环状RNAcircTmeff1-Tomoregulin-1调控网络对毒品潜伏心理渴求的影响及其机制研究

Persistent drug craving is an important cause of relapse. As a key regulator, circRNA played an important role in neurological diseases such as Parkinson's disease and depression. However, the effect of circRNA on drug craving is not clear. The results from preliminary study by circRNA-seq techniques and bioformatics analysis indicated that circTmeff1-Tomoregulin-1 network in the nucleus accumbens has significant functions in drug craving. The present project will firstly establish the incubation of drug craving animal model by CPP and SA procedure and then over-express or knockdown circTmeff1 by chemical genetic techniques, which aims to clarify whether circTmeff1 could regulate the Tomoregulin-1 expression and influence the incubation of drug craving process. Furthermore, we will study the circRNA-miRNA-mRNA and circRNA-protein interaction to clarify the mechanism of circTmeff1-Tomoregulin-1 network. The influence of circTmeff1-Tomoregulin-1 network on the incubation of drug craving and underlying mechanisms including the plasticity of AMPA receptor will also be explored. The development of this project will enrich the knowledge of drug addiction and provide new intervention targets for the relapse medication.

持续存在的毒品心理渴求是导致复吸的重要原因。circRNA作为一种重要的调控分子在帕金森、抑郁症等神经系统疾病中发挥重要作用，但在毒品心理渴求中的作用尚未明确。我们前期研究通过circRNA-seq及生物信息学分析发现，伏隔核circTmeff1-Tomoregulin-1调控网络在毒品潜伏心理渴求过程中可能发挥重要作用。本项目拟通过条件性位置偏爱及静脉自身给药建立潜伏心理渴求模型，结合化学遗传学技术过表达或敲减circTmeff1，明确其是否可以调节Tomoregulin-1表达并影响潜伏心理渴求过程；从circRNA-miRNA-mRNA和circRNA-蛋白互作角度研究circTmeff1调节Tomoregulin-1表达的网络机制，并阐明该网络对毒品潜伏心理渴求的影响及谷氨酸受体可塑性机制。项目的开展可丰富对毒品成瘾机制的认识，为复吸防治提供新的干预靶点。

持续存在的毒品心理渴求是导致复吸的重要原因。circRNA作为一种重要的调控分子在神经系统疾病中发挥重要作用，但在毒品心理渴求中的作用尚未明确。本项目首先建立条件线索诱导的潜伏心理渴求模型，通过高通量测序技术发现潜伏心理渴求小鼠伏隔核内circRNA表达谱发生改变；结合多西环素调控AAV技术过表达或调低circTmeff-1，可以明显增强或抑制小鼠潜伏心理渴求；双荧光素酶实验、RNA pulldown、FISH实验证明circTmeff-1可以选择性结合miR-541-5p和miR-6934-3p，生物信息学和western blot分析表明，囊泡相关膜蛋白1 (VAMP1)和神经束蛋白(NFASC)，两者都是miR-541-5p和miR-6934-3p的重叠靶标并在潜伏期间表达增加，构建了circTmeff-1 --miR-541-5p/miR-6934-3p--VAMP1/NFASC调控网络；同时发现伏隔核核部circTmeff-1亲本基因蛋白Tmeff-1在毒品潜伏心理渴求中也发挥重要的调控作用，并且其受到miR-592-3p的负向调节；此外，课题组进一步明确circTmeff-1在可卡因成瘾记忆再巩固的作用及circTmeff-1可能作为miR-206的诱饵调节BDNF的表达的作用机制。本研究明确了circTmeff-1在毒品成瘾记忆再巩固及潜伏心理渴求中的作用及机制，其可成为治疗毒品成瘾，有效防止复吸的潜在治疗靶点，为毒品成瘾治疗提供新思路新理论。