Elitas Meltem, Brower Kara, Lu Yao, Chen Jonathan J, Fan Rong
Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06520, USA.
Lab Chip. 2014 Sep 21;14(18):3582-8. doi: 10.1039/c4lc00676c. Epub 2014 Jul 24.
It is increasingly recognized that infiltrating immune cells contribute to the pathogenesis of a wide range of solid tumors. The paracrine signaling between the tumor and the immune cells alters the functional state of individual tumor cells and, correspondingly, the anticipated response to radiation or chemotherapies, which is of great importance to clinical oncology. Here we present a high-density microchip platform capable of measuring a panel of paracrine signals associated with heterotypic tumor-immune cell interactions in the single-cell, pair-wise manner. The device features a high-content cell capture array of 5000+ sub-nanoliter microchambers for the isolation of single and multi-cell combinations and a multi-plex antibody "barcode" array for multiplexed protein secretion analysis from each microchamber. In this work, we measured a panel of 16 proteins produced from individual glioma cells, individual macrophage cells and varying heterotypic multi-cell combinations of both on the same device. The results show changes of tumor cell functional phenotypes that cannot be explained by an additive effect from isolated single cells and, presumably, can be attributed to the paracrine signaling between macrophage and glioma cells. The protein correlation analysis reveals the key signaling nodes altered by tumor-macrophage communication. This platform enables the novel pair-wise interrogation of heterotypic cell-cell paracrine signaling at the individual cell level with an in-depth analysis of the changing functional phenotypes for different co-culture cell combinations.
人们越来越认识到，浸润性免疫细胞在多种实体瘤的发病机制中起作用。肿瘤与免疫细胞之间的旁分泌信号改变了单个肿瘤细胞的功能状态，相应地也改变了对放疗或化疗的预期反应，这对临床肿瘤学非常重要。在此，我们展示了一种高密度微芯片平台，该平台能够以单细胞、成对的方式测量一组与异型肿瘤-免疫细胞相互作用相关的旁分泌信号。该设备具有一个由5000多个亚纳升微腔组成的高内涵细胞捕获阵列，用于分离单个和多细胞组合，以及一个多重抗体“条形码”阵列，用于对每个微腔进行多重蛋白质分泌分析。在这项工作中，我们在同一设备上测量了单个胶质瘤细胞、单个巨噬细胞以及两者不同异型多细胞组合产生的一组16种蛋白质。结果显示，肿瘤细胞功能表型的变化无法用分离的单个细胞的累加效应来解释，推测这可能归因于巨噬细胞与胶质瘤细胞之间的旁分泌信号。蛋白质相关性分析揭示了肿瘤-巨噬细胞通讯改变的关键信号节点。该平台能够在单个细胞水平上对异型细胞间旁分泌信号进行新型成对研究，并深入分析不同共培养细胞组合中不断变化的功能表型。
