Sigma 1 receptor as a master regulator of pronociceptive protein trafficking in t

描述（由申请人提供）：由慢性炎症和神经系统损伤引起的神经性疼痛特别难以用目前可用的drg器械治疗，并且是高度成问题的未解决的临床问题。开发一种新的治疗神经性疼痛的方法是非常重要的。许多信号分子可能参与神经性疼痛的表现。σ 1受体（S1 R）是一种主要存在于内质网的双跨膜蛋白，在基本细胞过程中具有多种作用。对S1 R敲除小鼠描述的最稳健的表型之一是神经性疼痛的抑制。鞘内施用S1 R拮抗剂反映了这种敲除的抗神经病表型，表明S1 R在介导神经病性疼痛中的脊髓节段水平表达的重要性。基于S1 R在脊髓腹角的集中表达，我们认为背根神经节（DRG）中的S1 R是其作用的关键解剖部位。我们提出了一个新的概念，即S1 R作为DRG中前感受性蛋白运输的主调节因子，作为在具有该蛋白的药理学或遗传抑制的小鼠中观察到的抗神经病表型的基础机制。初步数据证明S1 R和几种原伤害感受蛋白之间的蛋白质：蛋白质相互作用，包括P物质受体（NK 1 R）、毒蕈碱M1受体（M1 R）和NMDA受体的NR 1亚基，它们都具有增加细胞内钙信号传导的能力。S1 R的共表达增加了这些细胞中NK 1 R的质膜表达，从而增加了细胞内钙信号传导。我们将测试高度创新的工作假设：S1 R蛋白上调和调制的NK 1 R，M1 R，和NMDAR在背根神经节（DRG）介导的神经病理性疼痛。该提案采用了最先进的技术，包括选择性的在体内转导和敲低的S1 R在背根神经节的腺相关病毒2/8，广泛使用免疫共沉淀和新的分子构建体来破译负责S1 R和客户蛋白之间的相互作用的蛋白质结构域，并最终研究了神经性疼痛的基因治疗靶向S1 R与客户蛋白的相互作用。

DESCRIPTION (provided by applicant): Neuropathic pain resulting from chronic inflammation and injury to the nervous system is particularly difficult to treat with the currently available drg armamentarium and is a highly problematic unsolved clinical problem. Development of a novel treatment for neuropathic pain is highly significant. Many signaling molecules are likely to partake in the manifestation of neuropathic pain. The Sigma 1 receptor (S1R) is a two-transmembrane mostly endoplasmic reticulum resident protein with many roles in fundamental cellular processes. One of the most robust phenotypes described for the S1R knockout mice is the suppression of neuropathic pain. Intrathecal administration of S1R antagonists mirrors this anti-neuropathic phenotype of the knockout indicating the importance of a spinal segmental level expression of S1R in mediating neuropathic pain. Based on the focused expression of S1R in the ventral horn in the spinal cord proper, we believe S1R in the dorsal root ganglion (DRG) is the key anatomical site of action. We propose a novel concept of the S1R as a master regulator of pronocicptive protein trafficking in the DRG as a mechanism underlying the anti- neuropathic phenotype observed in mice with pharmacological or genetic inhibition of this protein. Preliminary data documents protein:protein interaction between S1R and several pronociceptive proteins including the substance P receptor (NK1R), muscarinic M1 receptor (M1R), and the NR1 subunit of the NMDA receptor all sharing the ability to increase intracellular calcium signaling. Co-expression of S1R increases the plasma membrane expression of NK1R in these cells with a consequent increase in intracellular calcium signaling. We will test the highly innovative working hypothesis that: S1R protein upregulation and modulation of NK1R, M1R, and NMDAR in the dorsal root ganglion (DRG) mediates neuropathic pain. The proposal incorporates state-of-art techniques including a selective in vivo transduction and knockdown of S1R in the DRG by adenoassociated virus 2/8, extensive use of co-immunoprecipitation and novel molecular constructs to decipher the protein domains responsible for the interaction between S1R and the client proteins, and finally examines a gene therapy for neuropathic pain targeting S1R interaction with client proteins.