Activating Peripheral Glia to Relieve Visceral Pain in Animal Models of Urological Chronic Pelvic Pain Syndrome (UCPPS)

项目摘要/摘要 我们对外周GFAP胶质细胞如何调节感觉神经元活动的认识存在显著差距。 以及它们参与初级传入敏感化。对外周神经胶质细胞的作用缺乏了解 细胞，主要是因为我们不能选择性地干扰周围神经胶质细胞亚群中的信号通路 在活体内，是开发创造性和有效的研究神经胶质细胞的策略的重要障碍 在慢性病方面的贡献。这个项目的长期目标是瞄准GFAP卫星胶质细胞(SGCs)。 用于预防和治疗外周敏感症的基因疗法。这项建议的目的是 确定感觉神经节中SGC-神经元相互作用的信号通路，以及 内脏痛和膀胱中感觉SGC信号的翻译组变化特征 功能障碍。在慢性疼痛的背景下，强烈的初步数据和感觉性SGCs的过去发现导致了 中心假设GQ-GPCR信号在SGCs中有效地降低了膀胱的兴奋性和活性- 投射感觉神经元，有助于神经控制膀胱功能。这一假设将是 通过追求三个特定的目标进行检验：1)检验感觉卫星Gq-GPCR信号的假设 降低膀胱传入敏感度，改变生理状态和炎症状态下的排尿- 诱导内脏痛和膀胱过度活动模型；2)确定卫星神经胶质细胞GQ-2之间的分子联系。 GPCR激活和感觉神经元兴奋性降低；3)发现感觉卫星的变化 炎症性盆腔疼痛和下尿路疼痛发病和慢性期的胶质细胞翻译组 肠道症状(LUTS)。在第一个目标下，外周GFAP胶质细胞的靶向和选择性基因操作 将用于研究SGCs在膀胱传入的生理和病理调节中的作用 兴奋性。在第二个目标中，SGC-神经元相互作用的细胞和分子机制将 用类似于研究中枢神经系统神经胶质细胞的方法在感觉神经节外植体中鉴定 原位相互作用。更多的实验将在体内进行，以测试靶向治疗的潜力。 已确定的减轻内脏疼痛和膀胱过度活动的信号通路(S)。在.之下 第三个目标，TRAP技术将被用来确定感觉SGC翻译组的变化作为 炎症引起的膀胱过度活动和盆腔疼痛是一种无偏见的方法。 之所以重要，是因为1)它展示了一种选择性激活外周血细胞的新药物遗传学方法 体内的胶质细胞，这将使神经病学研究中更广泛的研究领域受益；2)有望填补 关于膀胱功能的神经胶质调节的知识鸿沟--一个完全未被探索的研究领域 治疗潜力；3)它为随后的临床前和临床试验提供了第一线证据 靶向卫星神经胶质细胞和外周腺苷受体治疗盆腔疼痛；4)奠定了 为生理和疾病过程中外周神经胶质细胞特定亚群的遗传分析奠定基础。

PROJECT SUMMARY/ABSTRACT There are significant gaps in our knowledge of how peripheral GFAP+ glia regulate sensory neuronal activity and their involvement in primary afferent sensitization. The lack of understanding of the roles of peripheral glial cells, mainly due to our inability of selectively perturbing signaling pathways in subpopulations of peripheral glia in vivo, presents an important obstacle in developing creative and effective strategies for investigating glial contribution in chronic diseases. The long-term goal of this project is to target GFAP+ satellite glial cells (SGCs) for gene therapies for preventing and treating peripheral sensitization. The objective of this proposal is to identify signaling pathways underlying SGC-neuron interaction in the sensory ganglia, as well as to characterize the translatome changes in sensory SGC signaling involved in visceral pain and bladder dysfunction. Strong preliminary data and past findings in sensory SGCs in the context of chronic pain led to the central hypothesis that Gq-GPCR signaling in SGCs potently decreases the excitability and activity in bladder- projecting sensory neurons and contributes to neural control of bladder functions. This hypothesis will be tested by pursuing three specific aims: 1) Test the hypothesis that sensory satellite glial Gq-GPCR signaling decreases bladder afferent sensitivity and alters micturition in physiological conditions and in inflammation- induced visceral pain and bladder overactivity model; 2) Identify the molecular link between satellite glial Gq- GPCR activation and decreased sensory neuronal excitability; and 3) Discover the changes in sensory satellite glial translatome during the onset and chronic phase of inflammation-induced pelvic pain and lower urinary tract symptoms (LUTS). Under the first aim, targeted and selective gene manipulation in peripheral GFAP+ glia will be utilized to study the role of SGCs in physiological and pathological regulation of bladder afferent excitability. In the second aim, the cellular and molecular mechanism underlying SGC-neuron interaction will be identified in sensory ganglia explants using approaches similar to those used in studying CNS glial-neuronal interactions in situ. Additional experiments will be performed in vivo to test the therapeutic potential of targeting the identified signaling pathway(s) for alleviating visceral pain and bladder overactivity in this aim. Under the third aim, TRAP technology will be employed to identify changes in sensory SGC translatome as a function of inflammation-induced bladder overactivity and pelvic painin a non-biased manner.The proposed research is significant because 1) it demonstrates a new pharmacogenetic approach for selectively activating peripheral glia in vivo, which benefits broader research community in Neuro-urology research; 2) it is expected to fill the knowledge gap on glial modulation of bladder function, a completely unexplored research field with high therapeutic potential; 3) it provides the first line of evidence for subsequent pre-clinical and clinical trials of targeting satellite glial cells and peripheral adenosine receptors in treating pelvic pain; and 4) it lays the groundwork for genetic analysis on specific subsets of peripheral glia during physiology and disease.