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

描述(申请人提供)：本申请针对PQ#24：考虑到研究转移的困难，我们能否开发新的方法，例如工程组织移植，来研究肿瘤扩散的生物学？转移过程中的许多步骤，特别是侵袭、内渗和外渗，发生在局部组织微环境和血管系统之间的交界处或附近。因此，开发一种结合细胞外基质和血管的平台是解开引导转移发生的事件的关键。开发这样一个平台的主要挑战是这个界面的复杂性。为了应对这一挑战，我们提出了一种微流控平台，它结合了人工细胞外基质和血管。我们的目标是创造一个平台：(1)在生理相关的几何形状中概括细胞外基质和血管之间界面的相关物理和生物特征，(2)允许控制物理化学和生物属性，以便可以系统地和可重复地执行实验，并允许独立调整变量，以及(3)足够坚固，可以很容易地将制造转移到其他实验室。在……里面 初步数据显示，我们已经制作了一个功能平台，并证明了利用该平台研究转移的可行性。在这项研究中，我们建议为该平台的完善和进一步开发奠定基础，使对转移的理解能够取得进展。人造细胞外基质/血管平台允许在生理相关的几何学中研究侵袭、内渗和外渗。为了研究侵袭和血管内侵入，在人工血管附近的细胞外基质中创建了一个空洞。癌细胞的增殖、脱离和迁移到血管，然后进入血管，可以实时成像。为了研究外渗，将癌细胞加入流经血管的灌流介质中。根据血管的大小，可以通过粘连或闭塞来发生停搏。在初步数据中，我们进行了使用血管内皮细胞和血管内皮细胞形成灌流人工血管的原理验证演示 为了研究肿瘤的侵袭和血管内渗而结合的肿瘤。该项目的总体目标是开发一个工程化的ECM/血管平台，用于系统研究转移级联中的关键步骤。在这些结果的基础上，我们将优化工程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，并开发用于研究转移研究的工程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).