Novel mechanism on subpopulation-dependent biased GPCR signaling in neurons

神经元亚群依赖性偏向 GPCR 信号传导的新机制

• 批准号: 10174956
• 负责人: YANG K XIANG
• 金额: $ 27.63万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10174956
• 关键词: Adrenergic Agents; Adrenergic Receptor; Adrenergic beta-Antagonists; Agonist; Alprenolol; Alzheimer' s Disease; Anxiety; Anxiety Disorders; Asthma; Attention; Attention deficit hyperactivity disorder; Biochemical; Biosensor; Cardiovascular Diseases; Cell membrane; Chronic Obstructive Airway Disease; Clinical; Crystallography; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Dendrites; Disease; Drug Receptors; Drug Targeting; Explosion; Family; Fluorescence Resonance Energy Transfer; Fractionation; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Glaucoma; Heart failure; Hippocampus (Brain); Hypertension; Image; Individual; Ion Channel; L-Type Calcium Channels; Learning; Ligands; Link; Mammalian Cell; Memory; Mental Depression; Metabolic syndrome; Modification; Nature; Neuraxis; Neurons; Norepinephrine; Peripheral; Phosphorylation; Phosphotransferases; Physiological; Play; Post-Traumatic Stress Disorders; Psyche structure; Resolution; Role; Signal Transduction; Sympathomimetics; Synapses; Synaptic plasticity; Testing; alertness; alpha-adrenergic receptor; base; beta-adrenergic receptor; carvedilol; cellular imaging; clinical application; clinically relevant; design; innovation; interest; neuronal excitability; novel; postsynaptic; receptor; response; side effect; tool; trafficking

Summary Adrenergic receptors (ARs) are a family of prototypical GPCRs linked to neuronal disorders, metabolic syndrome, and cardiovascular diseases. In the CNS, norepinephrine (NE) regulates attention and alertness. The β2AR is emerging as the prevalent postsynaptic NE effector at glutmatergic synapses, where it interacts with AMPAR, NMDAR and L-type Ca2+ channel Cav1.2 to modulate neuronal excitability, synaptic plasticity, and memory and learning. It is clinically relevant to understand NE-linked mental diseases such as depression, attention deficit hyperactivity disorder (ADHD), anxiety disorders (e.g., posttraumatic stress disorder, PTSD) and Alzheimer's disease. While β-blockers are used to treat a variety of peripheral diseases including heart failure, hypertension, glaucoma, asthma, and COPD, their clinical utility is hampered by the side effects including anxiety and depression. Recent explosion of crystallography study of ligand-GPCR interactions, there is still limited understanding on how a specific ligand leads to pleotropic cellular responses (including sides effects) of a GPCR. In this study, we hypothesize that a distinct subpopulation of PKA-phosphorylated β2ARs control LTCC activation in hippocampal neurons, which can be selectively activated by a set of biased ligands. We will test our hypothesis with following specific aims: Aim 1 is to test the hypothesis that β2AR can exist in distinct functional subpopulations in a single mammalian cell. We will use biochemical isolation/fractionation and super-resolution imaging to characterize distinct subcellular distribution of PKA-pβ2AR and GRK-pβ2AR. Aim 2 is to test the hypothesis that PKA-pβ2AR transduce biased signal through selectively modulation of ion channel activity at the plasma membrane (PM). Aim 3 is to test the hypothesis that sympathomimetic β-blockers act as biased ligands that selectively activate PKA-phosphorylated subpopulation of β2AR to activate ion channel at the PM. If successful, these aims will reveal a platform to understand the biased signaling induced by two distinct subpopulations of β2AR, and offering a new avenues for designing more efficacious β-AR drugs with fewer side effects in clinical applications.

概括 肾上腺素受体（ARS）是与神经元疾病有关的典型GPCR家族 综合征和心血管疾病。在中枢神经系统中，去甲肾上腺素（NE）调节注意力和机敏性。 β2AR正在作为谷物材料突触的普遍突触后NE效应子出现，在那里它相互作用 使用AMPAR，NMDAR和L型Ca2+通道CAV1.2调节神经元令人兴奋的突触可塑性， 以及记忆和学习。了解与NE连接的精神疾病（例如 抑郁，注意力缺陷多动症（ADHD），焦虑症（例如创伤后压力） 疾病，PTSD）和阿尔茨海默氏病。而β受体阻滞剂用于治疗各种外围 包括心力衰竭，高血压，青光眼，哮喘和COPD在内的疾病，其临床实用性为 受到副作用的阻碍，包括焦虑和抑郁。晶体学研究的最新爆炸 配体-GPCR相互作用，对特定配体如何导致多旋转的理解仍然有限 GPCR的细胞反应（包括侧面效应）。在这项研究中，我们假设一个独特的 海马神经元中PKA磷酸化的β2ARS控制LTCC激活的亚群，可以 通过一组有偏的配体选择性激活。我们将以以下特定目的检验我们的假设： 目的1是检验β2AR可以存在于单个功能亚群中的假设 哺乳动物细胞。我们将使用生化隔离/分馏和超分辨率成像 表征PKA-Pβ2AR和GRK-Pβ2AR的不同亚细胞分布。目标2是检验假设 PKA-Pβ2AR通过选择性调节离子通道活性在 质膜（PM）。 AIM 3是测试交感神经β受体阻滞剂的假设 选择性激活β2AR的PKA磷酸化亚群以激活离子通道的配体 下午。如果成功，这些目标将揭示一个平台，以了解两个由两个的偏见信号传导 β2AR的独特亚群，并提供新的途径，以设计更有效的β-AR药物 临床应用中的副作用较少。