Bias Analyses of Preclinical and Clinical D2 Dopamine Ligands: Studies with Immediate and Complex Signaling Pathways
Bias Analyses of Preclinical and Clinical D2 Dopamine Ligands: Studies with Immediate and Complex Signaling Pathways
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DOI：
10.1124/jpet.114.220293
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发表时间：
2015-03-01
影响因子：
3.5
通讯作者：
Watts, Val J.
中科院分区：
文献类型：
--
作者：
Brust, Tarsis F.;Hayes, Michael P.;Watts, Val J.
G 蛋白偶联受体 (GPCR) 通常激活多个信号传导途径，配体可能通过单个途径引发功能反应。这些独特的反应为偏向或功能选择性配体提供了机会，可以优先调节一种信号传导途径而不是另一种信号传导途径。对多个 GPCR 的研究表明，选择性激活 GPCR 下游信号通路可能会带来更安全、更有效的药物治疗。多巴胺 D-2 受体 (D2R) 是帕金森病和精神分裂症治疗的主要药物靶点之一。最近的研究表明，选择性调节 D2R 下游的单个信号通路可能会产生更安全的抗精神病药物。在本研究中，检查了 D2R 的直接效应器（即 G α(i/o)、G β γ、β-抑制蛋白募集）和更复杂的信号通路（即细胞外信号调节激酶磷酸化、异源致敏和动态质量再分配）对一系列 D2R 配体的反应。这是使用 PathHunter beta-Arrestin GPCR 检测平台中稳定表达人 D-2L 多巴胺受体的中国仓鼠卵巢细胞完成的。使用统一的细胞背景旨在消除与细胞间变异相关的潜在混杂因素，包括受体的表达水平以及信号转导的其他成分，包括 G 蛋白亚基。在该细胞模型中的每个信号通路中评估了几种特征明确且临床相关的 D2R 配体。比较了最常用的测量配体偏差的方法。功能选择性分析也被用作探索直接 D2R 效应器对激活更复杂信号通路的相对贡献的工具。
G protein-coupled receptors (GPCRs) often activate multiple signaling pathways, and ligands may evoke functional responses through individual pathways. These unique responses provide opportunities for biased or functionally selective ligands to preferentially modulate one signaling pathway over another. Studies with several GPCRs have suggested that selective activation of signaling pathways downstream of a GPCR may lead to safer and more effective drug therapies. The dopamine D-2 receptor (D2R) is one of the main drug targets in the therapies for Parkinson's disease and schizophrenia. Recent studies suggest that selective modulation of individual signaling pathways downstream of the D2R may lead to safer antipsychotic drugs. In the present study, immediate effectors of the D2R (i.e., G alpha(i/o), G beta gamma, beta-arrestin recruitment) and more complex signaling pathways (i.e., extracellular signal-regulated kinase phosphorylation, heterologous sensitization, and dynamic mass redistribution) were examined in response to a series of D2R ligands. This was accomplished using Chinese hamster ovary cells stably expressing the human D-2L dopamine receptor in the PathHunter beta-Arrestin GPCR Assay Platform. The use of a uniform cellular background was designed to eliminate potential confounds associated with cell-to-cell variability, including expression levels of receptor as well as other components of signal transduction, including G protein subunits. Several well characterized and clinically relevant D2R ligands were evaluated across each signaling pathway in this cellular model. The most commonly used methods to measure ligand bias were compared. Functional selectivity analyses were also used as tools to explore the relative contribution of immediate D2R effectors for the activation of more complex signaling pathways.