Mechanisms of Endosomal Signaling of Itch

瘙痒的内体信号传导机制

• 批准号: 10344775
• 负责人: Dane D Jensen
• 金额: $ 39.15万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 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/10344775
• 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; Depression and Suicide; 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; Modality; Modeling; Multiprotein Complexes; Mus; Neural Pathways; Neurons; Pathologic; Pathologic Processes; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; 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; Substance P Receptor; Testing; Therapeutic; 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; prevent; receptor; recruit; response; sensory stimulus; signal recognition particle receptor; spatiotemporal; targeted treatment; therapeutic target; therapy design; trafficking; transmission process

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，包括皮肤感觉神经元对刺激物的检测 其激活脊髓中间神经元并最终激活皮质投射神经元以产生响应。G 蛋白偶联受体（GPCR）在瘙痒感觉和传递的各个水平上起着不可或缺的作用。 尽管传统上认为细胞表面受体是脱敏和内化后， 配体结合，新的证据已经确立了GPCR从内体发出信号的能力。不过小 已知调节GPCR内体信号传导的机制， 内体GPCR信号传导在瘙痒中作用或内体GPCR是否是可行的治疗靶点 治痒。该提案假设：1。胃泌素释放肽受体与神经激肽1 受体（NK1R）是脊髓中瘙痒传递的两个关键受体，可以募集和组装多个 来自内体区室的蛋白质复合物，其促进内体信号传导并介导 延长脊髓中间神经元的过度兴奋。GRPR和NK1R的信号传导能力， 内体导致脊髓中的瘙痒传递和阻断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， 使用纳米颗粒的NK1R是比靶向细胞表面受体更有效的抑制瘙痒的策略。 GRPR和NK1R的内体信号传导将在模型细胞系、 介导瘙痒传播，并在完整的动物。药物和遗传方法将用于 抑制GRPR和NK1R内体运输。目的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瘙痒的传递。的作用 瘙痒中的内体信号传导将通过电生理学和完整的小鼠中的瘙痒行为测定来解决。 动物目的2将描述GRPR运输到内体并组装多蛋白的能力 导致亚细胞特异性信号事件的复合物。这些信号复合物将使用 先进的生物物理和成像方法，具有高时空分辨率。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，以递送GRPR和NK1R拮抗剂 内体，探测GRPR和NK1R的内体信号传导在瘙痒传递中的重要性， 以确定内体GPCR是否是可行的治疗靶点。