基于单细胞测序解析APR3通过介导前扣带皮层兴奋性神经元LTP的形成调节神经病理性痛的机制研究

我国约有一亿的慢性痛患者，长期疼痛不仅会影响睡眠、工作和生活，还会增加抑郁、焦虑等的发病率。然而，目前对于慢性痛的细胞和分子调节机制尚不清楚。本课题预实验显示：①单细胞测序发现小鼠前扣带皮层(ACC)多种类型的细胞都参与对神经病理性痛的调节；②生物信息学分析发现兴奋性神经元中凋亡相关蛋白3(APR3)的表达显著下调；③兴奋性神经元APR3敲除小鼠的缩足反应阈值明显降低并伴有自发痛；APR3影响LTP而非LTD的形成，敲除鼠ACC区的自发兴奋性突触后电流的幅值显著增大。这些结果强烈提示APR3在疼痛的调节中起重要作用。本项目拟将进一步研究神经病理性痛状态下ACC区的单细胞特性、不同类型细胞的基因表达差异及细胞间的相互作用；其次明确兴奋性神经元中的APR3在疼痛调节中的作用机理；最后阐明APR3的直接相互作用蛋白及其具体的信号调节通路。本研究将为神经病理性痛的治疗和药物研发提供新思路。

There are about 100 million chronic pain patients in China. Long-term pain will not only affect patients' sleep, work and quality of life, but also increase the incidence of depression, anxiety and other emotional disorders. However, the cellular and molecular mechanisms of chronic pain are currently unknown. In a mouse neuropathic pain model, our preliminary results showed that various types of cells play important roles in the maintenance of pain in anterior cingulate cortex (ACC), and apoptosis-related protein 3 (APR3) was down-regulated in excitatory neurons. Hypersensitivity was induced in CaMKIIα-Cre: APR3flox/flox conditional knockout mice. The formation of LTP but not LTD was affect in ACC of APR3 knockout mice. Moreover, the amplitude of sEPSCs was increased significantly as well, suggesting that APR3 plays a key role in the regulation of pain. We want to research, firstly, the single-cell characteristics, gene expression differences in various types of cells, and cell-to-cell interactions in ACC under neuropathic pain conditions. Then to clarify the role of APR3 in the regulation of pain in excitatory neurons. Finally, to investigate the direct interaction proteins of APR3 in excitatory neurons and their signal pathways. Our study will provide new targets for the treatment and drug development of neuropathic pain.