基于microRNA技术的精神分裂症相关生物标记物的遗传流行病学研究

Schizophrenia is one of the most serious mental disorder and a kind of complex disease, its pathogenesis and etiology are not yet fully elucidated，and its objective diagnostic biomarkers is lack. On the basis of this study in early about of schizophrenia-specific expression protein and candidate gene in coding region, it would be researched that to find and verify that the Chinese group's plasma biomarkers of microRNAs. Bioinformatics techniques was applicated to predict miRNAs that regulate schizophrenia-specific expression protein. Deep sequencing technology was used to screen differentially expression in plasma microRNAs. To predict differentially expressed microRNAs in GEO database by bioinformatics technology. Through comparing the level of candidate miRNAs that was detected by TaqMan® RT-PCR in Northern Han Chinese schizophrenia patients group and normal subjects group, it would be confirmed that the relationship between miRNAs and schizophrenia. At the same time, we predicted SNPs in the sequence that transcription of miRNA genes, then detected SNPs polymorphism in a large sample of schizophrenia family trios by sequenom MassArray time-of-flight mass spectrometry genotyping technique. The positive SNPs would be validation with a large of case-control samples. Through comparing SNPs polymorphism in case group and control group, the association of candidate gene loci and schizophrenia would be confirmed. In this study, gene expression regulation was researched in two levels of miRNA levels and miRNA gene sequence level, to analysis of the regulation of disease-specific protein expression of miRNAs and schizophrenia. It may provide important clues to clarify the molecular mechanisms of schizophrenia and also may be a new role for the corresponding drug development target and biomarkers of diagnosis and prognosis.

精神分裂症是最严重的常见精神疾病之一，其发病机制迄今未明，且缺乏客观诊断生物标志物。本研究采用miRNA深度测序筛选、调控候选基因miRNA检测与生物信息学预测相结合的方法，寻找并验证中国人群精分症血液miRNA生物标志物。以中国北方汉族精分症病人和正常人为研究对象，采用深度测序技术筛选血浆中差异表达的miRNA，结合生物信息学技术预测GEO数据库差异表达的miRNA以及调控特异表达蛋白的miRNAs，应用RT-PCR技术检测大样本病例对照血液样本中候选miRNAs的水平，证实miRNAs与精分症的关系。预测转录miRNA基因序列中的SNPs，检测精分症核心家系SNPs多态性，并用大样本病例对照样本验证阳性位点，分析与miRNA相关基因位点与精分症的关系。本研究从miRNA水平和miRNA基因序列水平分析验证精分症的miRNAs生物标志物，为阐明精分症发病分子机制及诊断试验的开发提供依据。

精神分裂症是最严重的常见精神疾病之一，其发病机制迄今未明，且缺乏客观诊断生物标志物。本研究采用miRNA深度测序筛选、调控候选基因miRNA检测与生物信息学预测相结合的方法，寻找并验证中国人群精分症血液miRNA生物标志物。以中国北方汉族精分症病人和正常人为研究对象，采用深度测序技术筛选血浆中差异表达的miRNA，结合生物信息学技术预测GEO数据库差异表达的miRNA以及调控特异表达蛋白的miRNAs，应用RT-PCR技术检测大样本病例对照血液样本中候选miRNAs的水平，证实miRNAs与精分症的关系。预测转录miRNA基因序列中的SNPs，检测精分症核心家系SNPs多态性，并用大样本病例对照样本验证阳性位点，分析与miRNA相关基因位点与精分症的关系。.从基因水平、miRNA水平和蛋白质水平，证实调控纤维蛋白肽A、激肽原L和补体C3f蛋白的miRNA在精神分裂症的发病机制中的重要作用。从遗传和表观遗传机制综合分析候选miRNA与精神分裂症的关系，筛选并验证了miR-9-5p、miR-30a-5p、miR-30e-3p、miR-4467和miR-1908-5p五个miRNA与精神分裂症相关。 CACNA2D2基因rs45536634和rs3806706位点多态性均与精神分裂症相关联，rs45536634位点上携带A等位基因或者rs3806706位点上携带GC基因型均会增加精神分裂症的患病风险。CMIP基因rs2287112位点多态性与精神分裂症相关联，携带GG基因型增加患精神分裂症的风险。miR-499a-3p/hsa-miR-499b-5p的多态性位点rs3746444和miR-323b的多态性位点 rs56103835与中国北方汉族人群精神分裂症的遗传易感性相关。.本研究从miRNA水平和miRNA基因序列水平分析验证精分症的miRNAs生物标志物，为精神分裂症早期诊断、开发新型治疗药物和预测个体发病风险提供理论依据。