Glycine Receptor Expression in Sensory Afferents to Modulate Pain Signaling

感觉传入中甘氨酸受体的表达调节疼痛信号传导

基本信息

批准号：
8520405
负责人：
Joseph C Glorioso
金额：
$ 34.8万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8520405
关键词：
Absence of pain sensation Address Adverse effects Affect Afferent Neurons American Analgesics Animal Model Antibodies Behavioral Assay C Fiber Cell Surface Receptors Complement Development Disease Elements Engineering Enhancers Fiber Functional disorder Future Gene Delivery Gene Expression Gene Transfer Glycine Glycine Receptors Glycoproteins Goals Health Heating Herpesvirus 1 Hypersensitivity Immunohistochemistry Infection Inflammation Injection of therapeutic agent Injury Lead Ligands Maintenance Maps Mechanics Mediating Medical Methods Modeling Nerve Fibers Neurons Nociceptors Pain Pain management Peripheral Persistent pain Pharmacotherapy Phenotype Physiology Population Proprioception Rattus Risk Role Sensory Series Signal Transduction Specificity Spinal Ganglia Staging Stimulus Technology Testing Time Tissues Touch sensation Transgenes Viral Viral Vector Virus Latency afferent nerve base chronic pain design gene therapy gene transfer vector in vivo inflammatory neuropathic pain inflammatory pain insight nerve injury novel painful neuropathy pressure promoter receptor receptor expression research study response selective expression success targeted delivery transgene expression vector

项目摘要

DESCRIPTION (provided by applicant): Chronic pain is a major health concern affecting 80 million Americans at some time in their lives. Current pharmacotherapies are not effective long-term, which has led to the development and testing of gene therapy approaches. We and others have demonstrated that herpes simplex virus type 1 (HSV) based vectors can deliver highly effective pain-modulating transgenes to sensory neurons in vivo following inoculation of peripheral tissue. One major advantage of this approach is that painful tissue can be specifically targeted by local vector delivery. We believe that this advantage can be further extended by targeting specific neuron types responsible for chronic pain, thus enabling gene transfer to be tailored to specific types of pain, such as inflammatory or neuropathic pain, while simultaneously reducing deleterious side effects. We have recently shown that HSV-mediated transfer and long-term expression of the glycine receptor 11 subunit (GlyR11) can be used to control the timing and duration of afferent silencing with exogenous administration of glycine. Based on these findings and our recent success in the creation of highly efficient, fully retargeted HSV vectors, we hypothesize that we can selectively silence distinct subpopulations of primary afferents responsible for neuropathic and inflammatory pain, therefore providing injury specific pain relief. These studies will enable us to determine whether the same or different afferent populations underlie inflammatory and neuropathic pain and provide the rational basis for the future development of HSV-based gene transfer vectors designed to restrict analgesia to the relevant primary afferents. We anticipate that these studies will (i) provide a novel platform technology that will allow us to selectively express transgenes designed to modulate the function of sensory afferent subpopulations, a strategy that can be readily extended to other types of sensory nerve disorders; (ii) develop initial functional and physical maps of sensory afferent subtypes that upon silencing will block different persistent pain conditions, thereby providing essential information needed for targeted pain control; (iii) identify afferents that have been functionally altered to respond to painful stimuli providing further information on nerve fiber plasticity; and, (iv) identify potential risks associated with silencing of an inappropriate population of sensory neurons (e.g. altered proprioception). To achieve these goals, we have proposed a series of interrelated experiments described in 3 Specific Aims. In Aim 1, the infectivity of HSV vectors will be retargeted to selectively transduce (a) A2 fibers, (b) peptidergic and (c) nonpeptidergic C fibers. In Aim 2, we will complement transductional retargeting with transcriptional targeting using transgene promoters that will, when combined with transductional targeting, fine tune silencing specificity. The combination strategy is intended to create initial fine maps of subpopulations of sensory fibers within the larger transductionally targeted groups representing critical afferents for the response to different painful stimuli. In Aim 3, the retargeted HSV vectors will be introduced into the DRG by peripheral inoculation of animal models of inflammatory and neuropathic pain and the analgesic efficacy and side effect profiles will be established following glycine-induced GlyR11-mediated silencing.

描述（由申请人提供）：慢性疼痛是影响 8000 万美国人一生中某个时期的主要健康问题。目前的药物疗法不能长期有效，这导致了基因治疗方法的开发和测试。我们和其他人已经证明，基于 1 型单纯疱疹病毒 (HSV) 的载体可以在接种外周组织后将高效的疼痛调节转基因传递到体内感觉神经元。这种方法的一个主要优点是局部载体递送可以专门针对疼痛组织。我们相信，通过针对负责慢性疼痛的特定神经元类型，可以进一步扩展这一优势，从而使基因转移能够针对特定类型的疼痛（例如炎症或神经性疼痛）进行定制，同时减少有害的副作用。我们最近表明，HSV 介导的甘氨酸受体 11 亚基 (GlyR11) 的转移和长期表达可用于控制外源性施用甘氨酸的传入沉默的时间和持续时间。基于这些发现以及我们最近成功创建高效、完全重定向的 HSV 载体，我们假设我们可以选择性地沉默负责神经性疼痛和炎性疼痛的初级传入神经的不同亚群，从而提供损伤特异性疼痛缓解。这些研究将使我们能够确定相同或不同的传入群体是否是炎症和神经性疼痛的基础，并为未来开发基于 HSV 的基因转移载体提供合理的基础，该载体旨在将镇痛限制于相关的初级传入。我们预计这些研究将（i）提供一种新颖的平台技术，使我们能够选择性地表达旨在调节感觉传入亚群功能的转基因，这种策略可以很容易地扩展到其他类型的感觉神经障碍； (ii) 开发感觉传入亚型的初始功能和物理图，这些亚型在沉默后将阻止不同的持续性疼痛状况，从而提供有针对性的疼痛控制所需的基本信息； (iii) 识别功能改变以响应疼痛刺激的传入神经，提供有关神经纤维可塑性的进一步信息； (iv) 识别与不适当的感觉神经元群体沉默相关的潜在风险（例如本体感觉改变）。为了实现这些目标，我们提出了 3 个具体目标中描述的一系列相互关联的实验。在目标 1 中，HSV 载体的感染性将被重新定位以选择性转导 (a) A2 纤维、(b) 肽能纤维和 (c) 非肽能 C 纤维。在目标 2 中，我们将使用转基因启动子通过转录靶向来补充转导重靶向，当与转导靶向结合时，转基因启动子将微调沉默特异性。组合策略旨在创建较大转导目标群体内感觉纤维亚群的初始精细图，代表对不同疼痛刺激反应的关键传入神经。在目标 3 中，将通过炎症和神经性疼痛动物模型的外周接种将重定向的 HSV 载体引入 DRG，并在甘氨酸诱导的 GlyR11 介导的沉默后建立镇痛功效和副作用曲线。