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.

摘要 炎性和抗炎性细胞因子参与痛觉传递中的神经元超兴奋性 小路。白介素1(IL-1)是一种炎症细胞因子，参与多种神经免疫反应。 特别是，已经证明IL-1诱导急性疼痛，降低吗啡的止痛活性，是 参与耐受性的形成，是慢性神经性和炎症性疼痛所必需的。这有 已在许多方面进行了研究，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。从机械的观点来看，这些实验的问题是IL-1受体 内源性存在于大脑中的各种细胞类型上，包括星形胶质细胞、小胶质细胞、内皮细胞、 和神经元，而全球基因敲除实验不能定义介导IL-1活动的细胞类型。这个 发展新的遗传模型，在这种模型中，IL-1受体IL-1R1可以相互敲除并 从全局基因敲除恢复(IL-1R1r/r)到脑、脊髓和背根的单个细胞类型 神经节可以更具体地识别IL-1在疼痛方面的作用。具体目标1 将研究5种小鼠的吗啡镇痛和耐受性发展，野生型，全局IL-1R1KO， 以及有选择地将IL-1R1受体恢复到神经元、内皮细胞和星形胶质细胞的小鼠。我们 预计将发现全球KO动物和两个恢复的小鼠基因型将具有更强的 如文献所述，吗啡止痛时间延长，耐受性降低。此外， 其中一个恢复的小鼠品系将表现得像野生型动物，吗啡活性降低，并将展示 宽容发展。Aim 2将研究两种慢性疼痛模型，脊髓神经结扎术和完全 弗氏佐剂，使用相同的小鼠基因型别来确定哪种细胞类型参与了发育 慢性疼痛的症状。Aim 3将IL-1R1r/r小鼠与c-Fos-Cre/ERT2杂交，建立一种新的小鼠模型 (TRAP2)小鼠。有了这种新的遗传模型，IL-1R1将只在受到 一些疼痛刺激。使用这些新的遗传模型将确定IL-1与阿片类药物有关的作用 止痛和疼痛到特定类型的细胞，并发展一种合作，将能够确定 IL-1的作用机制，在未来的R01应用中。