基于高通量单细胞蛋白分析芯片的肿瘤细胞迁移研究

Metastasis is the most deadly reason for cancer patients, while cancer cell migration is the initial step. Migration has been found to be largely mediated by secreted proteins in tumor microenvironment, like cytokines, chemokines, etc. Thus, deep understanding of the correlation between migration and secreted proteins will be of great clinical value to inhibit metastasis by minimizing cancer cell migration. Based on our previous results on single cell proteomic analysis technology, here we propose to establish a three dimensional single cell migration analysis microchip, in which micro-patterned cell culture substrate will be incorporated into single cell analysis microchip for co-detection of cell migration trajectories and their corresponding protein secretion profiles from the same single cells. Besides, in vivo tumor microenvironment conditions like hypoxia, coexistence with macrophage cells will also be established on this platform. The correlation between cell migration and secreted proteins can thus be fully investigated, which can be used to minimize cell migration by tailoring corresponding proteins. The successful implementation of the proposed project will lead to a broadly applicable single cell proteomic analysis platform to investigate cancer cell migration, which could potentially become a precision analysis tool for tumor metastasis research.

转移是导致肿瘤患者死亡的最主要原因，而肿瘤细胞迁移是转移的第一步。肿瘤微环境中的细胞因子、化学因子等分泌蛋白已被发现是促进肿瘤细胞迁移的关键因素，深入认识肿瘤细胞迁移与相关蛋白关系将有助于人们第一时间阻止肿瘤转移。拟研项目提出建立肿瘤迁移单细胞蛋白分析芯片以更深入认识肿瘤细胞迁移。项目以申请人在单细胞蛋白分析技术的积累为基础，引入细胞三维培养基底以诱导肿瘤单细胞的迁移，并构建、模拟多种肿瘤体内微环境（如缺氧、巨噬细胞共培养等），继而对肿瘤单细胞迁移情况和相关蛋白分泌进行研究，以期最终实现抑制肿瘤细胞迁移的目标。本项目最终将形成一个具有一定普适性、能够模拟肿瘤细胞迁移研究的单细胞分析平台，为肿瘤转移的精准研究提供技术支持。

本项目致力于发展单细胞分泌分析芯片技术以深入认识肿瘤细胞迁移与分泌因子的关系，以更好的帮助实现抑制肿瘤细胞迁移。在本项目的资助下，课题组先后完成了高通量单细胞外囊泡分泌分析平台的建立、基于单细胞分泌因子多种类检测技术的细胞相互作用分析平台的建立与应用、单细胞蛋白分析的三维培养环境构建及基于台式扫描仪实现低成本单细胞外囊泡分泌分析等研究内容。项目执行期间，已陆续在国际刊物上以通讯作者发表10篇高质量学术论文，其中包括国际著名综合期刊PNAS2篇，在分析化学领域主流期刊Analytical Chemistry上2篇（其中一篇被Analytical Chemistry 官方twitter以“Scientists combine a microchip 3D cell culture platform with single-cell secretion analysis to investigate the effects of tumor microenvironments on macrophage immune response.”为题介绍了该工作）；申请发明专利2项；博士后出站1名，联合培养硕士生5名；作为共同主编出版中文、英文专著一部（《器官芯片》，2019.11，科学出版社）；《Biosensors for single-cell analysis》，2021.11，Elsevier出版社）。