Healthcare Technology Institute, Institute of Translational Medicine, University of Birmingham, Birmingham B15 2TH, UK.
Centre for Pharmaceutical Engineering Science, School of Pharmacy and Medical Sciences, Faculty of Lifesciences, University of Bradford, Bradford BD7 1DP, UK.
Int J Mol Sci. 2023 Feb 8;24(4):3364. doi: 10.3390/ijms24043364.
This study reports results of a mechanical platform-based screening assay (MICA) to evaluate the remote activation of mechanosensitive ion channels. Here, we studied ERK pathway activation and the elevation in intracellular Ca levels in response to the MICA application using the Luciferase assay and Fluo-8AM assay, respectively. Functionalised magnetic nanoparticles (MNPs) targeting membrane-bound integrins and mechanosensitive TREK1 ion channels were studied with HEK293 cell lines under MICA application. The study demonstrated that active targeting of mechanosensitive integrins via RGD (Arginylglycylaspartic acid) motifs or TREK1 (KCNK2, potassium channel subfamily K member 2) ion channels can stimulate the ERK pathway and intracellular calcium levels compared to non-MICA controls. This screening assay offers a powerful tool, which aligns with existing high-throughput drug screening platforms for use in the assessment of drugs that interact with ion channels and influence ion channel-modulated diseases.
本研究报告了一种基于机械平台的筛选测定法(MICA)的研究结果,用于评估机械敏感离子通道的远程激活。在这里,我们分别使用荧光素酶测定法和 Fluo-8AM 测定法研究了 MICA 应用后 ERK 通路的激活和细胞内 Ca2+水平的升高。用针对膜结合整联蛋白和机械敏感 TREK1 离子通道的功能化磁性纳米颗粒(MNPs)在 MICA 应用下研究了 HEK293 细胞系。该研究表明,通过 RGD(精氨酸-甘氨酸-天冬氨酸)基序或 TREK1(KCNK2,钾通道亚家族 K 成员 2)离子通道对机械敏感整联蛋白的主动靶向,与非 MICA 对照相比,可以刺激 ERK 通路和细胞内钙水平。该筛选测定法提供了一种强大的工具,与现有的高通量药物筛选平台相匹配,可用于评估与离子通道相互作用并影响离子通道调节疾病的药物。