Measurement of GPCR-mediated thallium flux:GIRK (RMI)

GPCR 介导的铊通量的测量：GIRK (RMI)

• 批准号: 7057949
• 负责人: COLLEEN M NISWENDER
• 金额: $ 0.46万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2006-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7057949
• 关键词: adrenergic agents; alpha adrenergic receptor; antiadrenergic agents; biotechnology; cell surface receptors; drug screening /evaluation; high throughput technology; potassium channel; receptor sensitivity; technology /technique development; thallium

DESCRIPTION (provided by applicant): Pharmacological agents targeting G-protein coupled receptors and ion channels represent promising clinical agents spanning a multitude of human diseases. We have developed a screening strategy for Gi/o-linked receptors by measuring thallium flux through G protein regulated Inwardly Rectifying Potassium (K+) (GIRK) channels. Using this technique in HEK cells stably expressing GIRK 1 and 2 channels, we have generated preliminary data using muscarinic and adrenergic agonists and antagonists and have observed dose dependent regulation of these channels by endogenous M2/M4 receptors and ?2C adrenergic receptor selective agents. We have adapted this assay to a 384 well plate format and have performed initial Z' experiments that indicate that the assay is amenable to HTS. We propose to conduct a high throughput screen for novel agonists at the ?2C receptor; in the course of these studies, it is anticipated that we will identify novel agonists of GIRK 1/2 channels as well. Very few specific pharmacological tools exist targeting the ?2C receptor and due to its proposed role in psychiatric disorders and normal cardiac function it is anticipated that agonists targeting this receptor would provide critical tools to test ?2C receptor function in vivo. GIRK1/2 channels are known to be involved in neuronal excitability and it has been proposed that agonists for this receptor may provide new therapeutic avenues for epilepsy and pain. Lay summary: G-protein coupled receptors (GPCRs) and ion channels represent accessible therapeutic targets in human disease. We have developed a new assay, amenable to high throughput strategies, which can be used to screen large chemical libraries for compounds acting within the signal transduction pathway from receptor to ion channel. We propose to use this assay to identify compounds interacting with the ?2C adrenergic receptor as well as agents directly modulating the potassium channel employed in our assay system. Identified compounds will provide new pharmacological tools to test the role of the ?2C adrenergic receptor in psychiatric and cardiovascular disorders and the role of GIRK 1/2 channels in the modulation of neuronal excitability.

描述（由申请人提供）：靶向G蛋白偶联受体和离子通道的药物代表了跨越多种人类疾病的有前途的临床药物。我们通过测量通过 G 蛋白调节的内向整流钾 (K+) (GIRK) 通道的铊通量，制定了一种 Gi/o 连接受体的筛选策略。在稳定表达 GIRK 1 和 2 通道的 HEK 细胞中使用该技术，我们使用毒蕈碱和肾上腺素激动剂和拮抗剂生成了初步数据，并观察到内源性 M2/M4 受体和 α2C 肾上腺素受体选择性剂对这些通道的剂量依赖性调节。我们已将此测定法调整为 384 孔板格式，并进行了初始 Z' 实验，表明该测定法适用于 HTS。我们建议对 2C 受体的新型激动剂进行高通量筛选；在这些研究过程中，预计我们还将鉴定 GIRK 1/2 通道的新型激动剂。针对β2C受体的特异性药理学工具很少，并且由于其在精神疾病和正常心脏功能中的作用，预计针对该受体的激动剂将提供体内测试β2C受体功能的关键工具。已知 GIRK1/2 通道与神经元兴奋性有关，并且有人提出该受体的激动剂可能为癫痫和疼痛提供新的治疗途径。简单总结：G 蛋白偶联受体 (GPCR) 和离子通道代表了人类疾病的可行治疗靶点。我们开发了一种新的检测方法，适用于高通量策略，可用于筛选大型化学库中在从受体到离子通道的信号转导途径中起作用的化合物。我们建议使用该测定法来鉴定与 2C 肾上腺素能受体相互作用的化合物以及直接调节我们的测定系统中使用的钾通道的药物。鉴定出的化合物将提供新的药理学工具来测试 2C 肾上腺素受体在精神和心血管疾病中的作用以及 GIRK 1/2 通道在神经元兴奋性调节中的作用。