Biosensor National Special Laboratory, Key Laboratory for Biomedical Engineering of Education Ministry, Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, China; State Key Laboratory of Transducer Technology, Chinese Academy of Sciences, Shanghai 200050, China.
Biosensor National Special Laboratory, Key Laboratory for Biomedical Engineering of Education Ministry, Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, China; Key Laboratory of Healthy & Intelligent Kitchen System Integration of Zhejiang Province, No. 218 Binhai 2nd Road, Ningbo 315336, China.
Talanta. 2019 Jul 1;199:131-139. doi: 10.1016/j.talanta.2019.02.021. Epub 2019 Feb 6.
Detection and identification of bitter compounds draw great attention in pharmaceutical and food industry. Several well-known agonists of specific bitter taste receptors have been found to exhibit anti-cancer effects. For example, N-C=S-containing compounds, such as allyl-isothiocyanates, have shown cancer chemo-preventive effects. It is worth noting that human T2R38 receptor is specific for compounds containing N-C=S moiety. Here, a bioinspired cell-based bioelctronic tongue (BioET) is developed for the high-specificity isothiocyanate-induced bitter detection, utilizing human Caco-2 cells as a primary sensing element and interdigitated impedance sensor as a secondary transducer. As an intestinal carcinoma cell line, Caco-2 endogenously expresses human bitter receptor T2R38, and the activation of T2R38 induces the changes of cellular morphology which can be detected by electric cell-substrate impedance sensing (ECIS). After configuration and optimization of parameters including timing of compound administration and cell density, quantitative bitter evaluation models were built for two well-known bitter compounds, phenylthiocarbamide (PTC) and propylthiouracil (PROP). The bitter specific detection of this BioET is inhibited by probenecid and U-73122, and is not elicited by other taste modalities or bitter ligands that do not activate T2R38. Moreover, by combining different computational tools, we designed a ligand-based virtual screening (LBVS) protocol to select ligands that are likely to activate T2R38 receptor. Three computationally predicted agonists of T2R38 were selected using the LBVS protocol, and the BioET presented response to the predicted agonists, validating the capability of the LBVS protocol. This study suggests this unique cell-based BioET paves a general and promising way to specifically detect N-C=S-containing compounds that can be used for pharmaceutical study and drug development.
苦味化合物的检测和识别在医药和食品工业中引起了极大的关注。已经发现几种已知的特定苦味受体激动剂具有抗癌作用。例如，含有 N-C=S 基团的化合物，如丙烯基异硫氰酸酯，具有抗癌化学预防作用。值得注意的是，人类 T2R38 受体是专门针对含有 N-C=S 部分的化合物。在这里，开发了一种基于生物灵感的细胞生物电子舌（BioET），用于高特异性异硫氰酸盐诱导的苦味检测，利用人 Caco-2 细胞作为主要传感元件和叉指阻抗传感器作为二次换能器。作为一种肠癌细胞系，Caco-2 内源性表达人类苦味受体 T2R38，T2R38 的激活会引起细胞形态的变化，这可以通过电细胞-基底阻抗传感（ECIS）检测到。在包括化合物给药时间和细胞密度等参数的配置和优化后，为两种著名的苦味化合物苯硫脲（PTC）和丙硫氧嘧啶（PROP）建立了定量苦味评价模型。该 BioET 的苦味特异性检测受到丙磺舒和 U-73122 的抑制，不受其他味觉模式或不激活 T2R38 的苦味配体的刺激。此外，通过结合不同的计算工具，我们设计了一种基于配体的虚拟筛选（LBVS）方案，以选择可能激活 T2R38 受体的配体。使用 LBVS 方案选择了三种计算预测的 T2R38 激动剂，BioET 对预测的激动剂有反应，验证了 LBVS 方案的能力。这项研究表明，这种独特的基于细胞的 BioET 为特异性检测可能用于药物研究和药物开发的含 N-C=S 化合物开辟了一条通用而有前途的途径。
