和外渗/定殖。在这里，我们建议开发一个体外高通量模型的内渗和外渗，结合几个基本的结构/功能的关系，在体内发现。此外，我们将验证在体内小鼠模型和原发性和转移性乳腺肿瘤的人体组织样本的微阵列的体外测定。我们将使用新的微制造方法（例如粘性指进）来创建高通量微血管阵列（例如在细胞外基质内创建的内皮内衬管腔）。该分析平台集成了微流体和多细胞类型3D培养，以提供用于识别内渗和外渗效应器的自动化系统-转移级联中的两个关键步骤。最后，我们将应用该检测平台对疑似影响乳腺癌转移的微环境因素进行基于发现的筛选。我们的方法独特地实现了基于筛选的方法，可能会发现治疗干预的新靶点，这些靶点在传统方法中会被错过，传统方法不允许检查癌细胞，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.