Evaluating Cellular Impedance Assays for Detection of GPCR Pleiotropic Signaling and Functional Selectivity
Evaluating Cellular Impedance Assays for Detection of GPCR Pleiotropic Signaling and Functional Selectivity
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DOI：
10.1177/1087057108330115
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发表时间：
2009-03-01
影响因子：
--
通讯作者：
Scott, Clay W.
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文献类型：
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作者：
Peters, Matthew F.;Scott, Clay W.
G蛋白偶联受体可以偶联到不同细胞类型中的不同信号转导途径（称为细胞特异性信号传导），并且可以根据活化配体稳定的受体构象（功能选择性）激活不同的信号传导途径。这些概念为开发提高疗效和减少副作用的途径特异性药物提供了潜力。尽管有很大的兴趣，但功能选择性在药物发现中一直难以利用，部分原因是多个测定的负担。细胞阻抗测定使用一种新兴技术，可以在单一测定中定性区分Gs、GI/o和Gq信号传导，因此适合研究这些药理学概念。细胞阻抗证实了黑皮质素-4受体的细胞特异性Gs和Gq偶联以及大麻素-1（CB 1）受体的双重Gi和Gs信号传导。Gi与Gs信号传导的平衡取决于细胞系。在CB 1-HEKs中，GI和GS样响应相结合，产生一种新的阻抗曲线，证明了这些痕迹的动态性质。在U-2细胞和SK-N-MC细胞中观察到内源性D1受体的细胞特异性信号传导，但两种细胞系中部分和完全激动剂的药理学特征相似。我们的结论是，动态阻抗谱编码有价值的相对信号信息，是足够强大的，以帮助评估细胞特异性信号和功能的选择性。（Journal of Biomolecular Screening 2009：246-255）
G-protein-coupled receptors can couple to different signal transduction pathways in different cell types (termed cell-specific signaling) and can activate different signaling pathways depending on the receptor conformation(s) stabilized by the activating ligand (functional selectivity). These concepts offer potential for developing pathway-specific drugs that increase efficacy and reduce side effects. Despite significant interest, functional selectivity has been difficult to exploit in drug discovery, in part due to the burden of multiple assays. Cellular impedance assays use an emerging technology that can qualitatively distinguish Gs, Gi/o, and Gq signaling in a single assay and is thereby suited for studying these pharmacological concepts. Cellular impedance confirmed cell-specific Gs and Gq coupling for the melanocortin-4 receptor and dual Gi and Gs signaling with the cannabinoid-1 (CB1) receptor. The balance of Gi versus Gs signaling depended on the cell line. In CB1-HEKs, Gi-and Gs-like responses combined to yield a novel impedance profile demonstrating the dynamic nature of these traces. Cell-specific signaling was observed with endogenous D1 receptor in U-2 cells and SK-N-MC cells, yet the pharmacological profile of partial and full agonists was similar in both cell lines. We conclude that the dynamic impedance profile encodes valuable relative signaling information and is sufficiently robust to help evaluate cell-specific signaling and functional selectivity. (Journal of Biomolecular Screening 2009:246-255)