Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853-0001, United States.
Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge CB3 0AS, United Kingdom.
ACS Biomater Sci Eng. 2021 Dec 13;7(12):5585-5597. doi: 10.1021/acsbiomaterials.1c01056. Epub 2021 Nov 21.
Cancer-derived exosomes (cEXOs) facilitate transfer of information between tumor and human primary stromal cells, favoring cancer progression. Although the mechanisms used during this information exchange are still not completely understood, it is known that binding is the initial contact established between cEXOs and cells. Hence, studying binding and finding strategies to block it are of great therapeutic value. However, such studies are challenging for a variety of reasons, including the need for human primary cell culture, the difficulty in decoupling and isolating binding from internalization and cargo delivery, and the lack of techniques to detect these specific interactions. In this work, we created a supported biomimetic stem cell membrane incorporating membrane components from human primary adipose-derived stem cells (ADSCs). We formed the supported membrane on glass and on multielectrode arrays to offer the dual option of optical or electrical detection of cEXO binding to the membrane surface. Using our platform, we show that cEXOs bind to the stem cell membrane and that binding is blocked when an antibody to integrin β1, a component of ADSC surface, is exposed to the membrane surface prior to cEXOs. To test the biological outcome of blocking this interaction, we first confirm that adding cEXOs to cultured ADSCs leads to the upregulation of vascular endothelial growth factor, a measure of proangiogenic activity. Next, when ADSCs are first blocked with anti-integrin β1 and then exposed to cEXOs, the upregulation of proangiogenic activity and cell proliferation are significantly reduced. This biomimetic membrane platform is the first cell-free label-free in vitro platform for the recapitulation and study of cEXO binding to human primary stem cells with potential for therapeutic molecule screening as it is compatible with scale-up and multiplexing.
癌症衍生的外泌体 (cEXOs) 促进肿瘤与人体原代基质细胞之间的信息传递，有利于癌症的进展。虽然在此信息交换过程中使用的机制尚不完全清楚，但已知结合是 cEXOs 与细胞之间建立的初始接触。因此，研究结合并寻找阻断它的策略具有重要的治疗价值。然而，由于各种原因，此类研究具有挑战性，包括需要人类原代细胞培养、难以将结合与内化和货物传递分离、以及缺乏检测这些特定相互作用的技术。在这项工作中，我们创建了一种支持的仿生干细胞膜，其中包含来自人体原代脂肪来源干细胞 (ADSCs) 的膜成分。我们在玻璃和多电极阵列上形成支撑膜，提供光学或电检测 cEXO 与膜表面结合的双重选择。使用我们的平台，我们表明 cEXOs 与干细胞膜结合，并且当在 cEXOs 之前将针对整合素 β1 的抗体暴露于膜表面时，结合被阻断。为了测试阻断这种相互作用的生物学结果，我们首先证实向培养的 ADSCs 添加 cEXOs 会导致血管内皮生长因子的上调，这是促血管生成活性的衡量标准。接下来，当首先用抗整合素 β1 阻断 ADSCs 然后暴露于 cEXOs 时，促血管生成活性和细胞增殖的上调显著降低。这种仿生膜平台是第一个无细胞无标记的体外平台，用于模拟 cEXO 与人体原代干细胞的结合，并具有治疗分子筛选的潜力，因为它与规模化和多重化兼容。
