Microfluidic platform for tumor cell invasion

项目摘要 癌症转移占所有癌症死亡的90%以上。癌症转移级联中的一个限制步骤是 肿瘤细胞向血管壁迁移，与血管壁相互作用， 通过血液循环扩散到继发性肿瘤部位。生物物理力，包括间质力和 壁内流动在调节粘附分子、空间细胞因子 分布和组织结构;所有这些都有助于肿瘤细胞在3D生物基质内的侵袭。尽管 生物物理力在肿瘤细胞跨内皮迁移（TEM）中的临床重要性和作用很差， 明白这部分是由于缺乏能够跟踪肿瘤细胞迁移的体外工具 事件在真实的时间，并与良好控制的生物流。目前的动物细胞侵袭试验，Boyden 腔室，是有限的，因为它是难以重建复杂的肿瘤微环境。此外，结果 是基于两个时间终点的人群。活体成像已经大大提高了我们对 关于肿瘤微环境和TEM之间的相互作用，在生理上现实的设置。然而，在这方面， 很难剖析单个环境线索对TEM过程的贡献。的目标 建议的研究是开发一个生理上现实的微流体模型， 用于研究肿瘤细胞TEM过程的肿瘤微环境;以及鉴定 促进TEM。为了实现这些目标，我们将开发一个器官型微流体模型的真实的时间 在良好控制的微环境下对肿瘤细胞TEM事件进行成像。我们将利用 球体和细胞流事件来引导TEM成像位点。使用微流体模型，我们将 在良好的控制条件下，探索单个肿瘤细胞特性与TEM活性的关系， 间质和壁内血流。PI实验室和其他人以前的工作表明， 流动严格地调节肿瘤细胞在3D生物基质内的迁移。在这里，我们假设肿瘤细胞 TEM能力与细胞的微环境（包括流体流动）密切相关。拟议 该项目是创新的，因为它代表了第一代器官型微流体平台， 包括间质流和壁内流，将当前的微流体肿瘤模型推向 生理现实的方向。在此吸取的经验教训将最终导致重要的知识， 开发新的癌症诊断或/和治疗策略。该平台可以随时扩展 用于TEM重要的其它生物系统，包括免疫细胞运输。

Project Summary Cancer metastasis accounts for over 90% of all cancer deaths. A limiting step in cancer metastatic cascade is for tumor cells to migrate towards, interact with and squeeze through the blood vessel wall before disseminating to secondary tumor sites via the blood circulation. Biophysical forces, including interstitial and intramural flows, have shown to play critical roles in regulating adhesion molecules, spatial cytokine distributions and tissue architecture; all of which contribute to tumor cell invasion within 3D biomatrix. Despite the clinical importance, roles of biophysical forces in tumor cell transendothelial migration (TEM) are poorly understood. This is in part due to the lack of in vitro tools that are able to follow tumor cell transmigration events in real time, and with well controlled biological flows. Current animal cell invasion assay, the Boyden Chamber, is limited because it is difficult to recreate complex tumor microenvironment. In addition, the results are population based at two time end points. Intravital imaging has advanced significantly our understanding about the interplays between tumor microenvironment and TEM in a physiologically realistic setting. However, it is difficult to dissect the contribution of individual environmental cues to TEM processes. The goals of the proposed research are to develop a physiologically realistic microfluidic model with well controlled tumor microenvironment for studies of tumor cell TEM processes; and to identify tumor microenvironment that promotes TEM. To achieve these goals, we will develop an organotypic microfluidic model for real time imaging of tumor cell TEM events under well controlled micro-environment. We will use the location of spheroid and cell streaming event to guide TEM imaging sites. Using the microfluidic model, we will explore the relations between single tumor cell properties and TEM activities under well controlled interstitial and intramural flows. Previous work from the PI’s lab and others have indicated that interstitial flows critically regulate tumor cell migration within 3D biomatrix. Here, we hypothesize that tumor cells’ TEM capabilities are closed correlated with cells’ microenvironment including fluid flows. The proposed project is innovative because it represents the first generation of organotypic microfluidic platform that includes both interstitial and intramural flows, moving the current microfluidic tumor model towards a physiologically realistic direction. Lessons learned here will eventually lead to knowledge important for developing novel diagnostic or/and treatment strategies for cancer. This platform can be readily extended for use in other biological systems where TEMs are important including immune cell trafficking.