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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要慢性疼痛是一种病理状态，感觉神经元变得过度兴奋，导致伤害性感受遗传和功能研究已经确定了在没有疼痛刺激的情况下的神经传递。电压门控钠通道 NaV1.7 作为人类疼痛信号传导的主要贡献者。 NaV1.7 人们知之甚少，它被认为涉及与表面贩运和监管相关的机制例如，胞质塌陷素反应介质的 SUMO 化上调。蛋白 2 (CRMP2) 和 VGSC β 亚基，以及 Nedd4-2（一种胞质 E3 泛素连接酶）的下调神经损伤引起的慢性疼痛模型导致 NaV1.7 通道功能上调。研究已确定淋巴细胞抗原 6 (Ly6) 蛋白是一类新型 NaV1.7 通道调节剂。 Ly6 蛋白是细胞外糖蛋白，是不同类型癌症的标志，并在细胞增殖、细胞迁移、细胞间相互作用、免疫细胞成熟、巨噬细胞激活和 Ly6 蛋白与三指蛇毒毒素的结构相似。已知可调节烟碱乙酰胆碱受体和电压门控钠通道。这组蛋白质 Ly6e 和 Lynx1 在人类和啮齿动物中很常见。数据库显示 Ly6e 和 Lyn1 表达在不同背根神经节群体中增加此外，我的初步研究结果表明：（i）存在更高的 Ly6e 信号。幸存神经损伤 (SNI) 后 DRG 中的水平；(ii) Ly6e 的过度表达与钠含量增加相关 DRG 中的电流和 HEK 细胞中的 NaV1.7 电流；以及 (iii) 鞘内注射 Ly6e 质粒诱导疼痛这些数据让我发现：(i) Ly6e 对 NaV1.7 通道的调节。 Lynx1 可能会在慢性疼痛期间导致这些通道的表达和活动，并且 (ii) 干扰 NaV1.7-Ly6e/Lynx1 相互作用可能会减轻疼痛。因此，该提案的目标是研究 Ly6 蛋白 (i) 在感觉神经元中的作用以及 (ii) 作为感觉神经元的分子决定因素 NaV1.7 通道在疼痛状态下的功能活动我的具体目标是：（1）研究生理学。 Ly6e 和 Lynx1 在有和没有神经损伤的啮齿动物初级感觉神经元中的功能 (2) 识别并将 Ly6e 和 Lynx1 描述为啮齿动物和人类背根神经节 NaV1.7 通道的调节剂 (3) 识别 NaV1.7、Ly6e 和 Lynx1 中的特定相互作用域并验证体内靶点这些研究预计将增进我们对 Ly6e 和 Lynx1 在参与中的作用的理解。感觉系统，及其作为关键疼痛相关电压门控钠通道 NaV1.7 调节剂的作用。