Creating a 3D dynamic in vitro model of the tumor microenvironment to characteriz

描述（由申请人提供）：炎症被广泛认为在恶性肿瘤的发生和发展中起着核心作用。循环巨噬细胞通过分泌集落刺激因子-1 （CSF-1）和C-C趋化因子配体2 （CCL- 2）等可溶性因子被肿瘤细胞募集。通过这些机制，巨噬细胞被迫获得营养表型，同时释放可溶因子，促进血管生成、肿瘤细胞侵袭和体内浸润。特别令人感兴趣的是tam衍生的肿瘤坏死因子- α (TNF-)，它被认为会导致肿瘤细胞中细胞粘附分子（如E-cadherin）的下调，增加细胞运动性和进入循环。了解这种关系是设计抗转移疗法的关键。然而，该领域报道的大部分数据都是在异种移植模型和/或2D培养中进行的；这是有限的可控变量的数量，外推到人类肿瘤生理学，不适合高通量设计。在这个拟议的研究培训奖学金中，目标是创建一个最佳的体外模型来研究肿瘤微环境。第一个目标是通过一系列经典的共培养实验，更好地了解肿瘤细胞、内皮细胞和tam在二维环境中的行为。该目标的结果将产生相关的设计参数，这些参数将在本项目的第二个目标中实现，该目标将微流体原理与组织工程相结合，创建具有灌注的人体毛细血管的肿瘤微环境的3D组织模型。该模型将能够复制体内肿瘤微环境的生理学；从而提供了相关的生理结果。最重要的是，用灌注的人毛细血管床建立体外3D转移模型可以显著提高高通量抗转移药物的筛选。

DESCRIPTION (provided by applicant): Inflammation is widely recognized to play a central role in the initiation and progression of tumor malignancy. Circulating macrophages are recruited by tumor cells through the secretion of soluble factors such as colony stimulating factor-1 (CSF-1) and C-C chemokine ligand 2 (CCL- 2). Through these mechanisms the macrophages are coerced to acquire a trophic phenotype accompanied by the release of soluble factors which promote angiogenesis, tumor cell invasion, and intravasation. Of particular interest is TAM-derived tumor necrosis factor-alpha (TNF-), which is believed to cause the downregulation of cell adhesion molecules (e.g. E-cadherin) in tumor cells, increasing cell motility and entry into the circulation. Understanding of such relationship is key for the design of anti-metastatic therapeutics. However, much of the data reported in this field has been performed in xenograft models and/or 2D cultures; which are limited by the number of controllable variables, extrapolation to human tumor physiology, and not amenable for a high-throughput design. In this proposed research training fellowship, the goal is to create an optimal in vitro model to study the tumor microenvironment. The first aim seeks to gain a better understanding of the behavior of tumor cells, endothelial cells, and TAMs in a 2D environment through an array of classical co-culture experiments. Results of this aim will generate relevant design parameters which will be implemented in the second aim of this project, which combines principles of microfluidics with tissue engineering to create a 3D tissue model of the tumor microenvironment with perfused human capillaries. This model will be able to replicate the physiology of the in vivo tumor microenvironment; thus providing relevant physiological results. Most importantly, the impact of creating an in vitro 3D metastasis model with perfused human capillary bed could significantly enhance high-throughput anti-metastatic drug screening.