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

项目总结/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重大空白 以及它们参与初级传入敏化。对周围神经胶质细胞的作用缺乏了解 细胞，主要是由于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， 在体内，提出了一个重要的障碍，在发展创造性的和有效的策略，研究胶质细胞 对慢性病的贡献。该项目的长期目标是针对GFAP+卫星胶质细胞（SGCs） 用于预防和治疗外周致敏的基因疗法。这项建议的目的是 识别感觉神经节中SGC-神经元相互作用的信号通路，以及 表征参与内脏疼痛和膀胱炎的感觉SGC信号传导的翻译组变化 功能障碍强有力的初步数据和过去的发现，在感觉SGCs的背景下，慢性疼痛导致 核心假设是SGCs中的Gq-GPCR信号传导有效地降低了膀胱的兴奋性和活动性， 投射感觉神经元并有助于膀胱功能的神经控制。这一假设将是 通过追求三个具体目标进行测试：1）测试感觉卫星神经胶质细胞Gq-GPCR信号传导的假设， 在生理条件和炎症中降低膀胱传入敏感性并改变排尿- 诱导的内脏疼痛和膀胱过度活动模型; 2）确定卫星胶质细胞Gq-1和Gq-2之间的分子联系。 GPCR激活和感觉神经元兴奋性降低;和3）发现感觉卫星的变化 炎症性盆腔疼痛和下尿路感染发病和慢性期胶质细胞翻译蛋白的表达 上消化道症状（LUTS）。在第一个目标下，外周GFAP+胶质细胞中的靶向和选择性基因操作 研究SGCs在膀胱传入神经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 在感觉神经节外植体中使用类似于研究CNS神经胶质-神经元的方法进行鉴定 现场互动。将在体内进行额外的实验，以测试靶向的治疗潜力。 在此目的中，鉴定的用于减轻内脏疼痛和膀胱过度活动的信号传导途径。下 第三个目标，TRAP技术将被用来识别感觉SGC翻译组的变化， 炎症引起的膀胱过度活动和盆腔疼痛。拟议的研究是 意义重大，因为1）它证明了一种选择性激活外周血淋巴细胞的新的药物遗传学方法， 神经胶质细胞在体内，这有利于更广泛的研究社区在神经泌尿学研究; 2）它有望填补 关于膀胱功能的神经胶质调节的知识差距，这是一个完全未开发的研究领域， 治疗潜力; 3）它为后续的临床前和临床试验提供了一线证据， 靶向卫星神经胶质细胞和外周腺苷受体治疗盆腔疼痛; 4）它奠定了 为生理和疾病期间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.