Project 1: Cao

项目一：曹

• 批准号: 8529580
• 负责人: Ling Cao
• 金额: $ 20.04万
• 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
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8529580
• 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，神经胶质免疫激活在 神经病理性疼痛的发展。然而，关于神经-神经胶质相互作用在 神经病理性疼痛的病理生理学目前尚不清楚。在临床前研究中，降钙素基因相关 降钙素基因相关肽(CGRP)是一种多肽类神经递质，参与周围神经损伤。 触觉过敏，神经病理性疼痛的行为征兆。此前，我们还 证实中枢神经系统(CNS)小胶质细胞CD40在维持性脑出血中的关键作用 周围神经损伤后的机械超敏反应。在目前的研究中，可能的相互作用 小胶质细胞CD40信号与初级传入神经元释放的降钙素基因相关肽之间的关系 将对脊髓中的神经胶质细胞进行研究。我们假设在周围神经损伤后， 脊髓CD40介导初级传入伤害性感受器CGRP的释放 释放促进持续的疼痛行为，特别是在维护阶段，通过 胶质细胞诱导炎性趋化因子的产生。这一中心假设将通过以下方式进行检验 包括体内和体外研究。大鼠神经病理性疼痛模型的建立--L5脊神经横断 (L5Tx)，将用于体内研究。背根神经节(DRG)神经元-小胶质细胞共培养和 将为体外研究产生混合的胶质细胞培养物。核心假设将通过4个阶段进行检验 具体目的：1)检测CD40 KO小鼠与小鼠L5Tx后腰髓趋化因子的产生。 WT小鼠及部分趋化因子在行为超敏反应中的作用 确定CGRP是否参与L5Tx诱导的趋化因子的产生和行为 3)评价小胶质细胞CD40在初级传入神经元释放CGRP中的作用； 4)评估选定的促炎趋化因子和上游信号通路的胶质细胞产生 (主要是有丝分裂原激活的蛋白激酶通路)在混合神经胶质细胞培养中CGRP刺激后。这个 我们研究的长期目标是进一步了解神经损伤引起的神经病的病理生理学。 疼痛，以发现新药开发的新靶点，从而导致更有效的治疗 神经性疼痛。