Department of Bioengineering, University of Washington, Seattle, WA 98105, USA.
Department of Surgery, University of Washington, Seattle, WA 98105, USA.
Sci Adv. 2024 Sep 6;10(36):eadn5875. doi: 10.1126/sciadv.adn5875.
Functional assays on intact tumor biopsies can complement genomics-based approaches for precision oncology, drug testing, and organs-on-chips cancer disease models by capturing key therapeutic response determinants, such as tissue architecture, tumor heterogeneity, and the tumor microenvironment. Most of these assays rely on fluorescent labeling, a semiquantitative method best suited for single-time-point assays or labor-intensive immunostaining analysis. Here, we report integrated aptamer electrochemical sensors for on-chip, real-time monitoring of cytochrome C, a cell death indicator, from intact microdissected tissues with high affinity and specificity. The platform features a multi-well sensor layout and a multiplexed electronic setup. The aptasensors measure increases in cytochrome C in the supernatant of mouse or human microdissected tumors after exposure to various drug treatments. Because of the sensor's high affinity, it primarily tracks rising concentrations of cytochrome C, capturing dynamic changes during apoptosis. This approach could help develop more advanced cancer disease models and apply to other complex in vitro disease models, such as organs-on-chips and organoids.
基于功能的完整肿瘤活检分析，可以通过捕获关键的治疗反应决定因素，如组织架构、肿瘤异质性和肿瘤微环境，来补充基于基因组的精准肿瘤学、药物测试和器官芯片癌症疾病模型。这些分析方法中的大多数都依赖于荧光标记，这是一种半定量方法，最适合单次检测或需要大量免疫染色分析的实验。在这里，我们报告了一种集成的适体电化学传感器，用于在芯片上实时监测细胞色素 C，这是一种细胞死亡的指标，其来源于高亲和力和特异性的完整微切割组织。该平台具有多井传感器布局和多路复用电子设置。在将各种药物处理应用于小鼠或人类微切割肿瘤的上清液后，适体传感器可以测量细胞色素 C 的增加。由于传感器的高亲和力，它主要跟踪细胞色素 C 浓度的升高，捕获细胞凋亡过程中的动态变化。这种方法可以帮助开发更先进的癌症疾病模型，并应用于其他复杂的体外疾病模型，如器官芯片和类器官。
