Lymphocyte Antigen 6 (Ly6) Proteins: New Players in Chronic Pain

淋巴细胞抗原 6 (Ly6) 蛋白：慢性疼痛的新参与者

• 批准号: 10784019
• 负责人: Kimberly Gomez
• 金额: $ 23.6万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-25 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10784019
• 关键词: Action Potentials; Affect; Afferent Neurons; Analgesics; Behavior; Behavioral; Binding; Biochemical; Biological Assay; Cell Communication; Cell Maturation; Cell Proliferation; Cells; Closure by clamp; Data; Dissociation; Down-Regulation; Electrophysiology (science); Evaluation; Extracellular Protein; Fingers; Fluorescence-Activated Cell Sorting; Gene Family; Generations; Genes; Genetic; Genus Lynx; Glycoproteins; Glycosylphosphatidylinositols; Goals; Human; Immune; In Situ Hybridization; Injury; Intrathecal Injections; Invaded; Knock-out; Knockout Mice; LY6E gene; Lead; Learning; Ligation; Link; Lipids; Lymphocyte antigen; Macrophage; Macrophage Activation; Malignant Neoplasms; Measures; Membrane; Modeling; Molecular; Mus; Nerve Fibers; Neurons; Neuropathy; Nicotinic Receptors; Nociception; Orthologous Gene; Pain; Pain Threshold; Pathologic; Patients; Pattern; Peptides; Physiological; Plasmids; Play; Population; Production; Protein Family; Proteins; Rattus; Regulation; Reporting; Rodent; Role; Shaker potassium channel; Signal Transduction; Small Interfering RNA; Snake Venoms; Sodium; Sodium Channel; Specificity; Spinal Cord; Spinal Ganglia; Stimulus; Sumoylation Pathway; Surface; Syndrome; Techniques; Testing; Time; Toxin; Transfection; Up-Regulation; Validation; afferent nerve; biophysical properties; cancer type; cell motility; chronic neuropathic pain; chronic pain; collapsin response mediator protein-2; confocal imaging; cytokine; design; extracellular; gain of function mutation; in vivo; mechanical allodynia; molecular sequence database; nerve injury; neuronal excitability; neurotransmission; neurotransmitter release; novel; overexpression; pain behavior; pain relief; pain signal; painful neuropathy; patch clamp; postsynaptic; protein protein interaction; response; sensory system; spared nerve; trafficking; transcriptome sequencing; ubiquitin-protein ligase; voltage

PROJECT SUMMARY Chronic pain is a pathological state where sensory neurons become hyperexcitable leading to nociceptive neurotransmission in the absence of a painful stimulus. Genetic and functional studies have established the voltage-gated sodium channel NaV1.7 as a major contributor to human pain signaling. Although the regulation of NaV1.7 is poorly understood, it is thought to involve mechanisms related to surface trafficking and regulation via protein-protein interactions. For instance, upregulation of SUMOylation of cytosolic collapsin response mediator protein 2 (CRMP2) and VGSC β-subunits, and downregulation of Nedd4-2 (a cytosolic E3 ubiquitin ligase) in a model of spared nerve injury induced chronic pain result in functional upregulation of NaV1.7 channels. My studies have identified lymphocyte antigen 6 (Ly6) proteins as a novel class of NaV1.7 channel modulators. Ly6 proteins are extracellular glycoproteins that are a hallmark of different types of cancer and have a role in cell proliferation, cell migration, cell–cell interactions, immune cell maturation, macrophage activation, and cytokine production. Ly6 proteins show structural resemblance to the three-fingered snake venom toxins that are known to modulate nicotinic acetylcholine receptors and voltage-gated sodium channels. Of these group of proteins, Ly6e and Lynx1 are common between humans and rodents. RNA-sequencing databases showed that both Ly6e and Lyn1 expression increases in different populations of dorsal root ganglia (DRG) neurons after nerve injury. Moreover, my preliminary findings show that: (i) there are higher Ly6e signal levels in DRGs after spared nerve injury (SNI); (ii) overexpression of Ly6e is associated with increased sodium currents in DRGs, and NaV1.7 currents in HEK cells; and (iii) intrathecal injection of Ly6e plasmid induces pain- like behaviors in naïve rats. These data led me to hypothesize that: (i) modulation of NaV1.7 channels by Ly6e and Lynx1 may lead to altered expression and activity of these channels during chronic pain, and that (ii) interfering with NaV1.7-Ly6e/Lynx1 interactions may relieve pain. Thus, the goals of this proposal are to investigate the role of Ly6 proteins (i) in sensory neurons and (ii) as molecular determinants of the altered functional activity of NaV1.7 channels in pain states. My Specific Aims are: (1) Investigate the physiological function(s) of Ly6e and Lynx1 in primary sensory neurons from rodents with and without nerve injury; (2) Identify and characterize Ly6e and Lynx1 as modulators of NaV1.7 channels in rodent and human dorsal root ganglia neurons; and (3) Identify specific interaction domain(s) in NaV1.7, Ly6e and Lynx1 and validation of in vivo target engagement. These studies are anticipated to advance our understanding of the role of Ly6e and Lynx1 in the sensory system, and their role as modulators of a key pain-associated voltage-gated sodium channel, NaV1.7.

项目摘要 慢性疼痛是一种病理状态，其中感觉神经元变得过度兴奋，导致伤害性疼痛。 在没有疼痛刺激的情况下神经传递。遗传和功能研究已经建立了 电压门控钠通道NaV1.7作为人类疼痛信号的主要贡献者。虽然规则 人们对NaV1.7了解甚少，认为它涉及与表面运输和调节有关的机制， 蛋白质相互作用例如，胞浆中的类胡萝卜素反应介质的SUMO化的上调， 蛋白2（CRMP 2）和VGSC β亚基的表达，以及Nedd 4 -2（一种细胞溶质E3泛素连接酶）在细胞凋亡中的下调。 备用神经损伤诱导的慢性疼痛模型导致NaV1.7通道的功能上调。我 研究已经鉴定出淋巴细胞抗原6（Ly 6）蛋白是一类新的NaV1.7通道调节剂。 Ly 6蛋白是细胞外糖蛋白，其是不同类型癌症的标志，并且在肿瘤的发生和发展中起作用。 细胞增殖、细胞迁移、细胞-细胞相互作用、免疫细胞成熟、巨噬细胞活化，以及 细胞因子产生。Ly 6蛋白与三指蛇毒毒素结构相似 已知其调节烟碱乙酰胆碱受体和电压门控钠通道。的 这组蛋白质Ly 6 e和Lynx 1在人类和啮齿类动物中是常见的。rna测序 数据库显示，Ly 6 e和Lyn 1的表达在不同的背根神经节群体中增加， (DRG)神经损伤后的神经元此外，我的初步研究结果表明：（一）有较高的Ly 6 e信号， 备用神经损伤（SNI）后DRG中的Ly 6 e水平;（ii）Ly 6 e的过表达与增加的钠水平相关。 DRG中的NaV1.7电流和HEK细胞中的NaV1.7电流;和（iii）鞘内注射Ly 6 e质粒诱导疼痛。 就像幼稚老鼠的行为一样。这些数据使我假设：（i）Ly 6 e对NaV1.7通道的调节 和Lynx 1可能导致慢性疼痛期间这些通道的表达和活性改变，以及（ii） 干扰NaV1.7-Ly 6 e/Lynx 1相互作用可减轻疼痛。因此，本提案的目标是 研究Ly 6蛋白（i）在感觉神经元中的作用和（ii）作为改变的神经元的分子决定因素。 NaV1.7通道在疼痛状态下的功能活性。我的具体目标是：（1）研究生理 Ly 6 e和Lynx 1在有和没有神经损伤的啮齿动物的初级感觉神经元中的功能;（2）鉴定 并将Ly 6 e和Lynx 1表征为啮齿动物和人类背根神经节中NaV1.7通道的调节剂 （3）鉴定NaV1.7、Ly 6 e和Lynx 1中的特异性相互作用结构域，并验证体内靶点 订婚这些研究预计将促进我们对Ly 6 e和Lynx 1在细胞凋亡中作用的理解。 感觉系统，以及它们作为关键疼痛相关电压门控钠通道NaV1.7的调节剂的作用。