High-throughput drug screening (HTDS) has significantly reduced the time and cost of new drug development. Nonetheless, contact-dependent cell-cell communication (CDCCC) may impact the chemosensitivity of tumour cells. There is a pressing need for low-cost single-cell HTDS platforms, alongside a deep comprehension of the mechanisms by which CDCCC affects drug efficacy, to fully unveil the efficacy of anticancer drugs. In this study, we develop a microfluidic chip for single-cell HTDS and evaluate the molecular mechanisms impacted by CDCCC using quantitative mass spectrometry-based proteomics. The chip achieves high-quality drug mixing and single-cell capture, with single-cell drug screening results on the chip showing consistency with those on the 96-well plates under varying concentration gradients. Through quantitative proteomic analysis, we deduce that the absence of CDCCC in single tumour cells can enhance their chemoresistance potential, but simultaneously subject them to stronger proliferation inhibition. Additionally, pathway enrichment analysis suggests that CDCCC could impact several signalling pathways in tumour single cells that regulate vital biological processes such as tumour proliferation, adhesion, and invasion. These results offer valuable insights into the potential connection between CDCCC and the chemosensitivity of tumour cells. This research paves the way for the development of single-cell HTDC platforms and holds the promise of advancing tumour personalized treatment strategies.

中文翻译：

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基于单细胞高通量药物筛选平台探讨接触依赖性细胞间通讯对化疗敏感性的影响机制

高通量药物筛选（HTDS）显着减少了新药开发的时间和成本。尽管如此，接触依赖性细胞间通讯（CDCCC）可能会影响肿瘤细胞的化疗敏感性。迫切需要低成本的单细胞HTDS平台，同时深入理解CDCCC影响药效的机制，以全面揭示抗癌药物的功效。在这项研究中，我们开发了一种用于单细胞 HTDS 的微流控芯片，并使用基于定量质谱的蛋白质组学评估了 CDCCC 影响的分子机制。该芯片实现了高质量的药物混合和单细胞捕获，芯片上的单细胞药物筛选结果与96孔板上不同浓度梯度下的单细胞药物筛选结果一致。通过定量蛋白质组学分析，我们推断单个肿瘤细胞中CDCCC的缺失可以增强其化疗耐药潜力，但同时使它们受到更强的增殖抑制。此外，通路富集分析表明，CDCCC 可能影响肿瘤单细胞中调节肿瘤增殖、粘附和侵袭等重要生物过程的多个信号通路。这些结果为 CDCCC 与肿瘤细胞的化疗敏感性之间的潜在联系提供了有价值的见解。这项研究为单细胞 HTDC 平台的开发铺平了道路，并有望推进肿瘤个性化治疗策略。