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.

项目摘要瘙痒是一个复杂的生理过程，它结合了皮肤中感觉神经元刺激物的检测激活脊柱中间神经元，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）在每个瘙痒感和传播水平上都起着不可或缺的作用。虽然通常认为是脱敏和内在化的细胞表面受体配体结合，新证据已经确定了GPCR从内体信号的能力。但是，很少知道调节GPCR的内体信号的机制，并且对内体GPCR信号传导在瘙痒中的作用，或者内体GPCR是否是可行的治疗靶标痒。该提议假设：1。胃蛋白释放肽受体（GRPR）和神经激素1 受体（NK1R）是脊髓中瘙痒传播中的两个关键受体，可以募集并组装多种内体室的蛋白质复合物，促进内体信号传导并进行介导脊柱中间神经元的长时间过度刺激性。 GRPR和NK1R发出信号的能力内体导致阻断GRPR和NK1R的脊髓和内吞抑制剂的瘙痒传播内体信号传导可以抑制小鼠的刮擦行为。 2。靶向GRPR的内体信号传导与靶向细胞表面受体相比，使用纳米颗粒的NK1R是抑制瘙痒的更有效策略。 GRPR和NK1R的内体信号传导将在模型细胞系，脊柱中间神经元中进行表征介导瘙痒的传播和完整的动物。药物和遗传方法将用于抑制GRPR和NK1R的内体贩运。 AIM 1将表征内体的重要性 GRPR和NK1R在脊柱中间神经元中的运输和信号传播，用于传播瘙痒。的作用瘙痒中的内体信号传导将通过电生理学和瘙痒行为分析来解决动物。 AIM 2将表征GRPR访问内体的能力并组装多蛋白导致亚细胞特异性信号事件的复合物。这些信号传导复合物将使用具有高时空分辨率的晚期生物物理和成像方法。 AIM 3将使用高级化学生物学，纳米工程和纳米颗粒封装以提供GRPR和NK1R拮抗剂对于内体，探测GRPR和NK1R的内体信号在瘙痒传输中的重要性确定内体GPCR是否是可行的治疗靶标。