Magnetically patterned co-cultures for cancer studies

描述(申请人提供)：癌细胞与其周围微环境之间的粘附性相互作用在调节实体肿瘤的生长、发展和侵袭方面起着关键作用。癌细胞和组织基质细胞之间的直接黏附接触被认为有助于癌症的存活、增殖和迁移。然而，这种异型细胞-细胞相互作用对癌细胞功能调节的贡献尚不清楚，因为这些相互作用很难在实验上操作。该项目将结合两名研究人员的专业知识，开发一种新的系统，在微图案化培养中并列两种细胞类型，以便能够以可控的方式大量引入高保真的异型细胞-细胞相互作用。该方法包括使用磁性纳米线、微制造的磁性陷阱阵列，并设计这些陷阱与细胞的粘附性。该项目的长期目标包括开发一种方法来识别调节癌细胞功能的异型细胞-细胞相互作用。作为概念验证，这一新方法将用于识别与一种间质细胞--内皮细胞接触时促进增殖的癌细胞系。具体目标1将是优化和表征影响捕获在经历异型细胞-细胞相互作用的阵列上的细胞的产量和总数的参数。具体目标2将确定直接接触内皮细胞对模型癌细胞系增殖的影响。具体目标3将通过生化分析演示该系统的使用，并为调控癌症增殖的异型癌症-间质细胞-细胞对相互作用的机制研究奠定基础。这项研究将应用磁性微米和纳米技术的最新进展，开发一种研究癌症生物学的新方法。即将开发的设备将使研究癌细胞与周围健康组织的相互作用成为可能，这是以前无法实现的。对这一新方法将带来的更多了解将有可能为控制癌症的生长和扩散的努力提供重要的投入。

DESCRIPTION (provided by applicant): Adhesive interactions between cancer cells and their surrounding microenvironment play a critical role in regulating the growth, development, and invasion of solid tumors. Direct adhesive contact between cancer cells and tissue stromal cells are thought to contribute to the survival, proliferation, and migration of cancers. However, the contribution of such heterotypic cell-cell interactions to the regulation of cancer cell function remains unclear because these interactions have been difficult to manipulate experimentally. This project will combine the expertise of two investigators to develop a novel system to juxtapose two cell types in micropatterned culture such that high fidelity heterotypic cell-cell interactions can be introduced in a controllable fashion in large numbers. The approach involves the use of magnetic nanowires, microfabricated arrays of magnetic traps, and engineering the adhesiveness of these traps to cells. The long-term goals of this project involve the development of a method to identify heterotypic cell-cell interactions that regulate the function of cancer cells. As a proof-of-concept, this novel approach will be used to identify cancer cell lines that increase proliferation in contact with one stromal cell, the endothelial cell. Specific Aim 1 will be to optimize and characterize the parameters that affect the yield and total number of cells trapped on the arrays that experience heterotypic cell-cell interactions. Specific Aim 2 will be to determine the effect of direct contact with endothelial cells on the proliferation of a model carcinoma cell line. Specific Aim 3 will be to demonstrate the use of this system with biochemical assays and to lay the groundwork for mechanistic studies of heterotypic cancer-stromal cell-cell pair interactions that modulate cancer proliferation. This research will apply recent advances in magnetic micro- and nanotechnology to develop a new approach to study the biology of cancer. The devices to be developed will enable studies of the interactions of cancer cells with surrounding healthy tissue that have not been possible previously. The increased understanding that this new approach will bring will have the potential to provide important input into efforts to control the growth and spread of cancer.