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: 10259799
• 负责人: Michael P Jankowski
• 金额: $ 38.56万
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10259799
• 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; repair function; 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：外围损伤反应需要目标组织，免疫细胞和 主要感觉神经元。这些系统之间的串扰对于伤害后的肌肉修复至关重要 伤害感受。尽管了解免疫系统在功能恢复中的作用有很多了解 在肌肉和疼痛发育中，尚不清楚循环免疫细胞与肌纤维的物理偶联是否存在 损伤后神经元直接调节这两个独特的生物学过程。有关小说的信息 肌肉，周围神经系统和免疫细胞之间的相互作用可以显着前进 了解肌痛和肌肉修复。这项研究的目的是确定是否浸润免疫细胞 受伤后，电气夫妇与肌纤维或神经元进行双重调节功能性肌肉修复和 伤害感受。最近的报告表明，在受伤后，连接蛋白43（CX43）的间隙连接可能形成 巨噬细胞和肌纤维调节修复。还显示出类似的间隙连接形式 背根神经节（DRG）内的相邻神经元之间以调节伤害感受。在心中 驻留巨噬细胞和心肌细胞之间的电耦合对于适当的房室至关重要 传导。因此，合理地假设免疫细胞将免疫细胞与肌纤维搭配在一起，并且 骨骼肌损伤后的伤害感受器，以协调对损伤的反应并进行双重调节组织修复 和痛苦。 AIM 1（R61相）将确定巨噬细胞向伤害感受器的电耦合是否会调节 切口相关的超敏反应。这项研究将使用新颖的转基因策略来专门敲除CX43 在带后爪肌肉切口的小鼠的巨噬细胞中。这将与化学发生或 巨噬细胞的声遗传激活。 CX43敲除和巨噬细胞激活的影响将是 通过我们的离体肌肉传入记录准备，与肌肉疼痛相关的行为测试和 在巨噬细胞和原代DRG细胞共培养中的钙成像（使用GCAMP6记者）。目标2 （R61相）将使用类似的组来确定渗透巨噬细胞的电耦合是否与肌纤维 切口后促进肌肉组织的修复。这些操作的影响将使用 后爪肌肉细胞的钙成像，肌电图/复合肌肉动作潜在记录 体内和肌肉膜完整性的解剖学分析。 AIM 3（R33阶段）将进一步探索 使用不同的转基因组合，抑制性化学遗传学的巨噬细胞电耦合的功能 以及更严重的肌肉损伤模型。结果将允许确定新颖的交流方式 在循环中的免疫因子和它们正在影响的周围结构之间。数据将提供新颖 对肌肉损伤反应的见解将远远超出当前报告的增量扩张。 这些见解可以识别出许多用于疼痛或肌肉修复治疗干预的新目标 肌肉损伤状态。