Cellular mechanisms of antidepressant drug actions in neuropathic pain models

项目总结 神经性疼痛是一种以感觉、认知和情感症状为特征的慢性疾病。大部分 用于治疗这种疾病的药物疗效低，副作用大。了解 调控外周血细胞传递和维持的表观遗传和转录机制 神经损伤(PNI)症状将是发展新的治疗方法的重要一步。我们的 早期的研究发现，表观遗传修饰物组蛋白脱乙酰酶5(HDAC5)在急性髓细胞白血病的发病中起关键作用。 抗抑郁药在神经病理性疼痛模型中的作用和疗效。伏核(NAC)中的HDAC5 结合染色质复合体以抑制影响突触功能的几个基因的表达， 包括转录因子MEF2C。父R01提案调查了 HDAC5及其下游靶点调节NAC中抗抑郁药的作用。本补充项目 旨在阐明细胞类型特异性的表观遗传学和转录学机制介导的抗痛觉过敏 单胺类抗抑郁药在神经病理性疼痛模型中的作用。单细胞生物信息学分析 核RNAseq数据将补充关于HDAC5和MEF2C行动的发现，并将提供关于 与抗抑郁疗效相关的转录事件。我们将使用荧光技术验证关键发现 原位杂交、免疫荧光以及蛋白质印迹分析。药理和遗传模型 将用于测试选定的基因/细胞内途径。对调节分子机制的理解 单胺类抗抑郁药物的作用将有助于开发新的更有效的抗抑郁药物 治疗神经性疼痛的药物。

PROJECT SUMMARY Neuropathic pain is a chronic condition characterized by sensory, cognitive and affective symptoms. Most of the drugs used to treat this disorder demonstrate low efficacy and major side-effects. Understanding the epigenetic and transcriptional mechanisms that modulate the transmission and maintenance of peripheral nerve injury (PNI) symptoms will be a major step towards the development of novel treatment approaches. Our earlier findings identified a key role of the epigenetic modifier Histone deacetylase 5 (HDAC5) in the onset of action and efficacy of antidepressants in models of neuropathic pain. HDAC5 in the Nucleus Accumbens (NAc) binds to chromatin complexes to suppress the expression of several genes that affect synaptic function, including the transcription factor MEF2C. The parent R01 proposal investigates the mechanism by which HDAC5 and downstream targets modulate the actions of antidepressants in the NAc. This supplement project aims to elucidate cell type-specific epigenetic and transcriptomic mechanisms mediating the antiallodynic effects of monoamine-targeting antidepressants in models of neuropathic pain. Bioinformatic analysis of single nuclei RNASeq data will complement findings on HDAC5 and MEF2C actions and will also provide insight on transcriptomic events associated with antidepressant efficacy. We will validate key findings using fluorescent in situ hybridization, immunofluorescence, as well as western blot analysis. Pharmacological and genetic models will be use to test selected genes/intracellular pathways. Understanding the molecular mechanisms mediating the actions of monoamine-targeting antidepressants will help the development of novel and more efficacious medications for the treatment of neuropathic pain.