Mechanisms of Endosomal Signaling of Itch

瘙痒的内体信号传导机制

基本信息

批准号：
10541169
负责人：
Dane D Jensen
金额：
$ 40.13万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-15 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10541169
关键词：
Address Afferent Neurons Agonist Animals Anxiety Behavior Behavioral Assay Biology Bioluminescence Bombesin Receptor Cell Line Cell Nucleus Cell Surface Receptors Cell membrane Cell model Cells Chemicals Clathrin Complex Confocal Microscopy Cytosol Detection Dynamin Electrophysiology (science)Encapsulated Endocytosis Endosomes Energy Transfer Event Excision FDA approved Family Fluorescence Resonance Energy Transfer G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors Gastrin releasing peptide Interneurons Irritants Knowledge Ligand Binding Link Mediating Mental Depression Modality Modeling Multiprotein Complexes Mus Neural Pathways Neurons Pathologic Pathologic Processes Pathway interactions Peptide Receptor Pharmaceutical Preparations Pharmacologic Substance Physiological Physiological Processes Play Pruritus Publishing Quality of life Receptor Activation Receptor Signaling Reporting Research Project Grants Resolution Role Sensory Signal Transduction Skin Slice Small Interfering RNA Spinal Spinal Cord Stimulus Stress Substance P Suicide TACR1 gene Testing Therapeutic Vertebral column antagonist biophysical techniques chronic itch chronic pain confocal imaging conventional therapy desensitization design experience genetic approach imaging approach improved inhibitor insight knock-down method development mouse model nanoengineering nanoparticle nanoparticle delivery novel therapeutics peptide P pharmacologic prevent receptor recruit response sensory stimulus signal recognition particle receptor spatiotemporal targeted treatment therapeutic target therapy design trafficking transmission process

项目摘要

Project Summary Itch is a complex physiological process that incorporates detection of irritants by sensory neurons in the skin which activate spinal interneurons and ultimately, cortical projection neurons to generate a response. G protein-coupled receptors (GPCRs) play an integral role at each level of itch sensation and transmission. Although conventionally considered cell surface receptors that are desensitized and internalized following ligand binding, new evidence has established the ability of GPCRs to signal from endosomes. However, little is known about the mechanisms that regulate endosomal signaling of GPCRs and nothing is known about the role of endosomal GPCRs signaling in itch or whether endosomal GPCRs are a viable therapeutic target for itch. This proposal hypothesizes that: 1. Gastrin releasing peptide receptor (GRPR) and neurokinin 1 receptor (NK1R), two key receptors in itch transmission in the spinal cord, can recruit and assemble multi- protein complexes from the endosomal compartment that facilitate endosomal signaling and mediate prolonged hyperexcitability of spinal interneurons. That the ability of GRPR and NK1R to signaling from endosomes leads to itch transmission in the spinal cord and endocytic inhibitors that block GRPR and NK1R endosomal signaling can inhibit scratching behavior in mice. 2. Targeting endosomal signaling of GRPR and NK1R using nanoparticles is a more effective strategy in inhibiting itch than targeting cell surface receptors. Endosomal signaling of GRPR and NK1R will be characterized in model cell lines, spinal interneurons that mediate itch transmission, and in intact animals. Pharmaceutical and genetic approaches will be used to inhibit endosomal trafficking of GRPR and NK1R. Aim 1 will characterize the importance of endosomal trafficking and signaling of GRPR and NK1R in spinal interneurons for the transmission of itch. The role of endosomal signaling in itch will be addressed by electrophysiology, and by itch behavioral assays in intact animals. Aim 2 will characterize the ability of GRPR to traffic to endosomes and assemble the multi-protein complexes that result in subcellular specific signaling events. These signaling complexes will be studied using advanced biophysical and imaging approaches with high spatiotemporal resolution. Aim 3 will use advanced chemical biology, nanoengineering and nanoparticle encapsulation to deliver GRPR and NK1R antagonists to endosomes, probing the importance of endosomal signaling of GRPR and NK1R in itch transmission and to determine whether endosomal GPCRs are a viable therapeutic target.

项目概要瘙痒是一个复杂的生理过程，其中包括皮肤感觉神经元对刺激物的检测它激活脊髓中间神经元，并最终激活皮质投射神经元以产生反应。 G 蛋白质偶联受体（GPCR）在瘙痒感和传播的各个层面都发挥着不可或缺的作用。尽管传统上认为细胞表面受体在以下情况下脱敏和内化：配体结合，新证据证实了 GPCR 从内体发出信号的能力。然而，很少已知调节 GPCR 内体信号传导的机制，但对此一无所知内体 GPCR 信号传导在瘙痒中的作用或内体 GPCR 是否是可行的治疗靶点为了痒。该提案假设： 1. 胃泌素释放肽受体 (GRPR) 和神经激肽 1 受体（NK1R）是脊髓中瘙痒传播的两个关键受体，可以招募和组装多种来自内体区室的蛋白质复合物，促进内体信号传导并介导脊髓中间神经元长期过度兴奋。 GRPR 和 NK1R 传递信号的能力内体导致脊髓中的瘙痒传播以及阻断 GRPR 和 NK1R 的内吞抑制剂内体信号传导可以抑制小鼠的抓挠行为。 2. 靶向 GRPR 的内体信号传导和使用纳米颗粒的 NK1R 是一种比靶向细胞表面受体更有效的抑制瘙痒策略。 GRPR 和 NK1R 的内体信号传导将在模型细胞系、脊髓中间神经元中进行表征介导瘙痒传播，并且在完整的动物中。药物和遗传学方法将用于抑制 GRPR 和 NK1R 的内体运输。目标 1 将描述内体的重要性 GRPR 和 NK1R 在脊髓中间神经元中的运输和信号传导，用于传递瘙痒。的作用瘙痒中的内体信号传导将通过电生理学和完整的瘙痒行为测定来解决动物。目标 2 将表征 GRPR 运输至内体并组装多蛋白的能力导致亚细胞特异性信号传导事件的复合物。这些信号复合物将使用以下方法进行研究具有高时空分辨率的先进生物物理和成像方法。目标3将使用先进的化学生物学、纳米工程和纳米颗粒封装以提供 GRPR 和 NK1R 拮抗剂内体，探讨 GRPR 和 NK1R 内体信号传导在瘙痒传播中的重要性确定内体 GPCR 是否是可行的治疗靶点。