An engineered platform for the study of metastasis (PQ #24)

描述（由申请人提供）：本申请涉及PQ #24：鉴于研究转移的难度，我们能否开发新方法（如工程组织移植物）来研究肿瘤扩散的生物学？转移过程中的许多步骤，特别是侵入、内渗和外渗，发生在局部组织微环境和血管系统之间的界面处或附近。因此，结合细胞外基质和血管的平台的开发是解开引导转移发展的事件的关键。开发这样一个平台的主要挑战是这个接口的复杂性。为了应对这一挑战，我们提出了一种微流体平台，它结合了人工细胞外基质和血管。我们的目标是建立一个平台，（1）以生理学相关的几何形状概括细胞外基质和血管之间的界面的相关物理和生物学特性，（2）允许控制物理化学和生物学特性，使得实验可以系统地和可再现地进行，并且允许独立地调节变量，以及（3）足够坚固，使得制造可以容易地转移到其他实验室。在 初步数据我们已经证明了功能平台的制造和使用该平台研究转移的可行性。在这项研究中，我们建议为平台的完善和进一步发展奠定基础，以促进对转移的理解。人工细胞外基质/血管平台允许在生理学相关几何结构中研究侵入、内渗和外渗。为了研究侵入和内渗，在人造血管附近的细胞外基质中创建空腔。癌细胞向血管的转移、脱离和迁移，随后向血管内渗透，可以真实的实时成像。为了研究外渗，将癌细胞添加到流过血管的灌注介质中。根据血管大小，可通过粘连或闭塞发生停搏。在初步的数据中，我们已经进行了使用血管内皮细胞形成灌注人工血管的原理证明， 为了研究肿瘤的侵袭和内渗而将肿瘤合并。该项目的总体目标是开发一个工程ECM/血管平台，用于转移级联中关键步骤的系统研究。在这些结果的基础上，我们将优化工程化ECM/血管平台（目标1），研究侵袭和内渗（目标2a）和外渗（目标2b）的动力学，并开发用于转移研究的工程化ECM/血管平台的翻译模块（目标3）。

DESCRIPTION (provided by applicant): This application addresses PQ #24: Given the difficulty of studying metastasis, can we develop new approaches, such as engineered tissue grafts, to investigate the biology of tumor spread? Many of the steps in the metastatic process, specifically invasion, intravasation, and extravasation, take place at or near the interface between the local tissue microenvironment and the vascular system. Therefore the development of a platform that combines both extracellular matrix and a vessel is key to unraveling the events that guide the development of metastasis. The major challenge in developing such a platform is the complexity of this interface. To address this challenge we propose a microfluidic platform that incorporates both artificial extra cellular matrix and a vessel. Our objective is to produce a platform that: (1) recapitulates the relevant physical and biological characteristics of the interface between extracellular matrix and a vessel in a physiologically relevant geometry, (2) allows control over physicochemical and biological properties such that experiments can be performed systematically and reproducibly, and allowing variables to be adjusted independently, and (3) is sufficiently robust that fabrication can be readily translated to other laboratories. In preliminary data we have demonstrated fabrication of a functional platform and the feasibility of using the platform to study metastasis. In this research, we propose to lay the foundations for the refinement and further development of the platform to enable advances in the understanding of metastasis. The artificial extra cellular matrix/vessel platform allows study of invasion, intravasation and extravasation in a physiologically relevant geometry. To study invasion and intravasation a cavity is created in the extra cellular matrix near the artificial vessel. Prolifertion, detachment, and migration of cancer cells to the vessel, followed by intravasation into the vessel, can be imaged in real time. To study extravasation, cancer cells are added to the perfusion media flowing through the vessel. Depending on the vessel size, arrest can occur by adhesion or occlusion. In preliminary data, we have performed a proof-of-principle demonstration of the formation of a perfused artificial vessel using vascular endothelial cells and the incorporation of a tumor for the study of invasion and intravasation. The overall goal of this project is to develop an engineered ECM/vessel platform for the systematic study of key steps in the metastatic cascade. Building on these results we will optimize the engineered ECM/vessel platform (Aim 1), study the dynamics of invasion and intravasation (Aim 2a) and extravasation (Aim 2b), and develop modules for the translation of the engineered ECM/vessel platform for the study of metastasis (Aim 3).