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.

项目总结/摘要 慢性瘙痒症是主要原因访问皮肤科医生和有毁灭性的影响， 生活质量尽管进行了广泛的研究，但慢性瘙痒的有效治疗仍然难以捉摸，部分原因是 因为我们对瘙痒背后的基本神经编码知之甚少。研究者的长期目标 研究计划是阐明神经机制的基础上处理瘙痒输入正常和 病理条件。有趣的是，慢性瘙痒症的一个标志是兴奋性神经递质的上调。 在脊髓内，有两种神经肽--P物质（SP）和胃泌素释放肽（GRP）--驱动瘙痒。 然而，神经肽调节脊髓回路产生瘙痒的机制尚不清楚。解决 这一知识的根本差距，这项建议将调查是否神经肽（SP和GRP）的作用， 与神经递质平行（在相同的目标上），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。具体来说，调查员 将测试这一假设，即瘙痒诱导神经肽SP和GRP通过不同的信号传导， 重新配置急性和持续性瘙痒的神经回路。在K99阶段，她将学习和应用切割- 边缘方法，从神经元超微结构到群体水平。目标1（K99）将比较脊柱 突触连接到神经肽连接使用多路电子显微镜。目标2（K99）将 确定由神经肽激活的脊髓网络与来自SP（和GRP）脊髓的突触传递 神经元使用双光子Ca 2+成像与离体体感准备相结合。的一部分 K99阶段还将致力于学习实现机器学习算法，以检测和量化 老鼠痒行为，以及职业发展活动，如获得领导和指导技能， 在会议上发表演讲，以扩大申请人的专业网络。目标3（R 00）将决定是否 SP和GRP信号传导是异常自发性脊髓活动的表现所必需的（目的 3A）和与使用药理学抑制的持续性瘙痒相关的瘙痒行为升高（目的3B） 战略布局这些研究将利用她在双光子Ca 2+成像和自动行为成像方面的新专业知识。 在K99阶段开发的分析管道结合她在啮齿动物瘙痒方面的强大背景 模型总之，这些实验将为脊髓瘙痒中的神经肽信号提供基本的见解 传递和确定脊髓神经肽信号传导在持续性瘙痒中的必要性。的成功 职业奖励培训计划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 Research.完成本计划将准备申请人领导一个创新的研究计划， 独立调查员