基于全基因组外显子测序的中国汉族海洛因依赖遗传学分析

The genetic variations of exome which play an important role in the complex mental disorders influence the occurrence, development, clinical manifestation and curative effect of heroin dependence. Recently years, some polymorphism analysis about drug dependence has been reported, but the underlying mechanism has not yet been elucidated for the variety of the subjects and methods. In this project, we apply the whole exome sequencing to Han Chinese population of heroin addicts and healthy controls, and then obtain the exome variations map of heroin dependence. Through the association analysis, haplotype analysis, etc, we aim to identify the key exon variations. We explain the pattern of multi-factors’ interactionby performing the gene-gene, gene-environment interaction analysis. Explore the effective intervene strategies and treatment measures that can be obtained by analyzing the association between exome variations and the characters of clinic and epidemiology. The evaluation of the methadone maintenance treatment effect is also one of the important purposes in this study. In order to clarify the biological mechanism of the key exon variation, we investigate the expression level of some key genes. This study can not only provide the experimental evidence for further research on molecular mechanism, but also contribute to the theoretical basis for personalized medicine and the new treatment strategy in heroin dependence.

全基因组外显子序列变异是影响精神疾病发生、发展及临床治疗的关键因素之一，其在海洛因依赖演进过程、临床预防及治疗中具有重要作用。近年来，虽有不少毒品依赖相关基因多态性研究报道，但因研究材料和方法各异，以至于毒品依赖发生、发展机制仍未明了。本项目拟采用全基因组外显子测序技术分析中国汉族海洛因依赖及健康群体全基因组外显子序列，获得全基因组外显子突变谱；利用关联性分析、单倍型分析等确定海洛因依赖易感基因及单倍型；通过基因-基因、基因-环境互作分析，阐明多元素交互作用对海洛因依赖发生、发展的影响；分析相关遗传变异与海洛因依赖临床及流行病学特征的相关性，寻求海洛因依赖的有效干预策略和治疗手段；继而检测关键基因的表达水平，探索相关遗传变异影响海洛因依赖的生物学机制。本课题不仅为深入研究基因变异影响海洛因依赖发生、发展的分子机制提供实验基础，同时还为海洛因依赖的个体化医疗及新药开发提供理论依据。

药物依赖是一种慢性复发性脑病，其给公共卫生、社会治安和经济发展带来严重负担，对药物依赖发生发展机制的深入研究具有重要的经济、社会意义。本研究从遗传变异可显著影响疾病发生发展的角度出发，评估了FAT非典型钙粘蛋白3（FAT atypical cadherin 3，FAT3）、驱动结合蛋白1（kinectin 1，KTN1）、突触后致密蛋白93（discs large homolog 2，DLG2）、结肠癌缺失基因（deleted in colorectal cancer gene，DCC）、5羟色氨受体3B（5-hydroxytryptamine receptor 3B，HTR3B）、强啡肽原（prodynorphin，PDYN）及κ阿片受体（κ-opioid receptor，KOR）基因多态性与海洛因依赖发病风险及临床表现的相关性，结果表明DCC rs16956878、DCC rs12607853、DCC rs2292043、HTR3B rs1176746、HTR3B rs1185027以及PDYN 68bp-VNTR的多态性与中国汉族海洛因依赖的发病风险显著相关，DCC rs16956878与HTR3B rs1176746的多态性与海洛因依赖者的吸毒方式相关，PDYN 68bp-VNTR的多态性与海洛因静脉使用量及药物戒断次数相关；同时，结合当前神经生物学研究热点，本研究采用肠道微生物16s多样性测序技术，探索了海洛因依赖、海洛因戒断、海洛因依赖美沙酮治疗及健康群体的肠道微生物组成，获得了不同海洛因依赖群体的肠道微生物构成图谱，并分析了不同海洛因依赖群体中存在差异分布的微生物群体，结果表明相对于美沙酮治疗患者，海洛因依赖者的肠道微生物多样性较低，且有益菌丰度减少而有害菌丰度增高；此外，结合申请人上一国家自然科学基金项目，本研究还探讨了糖原合成酶激酶3β（glycogen synthase kinase3，GSK3β）在青春期甲基苯丙胺（methamphetamine，METH）暴露所致成年期行为异常中的作用，结果表明抑制GSK3β活性可对青春期METH暴露所致成年期行为异常产生显著的保护作用。本研究结果不仅可以为阿片类药物及METH依赖的神经生物学机制的进一步研究提供科学基础，还可以为其精准治疗及药物开发提供靶点参考。