Project 1: Cao

项目一：曹

• 批准号: 8883622
• 负责人: Ling Cao
• 金额: $ 20.17万
• 依托单位: UNIVERSITY OF NEW ENGLAND
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8883622
• 关键词: Affect; Afferent Neurons; Behavioral; Biological Assay; CCL17 gene; CCL2 gene; CCL22 gene; CCL3 gene; CXCL1 gene; CXCL10 gene; Calcitonin Gene-Related Peptide; Calcitonin-Gene Related Peptide Receptor; Caliber; Capsaicin; Coculture Techniques; Data; Development; Dose; Enzyme-Linked Immunosorbent Assay; Event; Functional disorder; Goals; Harvest; Hypersensitivity; In Vitro; Injury; Instruction; Knock-out; Knockout Mice; Knowledge; Laboratories; Lumbar spinal cord structure; Maintenance; Measures; Mechanics; Mediating; Mediator of activation protein; Microglia; Migraine; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; Nerve; Nervous system structure; Neuraxis; Neuroglia; Neurons; Neurotransmitters; Nitric Oxide; Nociceptors; Pain; Pathway interactions; Peptides; Peripheral Nerves; Peripheral nerve injury; Persistent pain; Phase; Play; Primary Lesion; Production; Reporting; Rodent; Role; Sensory; Signal Pathway; Signal Transduction; Signaling Molecule; Small Inducible Cytokine A3; Spinal; Spinal Cord; Spinal Ganglia; Spinal cord injury; Spinal nerve structure; Structure of trigeminal ganglion; Surface; TNFRSF5 gene; Tactile; Testing; Tumor Necrosis Factor Receptor; Up-Regulation; Western Blotting; Wild Type Mouse; base; chemokine; chronic pain; design; drug development; effective therapy; human NOS2A protein; in vivo; nerve injury; neutralizing antibody; novel; pain behavior; painful neuropathy; preclinical study; receptor; relating to nervous system; response

Decades of studies have provided ample evidence that glial neuroimmune activation contributes in the development of neuropathic pain. However, knowledge regarding the role of neural-glial interactions in the pathophysiology of neuropathic pain is currently lacking. In preclinical studies, calcitonin gene related peptide (CGRP), a peptide neurotransmitter, has been shown to be involved in peripheral nerve injuryinduced tactile hypersensitivity, a behavioral sign of neuropathic pain. Previously, we have also demonstrated a critical role of central nervous system (CNS) microglial CD40 in the maintenance of mechanical hypersensitivity post-peripheral nerve injury. In the current study, the possible interactions between microglial CD40 signaling and primary afferent neuron-released CGRP in regards to they affect glial cells in the spinal cord will be investigated. We hypothesize that following peripheral nerve injury, spinal cord CD40 mediates the release of CGRP from primary afferent nociceptors and this CGRP release promotes persistent pain behavior, particularly during the maintenance phase, through the induction of proinflammatory chemokine production by glia. This central hypothesis will be tested via both in vivo and in vitro studies. An established rodent neuropathic pain model, spinal nerve L5 transection (L5Tx), will be used for the in vivo studies. Dorsal root ganglia (DRG) neuron-microglia co-cultures and mixed glial cultures will be generated for the in vitro studies. The central hypothesis will be tested through 4 specific aims: 1) Examine lumbar spinal cord chemokine production following L5Tx in CD40 KO mice vs. WT mice and the role of selected chemokines in the development of behavioral hypersensitivity; 2) Determine whether CGRP is involved in L5Tx-induced chemokine production and behavioral hypersensitivity; 3) Evaluate the role of microglial CD40 in CGRP release by primary afferent neurons; and 4) Assess glial production of selected proinflammatory chemokines and upstream signaling pathways (mainly mitogen-activated protein kinase pathways) following CGRP stimulation in mixed glial cultures. The long-term goal of our study is to further understand the pathophysiology of nerve injury-induced neuropathic pain in order to uncover novel targets for new drug development leading to more efficacious treatments of neuropathic pain.

数十年的研究提供了充分的证据，表明神经胶质神经免疫性激活有助于 神经性疼痛的发展。但是，关于神经互动在 目前缺乏神经性疼痛的病理生理。在临床前研究中，降钙素基因相关 肽（CGRP）是一种肽神经递质，已证明与周围神经损伤有关 触觉超敏反应，是神经性疼痛的行为标志。以前，我们也有 表现出中枢神经系统（CNS）小胶质CD40在维持中的关键作用 机械性超敏反应后神经损伤。在当前的研究中，可能的相互作用 在小胶质细胞CD40信号传导和主要传入神经元释放的CGRP之间 将研究脊髓中的神经胶质细胞。我们假设在周围神经损伤之后， 脊髓CD40介导了从初级传入的伤害感受器中释放CGRP，并且该CGRP 释放促进持续性疼痛行为，尤其是在维护阶段，通过 神经胶质诱导促炎性趋化因子的产生。该中心假设将通过 体内和体外研究。已建立的啮齿动物神经性疼痛模型，脊神经L5横断 （L5TX）将用于体内研究。背根神经节（DRG）神经元 - 神经元共培养和 用于体外研究将产生混合的神经胶质培养物。中心假设将通过4进行检验 具体目的：1）在CD40 KO小鼠VS中L5TX后检查腰脊髓趋化因子的产生。 WT小鼠和选定趋化因子在行为超敏反应发展中的作用； 2） 确定CGRP是否参与L5TX诱导的趋化因子产生和行为 超敏反应； 3）评估小胶质细胞CD40在原发性传入神经元释放CGRP中的作用；和 4）评估选定促炎性趋化因子和上游信号通路的神经胶质产生 （主要是促有丝分裂原激活的蛋白激酶途径）在混合神经胶质培养物中刺激后。这 我们研究的长期目标是进一步了解神经损伤引起的神经性的病理生理学 疼痛以发现新的药物开发的新目标，从而更有效地治疗 神经性疼痛。