Understanding spinal neuropeptide signaling in itch

了解瘙痒中的脊髓神经肽信号传导

• 批准号: 10426855
• 负责人: Tayler Sheahan
• 金额: $ 12.22万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10426855
• 关键词: Acute; Address; Algorithmic Analysis; Anatomy; Award; Behavior; Behavioral; Bombesin Receptor; Code; Collaborations; Dermatologist; Development; Disease; Dorsal; Electron Microscopy; Excitatory Synapse; Gastrin releasing peptide; Glutamates; Goals; Gold; Health; Image; Knowledge; Label; Lead; Leadership; Learning; Left; Mentorship; Modeling; Mus; Nervous system structure; Neurons; Neuropeptides; Neurotransmitters; Pain Clinics; Pain Research; Pathologic; Patients; Peptide Signal Sequences; Pharmacology; Phase; Population; Preparation; Pruritus; Quality of life; Research; Research Personnel; Rodent; Role; Signal Transduction; Skin; Spinal; Spinal Cord; Spinal nerve structure; Stimulus; Substance P; Substance P Receptor; Sum; Synapses; Synaptic Transmission; TACR1 gene; Testing; Time; Training; Up-Regulation; Visit; analysis pipeline; behavioral pharmacology; career; career development; career networking; chronic itch; effective therapy; experimental study; innovation; insight; machine learning algorithm; mouse model; neural circuit; neuromechanism; neurotransmission; novel therapeutics; programs; receptor; relating to nervous system; skills; somatosensory; success; symposium; training opportunity; transmission process; two-photon

PROJECT SUMMARY/ABSTRACT Chronic itch disorders are the primary reason for visits to dermatologists and have devasting effects on patient quality of life. Despite extensive research efforts, an effective treatment for chronic itch remains elusive, in part because the basic neural coding that underlies itch is poorly understood. The long-term goal of the investigator’s research program is to elucidate the neural mechanisms that underlie processing of itch input under normal and pathological conditions. Interestingly, one hallmark of chronic itch disorders is the upregulation of excitatory neuropeptides that drive itch – substance P (SP) and gastrin-releasing peptide (GRP) – within the spinal cord. However, the mechanism by which neuropeptides modulate spinal circuits to produce itch is unclear. To address this fundamental gap in knowledge, this proposal will investigate whether neuropeptides (SP and GRP) act in parallel with neurotransmitters (on the same targets) or if neuropeptide signaling diverges from neurotransmission (engage distinct targets), thereby reconfiguring spinal circuits. Specifically, the investigator will test the hypothesis that the itch-inducing neuropeptides SP and GRP act via divergent signaling to reconfigure neural circuits in acute and persistent itch. During the K99 Phase, she will learn and apply cutting- edge approaches that span from the neuron ultrastructural to population level. Aim 1 (K99) will compare spinal synaptic connectivity to neuropeptide connectivity using multiplexed electron microscopy. Aim 2 (K99) will identify the spinal networks activated by neuropeptide versus synaptic transmission from SP (and GRP) spinal neurons using two-photon Ca2+ imaging in combination with an ex vivo somatosensory preparation. A portion of the K99 phase will also be dedicated to learning to implement machine learning algorithms to detect and quantify mouse itch behaviors, and to career development activities such as gaining leadership and mentorship skills and presenting at conferences to expand the applicant’s professional network. Aim 3 (R00) will determine whether the SP and GRP signaling are required for the manifestation of aberrant spontaneous spinal cord activity (Aim 3A) and elevated itch behaviors (Aim 3B) associated with persistent itch using pharmacological inhibition strategies. These studies will leverage her new expertise in two-photon Ca2+ imaging and automated behavioral analysis pipelines developed during the K99 phase in combination with her strong background in rodent itch models. Together, these experiments will provide fundamental insights into neuropeptide signaling in spinal itch transmission and identify the necessity of spinal neuropeptide signaling in persistent itch. The success of this career award training plan, which includes activities for scientific and career development, will be aided by the scientific expertise, collaboration, and educational opportunities offered by the Pittsburgh Center for Pain Research. Completing this plan will prepare the applicant to lead an innovative research program as an independent investigator.

项目概要/摘要 慢性瘙痒症是去看皮肤科医生的主要原因，并对患者造成毁灭性影响 尽管进行了大量的研究工作，但慢性瘙痒的有效治疗方法仍然难以实现。 因为人们对瘙痒的基本神经编码知之甚少。研究者的长期目标。 研究计划旨在阐明正常和不同情况下处理瘙痒输入的神经机制。 慢性瘙痒症的同义病理特征之一是兴奋性上调。 脊髓内引起瘙痒的神经肽——P 物质 (SP) 和胃泌素释放肽 (GRP)。 然而，神经肽调节脊髓回路产生瘙痒的机制尚不清楚。 鉴于这一知识上的根本差距，该提案将研究神经肽（SP 和 GRP）是否在 与神经递质平行（在同一目标上），或者如果神经肽信号传导与神经递质不同 神经传递（参与不同的目标），从而重新配置脊髓回路，特别是研究者。 将检验以下假设：诱发瘙痒的神经肽 SP 和 GRP 通过不同的信号传导发挥作用 重新配置急性和持续性瘙痒的神经回路 在 K99 阶段，她将学习并应用切割。 从神经元超微结构到群体水平的边缘方法将比较脊柱。 使用多重电子显微镜观察突触与神经肽的连接。 识别神经肽激活的脊髓网络与 SP（和 GRP）脊髓的突触传递 使用双光子 Ca2+ 成像结合离体体感制剂的神经元。 K99阶段还将致力于学习实施机器学习算法来检测和量化 鼠标痒行为，以及职业发展活动，例如获得领导和指导技能以及 目标 3 (R00) 将决定是否在会议上进行演讲以扩大申请人的专业网络。 SP 和 GRP 信号传导是异常自发脊髓活动表现所必需的（目的 3A）和与使用药物抑制持续瘙痒相关的瘙痒行为升高（目标 3B） 这些研究将利用她在双光子 Ca2+ 成像和自动化行为方面的新专业知识。 在 K99 阶段结合她在啮齿动物瘙痒方面的深厚背景开发的分析流程 总之，这些实验将为脊髓瘙痒中的神经肽信号传导提供基础见解。 传输并确定脊髓神经肽信号传导在持续性瘙痒中的必要性。 职业奖励培训计划，其中包括科学和职业发展活动，将得到 匹兹堡疼痛中心提供的科学专业知识、合作和教育机会 完成该计划将使申请人做好领导创新研究计划的准备。 独立调查员。