塌陷反应调节蛋白CRMP2磷酸化作为慢性痛治疗新靶点的验证及其介导的柚皮素镇痛作用

慢性疼痛（Chronic Pain，CP）影响超过全球35%的人口，严重影响患者生存质量。靶向阻断电压门控钙通道Cav2.2的镇痛药物是各大制药企业研发的热点，但这些药物存在有效剂量过大引起的严重副作用、个体化给药要求高等一系列问题。申请人根据前期大量预实验提出调节胞质塌陷反应调节蛋白2（CRMP2）磷酸化间接影响Cav2.2的表达和功能，抑制CP的全新思路。因此本项目拟利用多种转基因小鼠、转染质粒、磷酸化抑制剂和多种行为学方法，研究CRMP2磷酸化参与CP的作用和特点。通过细胞及脊髓片钙成像、膜片钳，结合在体多通道微电极和离体微阵列等技术阐明CRMP2磷酸化修饰间接影响Cav2.2的表达和功能，及干预中枢敏化的机制。最后评价CRMP2磷酸化调节相关天然产物柚皮素（NAR）的镇痛作用。本项目将为CRMP2磷酸化修饰作为CP治疗新靶点提供更为明确的证据，为靶向药物研发和临床转化提供实验依据。

Chronic Pain (CP) affects more than 35% of the world's population and seriously affects the quality of life of patients. Analgesic drugs that directly block voltage-gated Cav2.2 are hotspots developed by international pharmaceutical companies, but these drugs have serious side effects caused by excessive effective doses, and high requirements for individualized drug delivery. Dependent on a large number of preliminary experiments, we proposed a new idea to indirectly inhibit the expression and function of Cav2.2 by regulating phosphorylation of Collapsin Response Mediator Protein 2 (CRMP2). Therefore, this project intends to use a variety of transgenic mice, transfection plasmids, phosphorylation inhibitors and a variety of behavioral methods to study the role and characteristics of CRMP2 phosphorylation involved in CP. Through cell and spinal cord calcium imaging, patch clamp, combined with multi-channel microelectrodes and ex vivo microarrays, we will investigate the indirectly effects of phosphorylation of CRMP2 on the expression and function of Cav2.2, and the mechanism of intervention in central sensitization. Finally, the analgesic effect of a CRMP2 phosphorylation regulation related natural compound naringenin (NAR) will be evaluated. This project will provide more clear evidence for CRMP2 phosphorylation as a new target for CP therapy, providing experimental evidence for targeted drug development and clinical transformation.