NPQ/Spexin the Endogenous Ligand for the Galanin Receptor 3

NPQ/Spexin 甘丙肽受体 3 的内源性配体

• 批准号: 9185000
• 负责人: Akihiko Ozawa
• 金额: $ 74.24万
• 依托单位: TORREY PINES INST FOR MOLECULAR STUDIES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9185000
• 关键词: Acute; Acute Pain; Adverse effects; Agonist; Alanine; Amino Acids; Analgesics; Applications Grants; Behavioral; Binding; Bioinformatics; Biological; Biological Assay; Brain; Brain region; Cardiovascular system; Cell physiology; Chemicals; Chinese Hamster Ovary Cell; Chronic; Chronic inflammatory pain; Computing Methodologies; Cyclic AMP; Development; Disease; Dopamine Agonists; Drug Industry; Drug Prescriptions; Electrophysiology (science); Family; Freund' s Adjuvant; G-Protein-Coupled Receptors; GALR3 Galanin Receptor; Galanin; Generations; Grant; In Vitro; Individual; Investigation; Kidney; Knockout Mice; Lead; Libraries; Ligands; Mediating; Melatonin; Messenger RNA; Methods; Molecular Target; Morphine; Mus; Natural Products; Neurons; Neuropeptide Receptor; Neuropeptides; Neurotransmitters; Nociception; Opiates; Opioid; Pain; Pain management; Peptide Receptor; Peptides; Perception; Peripheral; Pharmacotherapy; Physiological; Play; Preparation; Process; Property; Proprotein Convertase 1; Publishing; Receptor Activation; Recording of previous events; Reporting; Role; Scanning; Series; Slice; Smooth Muscle; Spinal; Spinal Cord; Spinal Ganglia; Spinal nerve structure; System; Testing; Validation; base; behavioral response; chronic neuropathic pain; chronic pain; combinatorial; disease mechanisms study; galanin receptor; gamma-Aminobutyric Acid; gastric fundus; immunoreactivity; in vitro Assay; in vivo; in vivo Model; inflammatory neuropathic pain; inflammatory pain; new therapeutic target; nociceptive response; novel; novel therapeutics; patch clamp; peptide analog; peptide hormone; pre-clinical; programs; prohormone; psychologic; receptor; receptor binding; research study; response; scaffold; screening; small molecule

Project Summary Neuropeptides are a class of neurotransmitters or peptide hormones that generally activate G-Protein Coupled Receptors (GPCRs) and are involved in a large number of physiological and psychological processes in the brain and in the periphery. One such neuropeptide that we have identified computationally we called NPQ and was called Spexin by another group that independently identified the same molecule by a different computational method. Peptides derived from preproNPQ/Spexin are found in very localized regions of the brain, and we have determined that they are involved in pain response as well as having renal and cardiovascular actions. Very recently, it was discovered that NPQ/spexin is in the galanin peptide family and binds to and activates the Galanin Receptor 3 (GALR3). In fact, as we have validated, NPQ activates GALR3 at a far lower concentration than galanin activates this receptor. Based upon this discovery, we have hypothesized that NPQ, rather than galanin, is the primary neuropeptide that mediates the actions of GALR3, and through GALR3, NPQ has a role in acute and chronic pain and is a potential new target for as non- addicting analgesic. To test these hypotheses, Specific Aim 1 will use a series of in vitro assays to examine the interaction between NPQ peptides and GALR3 in vitro. GALR3 containing CHO cells that we have produced will be used to examine the binding and functional activity of NPQ and NPQ 53-70. Aim 1 will also use patch clamp electrophysiology to examine the ability of NPQ to block GABA release in slices from the ventrolateral PAG, a region with high levels of NPQ and GALR3, and known to be involved in pain perception. Specific Aim 2 will use in vivo assays to directly examine the involvement of NPQ and GALR3 in pain using in vivo models of acute thermal and inflammatory pain, and chronic neuropathic and inflammatory pain. Specific Aim 3 will screen combinatorial libraries for hit/lead generationto identify new small molecule agonists andbegin the process of lead identification for the development of novel therapeutics. Specific Aim 4 will further validate this target by examining the immunohistochemical localization of peptides derived from proNPQ, as well as the prohormone (proNPQ) itself. This will be compared with the localization of GALR3 and galanin. To determine how NPQ and GALR3 are involved in chronic neuropathic and inflammatory pain, NPQ peptides and GALR3 immunoreactivity, as well as mRNA levels, will compared in naïve, spinal nerve ligated, and CFA treated mice. The involvement of GALR3 in the actions of NPQ will be verified by the use of GALR3 knockout mice. These experiments will verify the observation that NPQ activates GALR3, use in vitro and in vivo methods to examine how the NPQ/GALR3 system is involved in pain, and initiate a screening program to identify small molecule agonists as potential non-addicting analgesics through a novel GPCR target.

项目摘要 神经肽是一类神经递质或胡椒激素，通常激活G蛋白偶联 受体（GPCR），并参与了大量的生理和心理过程 大脑和周围。一种这样的神经肽，我们已经在计算上确定了NPQ，并且 另一组称为Spexin 计算方法。在prepronpq/spexin衍生的肽中发现在非常局部的区域 大脑，我们已经确定它们参与疼痛反应以及肾脏和 心血管动作。最近，发现NPQ/Spexin在Galanin Pepper家族中， 结合并激活Galanin受体3（GALR3）。实际上，正如我们已经验证的那样，NPQ激活GalR3 在浓度远低于galanin的情况下，激活该接收器。根据这一发现，我们有 假设NPQ而不是Galanin是介导GalR3的作用的主要神经肽， 通过GalR3，NPQ在急性和慢性疼痛中发挥作用，并且是非 - 加性镇痛。为了检验这些假设，具体目标1将使用一系列体外评估进行检查 NPQ肽与GalR3体外的相互作用。 GALR3包含我们具有的CHO单元 产生的将用于检查NPQ和NPQ 53-70的结合和功能活性。 AIM 1也将 使用斑块夹电生理学检查NPQ阻止GABA释放在切片中的能力 腹外侧PAG，一个具有高水平NPQ和GALR3的区域，已知参与疼痛感知。 特定目标2将使用体内测定直接检查NPQ和GalR3在疼痛中的参与 急性热和炎症性疼痛以及慢性神经性和炎症性疼痛的体内模型。具体的 AIM 3将筛选命中/铅生成的组合库，以识别新的小分子激动剂 并伸向新疗法发展的铅鉴定过程。具体目标4将 通过检查源自宠物的免疫组织化学定位，进一步验证了这一目标 PRONPQ以及激素（PRONPQ）本身。这将与galr3的本地化和 加拉宁。为了确定NPQ和GALR3如何参与慢性神经性和炎症性疼痛，NPQ 肽和GALR3免疫反应性以及mRNA水平将在幼稚的脊神经结扎中进行比较， 和CFA治疗的小鼠。 GalR3参与NPQ的作用将通过使用GalR3来验证 淘汰老鼠。这些实验将验证NPQ激活galr3，在体外和中使用的观察 体内方法检查NPQ/GALR3系统如何参与疼痛，并启动筛查程序 通过新型的GPCR靶标将小分子激动剂识别为潜在的非核心镇痛药。