Quantitative analyses of tumor cell extravasation

 描述（由申请人提供）：由于需要复制体内情况的模型，但又易于严格控制并有助于高分辨率、延时成像和细胞行为的定量分析，因此对导致转移性癌症的基本机制的理解受到阻碍。在过去的几年里，我们已经开发出能够模拟转移的许多步骤的微流体系统，包括肿瘤细胞侵袭、内渗、微循环中的捕获或粘附到血管壁， 外渗到周围的细胞外基质中。这一先前的工作揭示了迁移肿瘤细胞和内皮之间的相互作用，特异性粘附分子的作用，以及细胞特别是细胞核的变形，在迁移过程中的经验。这项拟议研究的目的是采用这些最近开发的测定与新的测量方法相结合，以询问细胞力学的变化过程中的外渗，并了解细胞和细胞-基质力的相互作用的性质。我们还旨在研究核变形，染色质结构的变化以及由此产生的转录组变化，这可能对外渗细胞形成新肿瘤的后续能力具有重要意义。与这些实验密切配合，将开发计算模型来模拟肿瘤细胞/内皮细胞相互作用，核变形以及由此产生的基因表达变化。我们预计，这些研究将提供新的见解，并可能提高我们识别和筛选新疗法的能力，以抑制疾病转移扩散的趋势。

 DESCRIPTION (provided by applicant): Understanding of the fundamental mechanisms leading to metastatic cancer has been hampered by the need for models that replicate the in vivo situation, yet are amenable to tight control and facilitate high-resolution, time-lapse imagin and quantitative analysis of cell behavior. Over the past several years, we have developed microfluidic systems that are capable of simulating many steps of metastasis including tumor cell invasion, intravasation, trapping in the microcirculation or adhesion to the vessel walls, and extravasation into surrounding extracellular matrix. This prior work has shed new light on the interactions between a transmigrating tumor cell and the endothelium, the role of specific adhesion molecules, and the deformations of the cell and especially the cell nucleus, experience during the transmigration process. The object of this proposed study is to employ these recently developed assays in combination with new measurement methods to interrogate the changes in cell mechanics during the process of extravasation, and understand the nature of the cell-cell and cell-matrix force interactions. We also aim to investigate the nuclear deformations, changes in chromatin structure and the resulting changes in the transcriptome, which could have important implications for the subsequent ability of extravasated cells to form a new tumor. In close coordination with these experiments, computational models will be developed to simulate tumor cell / endothelial cell interactions, nuclear deformation, and the resulting changes in gene expression. We anticipate that these studies will provide new insights, and potentially enhance our ability to identify and screen for new therapies to inhibit the tendency for metastatic spread of disease.