Targeted Genetic Silencing of Sensory Neurons to Inhibit Pain and Itch

感觉神经元的靶向基因沉默以抑制疼痛和瘙痒

• 批准号: 8378003
• 负责人: QIUFU MA
• 金额: $ 26.53万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8378003
• 关键词: 4 hydroxynonenal; Ablation; Acute; Acute Pain; Address; Adopted; Afferent Neurons; Afferent Pathways; Agonist; Analgesics; Animals; Atopic Dermatitis; Axon; Behavioral; Body Temperature; Calcium Channel Blockers; Capsaicin; Cells; Charge; Chemicals; Chronic; Data; Genetic; Glutamates; Heating; Inflammation; Inflammatory; Ion Channel; Label; Lidocaine; Lipids; Local anesthesia; Mechanics; Mediating; Modeling; Mus; Nerve; Neurons; Nociception; Nociceptors; Pain; Pain management; Pathway interactions; Peripheral; Principal Investigator; Protons; Rattus; Role; Side; Sodium; Sodium Channel Blockers; Stimulus; Synaptic Transmission; System; TRP channel; TRPV1 gene; Testing; The Sun; Therapeutic; Variant; Vesicle; Work; bean; behavior measurement; chronic pain; design; in vivo; inflammatory neuropathic pain; ion channel blocker; mustard oil; nerve injury; neuromechanism; novel strategies; novel therapeutic intervention; novel therapeutics; pain behavior; pain inhibition; painful neuropathy; patch clamp; programs; response; targeted delivery; transmission process

The specific contribufion of different primary sensory neurons in producing pain and itch will be invesfigated by targeted delivery of an impermeant cafionic sodium channel blocker (QX-314, a quaternary derivative of lidocaine) through large-pore ion channels differenfially expressed on afferents. The aim is to electrically silence subsets of axons using combinafions of QX-314 and large-pore ion channel agonists. We have validated this strategy for TRPV1-expressing nociceptors using capsaicin and QX-314 to produce a long lasfing "pain-specific" local anesthesia, and our pilot data suggest that TRPAl and P2X3 activafion by mustard oil and ATP also allow QX-314 permeation into DRG neurons. We will confirm this using whole cell patch clamp recordings in DRG neurons. The strategy will then be used in vivo to determine the funcfional effects on nociception of blocking TRPVl, TRPAl and P2X3 expressing nociceptors in naive rats and mice, measuring behavioral deficits in response to speciflc mechanical, thermal and chemical sfimuli. The afferent silencing strategy will also be used to idenfify which large-pore ion channel expressing primary afferents contribute to histaminergic and non-histaminergic itch. The presence of silenced afferents after administering QX-314 alone will be used to establish if large-pore ion channels are consfitufively acfivated by endogenous agonists during peripheral inflammafion, after nerve injury, in acute and chronic itch, when, and where. Our pilot data show that QX-314 blocks nociceptor terminals in the presence of inflammafion but not in naive animals. We will now explore if QX314 administered to peripheral terminals or nerves alters different modalifies of pain sensitivity in soft fissue, incisional, arthrific, and peripheral neuritic inflammatory models, as well as in neuropathic pain and pruritic atopic dermafitis models. These strategies will help both identify the cellular mechanisms of nocicepfive, inflammatory and neuropathic pain, acute and chronic itch and develop opportunifies for targeted novel therapeutic interventions to treat these condifions. RELEV/VNCE (See instmctions): We propose to develop a new analgesic strategy; targeting the pain pathway in the periphery using large- pore lon channels specific to the system as a means of delivering a sodium channel blocker only into defined sets of neurons. The proposal will test the potential of this appraoch for acute and chronic pain therapy.

将研究不同初级感觉神经元在产生疼痛和瘙痒中的具体贡献 通过靶向递送不渗透性钙离子钠通道阻滞剂（QX-314，一种四元衍生物） 利多卡因）通过大孔离子通道在传入神经上差异表达。其目的是通过电 使用 QX-314 和大孔离子通道激动剂的组合沉默轴突子集。我们有 使用辣椒素和 QX-314 验证了表达 TRPV1 的伤害感受器的这一策略，以产生长 激光“疼痛特异性”局部麻醉，我们的试验数据表明 TRPA1 和 P2X3 激活 芥末油和 ATP 还允许 QX-314 渗透到 DRG 神经元中。我们将使用全细胞来确认这一点 DRG 神经元中的膜片钳记录。然后该策略将用于体内以确定功能 阻断TRPV1、TRPA1和P2X3表达伤害感受器在幼稚大鼠和小鼠中对伤害感受的影响， 测量对特定机械、热和化学刺激的行为缺陷。传入的 沉默策略还将用于识别表达初级传入的大孔离子通道 导致组胺能和非组胺能瘙痒。给药后存在传入沉默 单独使用 QX-314 来确定大孔离子通道是否被内源性激活 外周炎症期间、神经损伤后、急性和慢性瘙痒时、何时何地使用激动剂。我们的 试验数据表明，QX-314 在存在炎症的情况下会阻断伤害感受器末梢，但在幼稚的情况下则不会 动物。我们现在将探讨将 QX314 施用到外周末梢或神经是否会改变不同的 改善软组织、切口、关节炎和周围神经炎炎症模型中的疼痛敏感性， 以及神经性疼痛和瘙痒性特应性皮炎模型。这些策略将有助于双方确定 伤害性疼痛、炎症性疼痛和神经性疼痛、急性和慢性瘙痒的细胞机制 开发有针对性的新型治疗干预措施来治疗这些病症的机会。 RELEV/VNCE（参见说明）： 我们建议制定新的镇痛策略；使用大的靶向周围疼痛通路 系统特有的孔离子通道作为仅将钠通道阻滞剂递送到指定的区域的手段 神经元组。该提案将测试这种方法用于急性和慢性疼痛治疗的潜力。