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，用现有的药物治疗特别困难，是一个高度棘手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。Sigma 1受体（S1R）是一种双跨膜内质网驻留蛋白，在基本细胞过程中起着许多作用。S1R基因敲除小鼠最强大的表型之一是神经性疼痛的抑制。鞘内给予S1R拮抗剂反映了这种基因敲除的抗神经性表型，这表明S1R在脊髓节段水平表达在介导神经性疼痛中的重要性。基于S1R在脊髓内腹角的集中表达，我们认为S1R在背根神经节（DRG）是起作用的关键解剖部位。我们提出了一个新的概念，即S1R作为DRG中预见性蛋白运输的主要调节因子，是在药物或遗传抑制该蛋白的小鼠中观察到的抗神经病变表型的机制。初步数据记录蛋白质：S1R与包括P物质受体（NK1R）、毒毒碱M1受体（M1R）和NMDA受体NR1亚基在内的几种前感觉蛋白之间的蛋白质相互作用都具有增加细胞内钙信号传导的能力。S1R的共表达增加了这些细胞中NK1R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：S1R蛋白上调和NK1R、M1R和NMDAR在背根神经节（DRG）中的调节介导神经性疼痛。该提案结合了最先进的技术，包括通过腺相关病毒2/8在DRG中选择性转导和敲低S1R，广泛使用共免疫沉淀和新型分子构建来解码负责S1R与客户蛋白之间相互作用的蛋白质结构域，并最终研究了针对S1R与客户蛋白相互作用的神经性疼痛的基因治疗。

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.