The schizophrenia candidate gene MIR137: functional studies in mouse

描述（由申请人提供）：精神分裂症（SCZ）是一种复杂且往往具有毁灭性的疾病，终生患病率为0.4%7，遗传率为~80% 8它被广泛认为是神经发育的起源。9,10目前有强有力的遗传学证据表明，MIR137基因及其miRNA产物miR-137与SCZ的病因有关。与该地区的关联现在达到1.7x10-12 （N=59,318）。有14个基因预测或证实了miR-137的靶点，在小GWAS p值下，预测miR-137靶点的基因显著富集（p<0.01）。此外，miR-137还与神经发育、阿尔茨海默病2、智力残疾3、亨廷顿舞蹈病4和Rett综合征小鼠模型有关因此，miR-137是一种具有重要生物学功能的miRNA，也是SCZ和其他神经系统疾病的有趣候选者。因此，我们建议验证miR-137对小鼠神经元发育很重要的假设，确定miR-137调节的途径，并测试这些途径是否在SCZ的发病机制中发挥作用。使用Mir137基因被删除的小鼠，我们将首先描述神经或行为缺陷的小鼠。为了解决miR-137在神经元细胞增殖和分化中的作用，我们将评估脑的大小和结构、神经元密度、神经发生和树突复杂性。已知mirna通过靶向降解或翻译抑制直接调节其他rna。因此，我们将使用RNAseq和PAR-CLIP（一种基于免疫沉淀的方法，用于识别miRNA调节复合体结合的rna）的创新组合来识别小鼠脑中miR-137的直接和间接靶标。我们将分析这些数据，以确定受miR-137影响的单个基因和生物学途径或网络。最后，我们将通过比较现有SCZ GWAS （PGC， bbb31000例）、外显子组测序（瑞典，2500例）和死后基因表达研究的结果，来解决miR-137调控的通路和网络是否在SCZ中被破坏。这些实验将评估miR-137在小鼠大脑中的作用，确定miR-137的潜在靶点（包括原发性和继发性），并测试miR-137控制的通路在SCZ中的失调。这些结果可以为支持SCZ的高度显著的共同危险因素提供功能证据。

DESCRIPTION (provided by applicant): Schizophrenia (SCZ) is a complex and often devastating disease with a lifetime prevalence of 0.4%7 and heritability of ~80%.8 It is widely believed to be of neurodevelopmental origin.9, 10 There is now strong genetic evidence implicating the gene MIR137 and its microRNA (miRNA) product miR-137 in the etiology of SCZ. Association with this region has now reached 1.7x10-12 (N=59,318).1 14 genes have predicted or confirmed miR-137 target sites and genes with predicted miR-137 targets were significantly enriched for small GWAS p- values (p<0.01). In addition, miR-137 is implicated in neurodevelopment, Alzheimer's disease2, intellectual disability3, Huntington's disease,4 and mouse models of Rett syndrome.5 Therefore, miR-137 is a miRNA of important biological function and an intriguing candidate in SCZ and other neurological disease. We therefore propose to test the hypothesis that miR-137 is important for neuronal development in the mouse, identify pathways regulated by miR-137, and test whether these pathways play a role in the pathogenesis of SCZ. Using mice in which the Mir137 gene has been deleted, we will first characterize the mice for neurological or behavioral deficits. To address the role of miR-137 in proliferation and differentiation of neuronal cells, we will evaluate brain size and structure, neuronal density, neurogenesis, and dendritic complexity. miRNAs are known to directly regulate other RNAs through targeting for degradation or translational inhibition. Therefore, we will use an innovative combination of RNAseq and PAR-CLIP (an immunoprecipitation based method to identify RNAs bound by the miRNA regulation complex) to identify both direct and indirect targets of miR-137 in mouse brain. We will analyze these data to identify individual genes and biological pathways or networks influenced by miR-137. Finally, we will address whether the pathways and networks regulated by miR-137 are disrupted in SCZ by comparing to results from existing SCZ GWAS (PGC, >31,000 cases), exome sequencing (Sweden, 2500 cases), and post-mortem gene expression studies. These experiments will evaluate the role of miR-137 in the mouse brain, identify potential targets of miR-137 (both primary and secondary), and test for dysregulation of the miR-137 controlled pathway in SCZ. These results could provide functional evidence in support of a highly significant common risk factor for SCZ.