Electrical Coupling of Circulating Immune Cells to Peripheral Tissues

循环免疫细胞与周围组织的电耦合

• 批准号: 10078364
• 负责人: Michael P Jankowski
• 金额: $ 41.34万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-08 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078364
• 关键词: Action Potentials; Affect; Afferent Neurons; Anatomy; Animals; Basal lamina; Behavior assessment; Biological Assay; Biological Process; CHRM3 gene; Calcium; Cardiac Myocytes; Cells; Coculture Techniques; Communication; Complex; Connexin 43; Coupling; Data; Development; Dyes; Electromyography; Gap Junctions; Genetic; Goals; Heart; Hypersensitivity; Image; Immune; Immune response; Immune system; Immunologic Factors; Injury; Investigation; Ion Channel Gating; Knock-out; Macrophage Activation; Membrane; Modeling; Mus; Muscle; Muscle Cells; Muscle Fibers; Muscle function; Myalgia; Myopathy; Nerve; Neurons; Nociception; Nociceptors; Organism; Pain; Pain management; Peripheral; Peripheral Nervous System; Phase; Play; Population; Positioning Attribute; Preparation; Process; Recovery; Recovery of Function; Reporter; Reporting; Research; Role; Shapes; Signal Transduction; Skeletal Muscle; Spinal Cord; Spinal Ganglia; Structure; Surgical incisions; System; Techniques; Testing; Therapeutic Intervention; Time; Tissues; Transgenic Mice; Transgenic Organisms; Tumor-infiltrating immune cells; Ultrasonography; Ventricular; behavior test; cell type; chemokine; cytokine; designer receptors exclusively activated by designer drugs; functional restoration; ganglion cell; in vitro Assay; in vivo; insight; knowledge of results; macrophage; multidisciplinary; myogenesis; new therapeutic target; nociceptive response; novel; repaired; response; response to injury; restoration; tissue repair

Abstract: Peripheral injury responses require sophisticated interactions of target tissues, immune cells and primary sensory neurons. Crosstalk between these systems is essential for post-injury muscle repair and nociception. While a great deal is known about the role of the immune system in functional restoration of muscles and in pain development, it is not known if physical coupling of circulating immune cells to myofibers or neurons after injury directly modulates both of these unique biological processes. Information on novel interactions between muscles, the peripheral nervous system and immune cells could significantly advance understanding of myalgia and muscle repair. The goal of this study is to determine if infiltrating immune cells electrically couple to myofibers or neurons after injury to dually modulate functional muscle repair and nociception. Recent reports suggest that, after injury, connexin 43 (Cx43) gap junctions may form between macrophages and myofibers to modulate repair. It has also been shown that similar gap junctions form between adjacent neurons within the dorsal root ganglion (DRG) to regulate nociception. In the heart, electrical coupling between resident macrophages and cardiomyocytes is crucial for proper atrioventricular conduction. It is therefore reasonable to hypothesize that immune cells electrically couple to myofibers and nociceptors after skeletal muscle damage to coordinate responses to injury and dually modulate tissue repair and pain. Aim 1 (R61 Phase) will determine if electrical coupling of macrophages to nociceptors modulates incision-related hypersensitivity. This study will use novel transgenic strategies to specifically knockout Cx43 in macrophages in mice with hind paw muscle incision. This will be used in conjunction with chemogenetic or sono-genetic activation of macrophages. Impact of Cx43 knockout and macrophage activation will be assessed with our ex vivo muscle afferent recording preparations, muscle pain-related behavioral tests and calcium imaging (using GCaMP6 reporters) in co-cultures of macrophages and primary DRG cells. Aim 2 (R61 Phase) will use similar groups to determine if electrical coupling of infiltrating macrophages to myofibers facilitates repair of muscle tissue after incision. The impact of these manipulations will be determined using calcium imaging of hind paw muscle cells, electromyography/ compound muscle action potential recordings in vivo, and anatomical analyses of muscle membrane integrity. Aim 3 (R33 Phase) will further explore the functions of macrophage electrical coupling using different transgenic combinations, inhibitory chemogenetics and more severe models of muscle injury. Results will allow determination of novel means of communication between circulating immune factors and the peripheral structures they are affecting. Data will provide novel insights into muscle injury responses that will go well beyond the incremental expansion of current reports. These insights could identify a novel target for therapeutic intervention for pain or muscle repair in numerous muscle injury states.

摘要：外周损伤反应需要靶组织、免疫细胞和 初级感觉神经元。这些系统之间的串扰对于损伤后肌肉修复和 伤害性感受。虽然很多人都知道免疫系统在心脏功能恢复中的作用 在肌肉和疼痛的发展中，循环免疫细胞是否与肌纤维发生物理偶联尚不清楚 或损伤后的神经元直接调节这两个独特的生物过程。关于小说的信息 肌肉、外周神经系统和免疫细胞之间的相互作用可能会显著促进 了解肌肉疼痛和肌肉修复。这项研究的目标是确定渗入的免疫细胞 损伤后电耦合到肌纤维或神经元，以双重调节功能性肌肉修复和 伤害性感受。最近的报道表明，损伤后，连接蛋白43(Cx43)的缝隙连接可能在 巨噬细胞和肌纤维调节修复。也有研究表明，类似的缝隙连接形成 在背根神经节(DRG)内相邻神经元之间调节伤害性感觉。在内心深处， 驻留巨噬细胞和心肌细胞之间的电偶联是房室正常的关键 传导。因此，可以合理地假设免疫细胞与肌纤维电偶联 骨骼肌损伤后的伤害性感受器协调损伤反应和双重调节组织修复 和痛苦。目标1(R61阶段)将确定巨噬细胞与伤害性感受器的电耦合是否调制 与手术有关的过敏症。这项研究将使用新的转基因策略来特异性地敲除Cx43 在后爪肌切开的小鼠巨噬细胞中。这将与化学发生或联合使用 巨噬细胞的声学激活。Cx43基因敲除和巨噬细胞激活的影响将是 通过我们的体外肌肉传入记录准备、肌肉疼痛相关的行为测试和 巨噬细胞和原代DRG细胞共培养中的钙成像(使用GCaMP6报告)。目标2 (R61阶段)将使用类似的小组来确定浸润性巨噬细胞与肌纤维的电偶联 促进切开后肌肉组织的修复。这些操作的影响将通过以下方式确定 大鼠后爪肌细胞钙显象、肌电/复合肌肉动作电位记录 活体，以及对肌膜完整性的解剖分析。目标3(R33阶段)将进一步探索 不同转基因组合、抑制性化学遗传学对巨噬细胞电偶联功能的影响 以及更严重的肌肉损伤模型。结果将使确定新的通信手段成为可能 循环免疫因子和它们所影响的外周结构之间的关系。数据将提供新颖的 对肌肉损伤反应的洞察将远远超出当前报告的增量扩展。 这些洞察力可以为治疗疼痛或肌肉修复确定一个新的靶点 肌肉受伤的状态。