Cell Specific Identification of IL-1R Actions in Acute and Chronic Pain

IL-1R 在急性和慢性疼痛中作用的细胞特异性鉴定

• 批准号: 10317169
• 负责人: Ning Quan
• 金额: $ 41.11万
• 依托单位: FLORIDA ATLANTIC UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10317169
• 关键词: AVIL gene; Absence of pain sensation; Acute Pain; Afferent Neurons; Analgesics; Animals; Anti-Inflammatory Agents; Astrocytes; Attenuated; Behavioral; Biochemical; Brain; Brain region; Breeding; Cells; Collaborations; Cre driver; Development; Endothelial Cells; Enterobacteria phage P1 Cre recombinase; Exhibits; FOS gene; Freund' s Adjuvant; Future; Genetic; Genetic Models; Genetic Recombination; Genotype; Grant; Individual; Inflammatory; Interleukin-1; Interleukin-1 Receptors; Interleukin-1 beta; Knock-out; Knockout Mice; Lead; Ligation; Literature; Location; Maintenance; Mechanics; Mediating; Microglia; Modality; Modeling; Morphine; Mus; Neuroglia; Neuroimmune; Neurons; Opioid; Opioid Analgesics; Pain; Pathway interactions; Population; Receptor Cell; Receptor Signaling; Resistance; Signal Transduction; Site; Spinal Cord; Spinal Ganglia; Spinal nerve structure; Stimulus; Testing; Visualization; allodynia; anakinra; brain endothelial cell; cell type; chronic neuropathic pain; chronic pain; conditional knockout; cytokine; experimental study; genetic manipulation; inflammatory pain; interleukin-1 receptor type I; mouse genetics; mouse model; neuroimmunology; neuroinflammation; novel; pain behavior; pain model; painful neuropathy; receptor; receptor expression; response; restoration; tool; transmission process

Abstract Inflammatory and anti-inflammatory cytokines contribute to neuronal hyperexcitability in pain transmission pathways. One of the inflammatory cytokines, Interleukin-1 (IL-1), is involved in many neuroimmune responses. In particular, it has been demonstrated that IL-1 induces acute pain, reduces morphine analgesic activity, is involved in tolerance development, and is necessary for chronic neuropathic and inflammatory pain. This has been studied in many ways, including use of an endogenous IL-1 receptor antagonist, and the use of IL-1 and IL-1R knockout mice. The problem with these experiments from a mechanistic standpoint is that IL-1 receptors are found endogenously on a variety of cell types in the brain, including astrocytes, microglia, endothelial cells, and neurons, and global knockout experiments can't define the cell types that mediate the actions of IL-1. The development of novel genetic models, in which the IL-1 receptor, IL-1R1, can be reciprocally knocked out and restored (IL-1R1r/r) from a global knockout, into individual cell types in the brain, spinal cord, and dorsal root ganglion permits a much more specific identification of the actions of IL-1 with respect to pain. Specific Aim 1 will study morphine analgesia and tolerance development in 5 mouse genotypes, wild type, global IL-1R1 KO, and mice in which the IL-1R1 receptor is restored selectively into neurons, endothelial cells, and astrocytes. We expect to find that the global KO animals and two of the restored mouse genotypes will have more potent and prolonged morphine analgesia and reduced tolerance development, as described in the literature. Furthermore, one of the restored mouse lines will act like the wild type animals with reduced morphine activity and will exhibit tolerance development. Aim 2 will examine two chronic pain models, spinal nerve ligation and Complete Freund's Adjuvant, using the same mouse genotypes, to determine which cell type mediates the development of chronic pain. Aim 3 will develop a new mouse model by crossing the IL-1R1r/r mice with c-Fos-Cre/ERT2 (TRAP2) mice. With this new genetic model, IL-1R1 will be restored only in mice that have been subjected to some pain stimulus. The use of these novel genetic models will pinpoint the actions of IL-1 with respect to opioid analgesia and pain to specific cell types and develop a collaboration that will be able to determine the mechanisms of IL-1 actions, in future R01 applications.

抽象的 炎症和抗炎细胞因子导致疼痛传递中的神经元过度兴奋 途径。白细胞介素-1 (IL-1) 是炎症细胞因子之一，参与许多神经免疫反应。 特别是，已证明IL-1可诱发急性疼痛，降低吗啡镇痛活性， 参与耐受性发展，对于慢性神经性疼痛和炎症性疼痛是必需的。这有 已通过多种方式进行了研究，包括使用内源性 IL-1 受体拮抗剂，以及使用 IL-1 和 IL-1R 基因敲除小鼠。从机制的角度来看，这些实验的问题在于 IL-1 受体 内源性存在于大脑中的多种细胞类型中，包括星形胶质细胞、小胶质细胞、内皮细胞、 和神经元，并且全局敲除实验无法定义介导 IL-1 作用的细胞类型。这 开发新的遗传模型，其中 IL-1 受体 IL-1R1 可以相互敲除， 从整体敲除中恢复（IL-1R1r/r）到大脑、脊髓和背根中的单个细胞类型 神经节允许更具体地识别 IL-1 对疼痛的作用。具体目标 1 将研究 5 种小鼠基因型、野生型、整体 IL-1R1 KO 的吗啡镇痛和耐受性发展， 小鼠的 IL-1R1 受体选择性地恢复到神经元、内皮细胞和星形胶质细胞中。我们 期望发现全球KO动物和两种恢复的小鼠基因型将具有更有效和更有效的基因型。 如文献中所述，延长吗啡镇痛时间并降低耐受性的发展。此外， 恢复的小鼠品系之一将表现得像野生型动物一样，但吗啡活性降低，并且会表现出 耐受性的发展。目标 2 将检查两种慢性疼痛模型：脊神经结扎和完全 弗氏佐剂，使用相同的小鼠基因型，以确定哪种细胞类型介导发育 慢性疼痛。 Aim 3 将通过将 IL-1R1r/r 小鼠与 c-Fos-Cre/ERT2 杂交来开发新的小鼠模型 (TRAP2) 小鼠。通过这种新的遗传模型，IL-1R1 只会在已经接受过治疗的小鼠中恢复。 一些疼痛刺激。这些新型遗传模型的使用将查明 IL-1 对阿片类药物的作用 针对特定细胞类型的镇痛和疼痛，并开展合作来确定 IL-1作用机制，在未来的R01应用中。