Fogel Brent L, Cho Ellen, Wahnich Amanda, Gao Fuying, Becherel Olivier J, Wang Xizhe, Fike Francesca, Chen Leslie, Criscuolo Chiara, De Michele Giuseppe, Filla Alessandro, Collins Abigail, Hahn Angelika F, Gatti Richard A, Konopka Genevieve, Perlman Susan, Lavin Martin F, Geschwind Daniel H, Coppola Giovanni
Program in Neurogenetics, Department of Neurology and
Program in Neurogenetics, Department of Neurology and.
Hum Mol Genet. 2014 Sep 15;23(18):4758-69. doi: 10.1093/hmg/ddu190. Epub 2014 Apr 23.
Senataxin, encoded by the SETX gene, contributes to multiple aspects of gene expression, including transcription and RNA processing. Mutations in SETX cause the recessive disorder ataxia with oculomotor apraxia type 2 (AOA2) and a dominant juvenile form of amyotrophic lateral sclerosis (ALS4). To assess the functional role of senataxin in disease, we examined differential gene expression in AOA2 patient fibroblasts, identifying a core set of genes showing altered expression by microarray and RNA-sequencing. To determine whether AOA2 and ALS4 mutations differentially affect gene expression, we overexpressed disease-specific SETX mutations in senataxin-haploinsufficient fibroblasts and observed changes in distinct sets of genes. This implicates mutation-specific alterations of senataxin function in disease pathogenesis and provides a novel example of allelic neurogenetic disorders with differing gene expression profiles. Weighted gene co-expression network analysis (WGCNA) demonstrated these senataxin-associated genes to be involved in both mutation-specific and shared functional gene networks. To assess this in vivo, we performed gene expression analysis on peripheral blood from members of 12 different AOA2 families and identified an AOA2-specific transcriptional signature. WGCNA identified two gene modules highly enriched for this transcriptional signature in the peripheral blood of all AOA2 patients studied. These modules were disease-specific and preserved in patient fibroblasts and in the cerebellum of Setx knockout mice demonstrating conservation across species and cell types, including neurons. These results identify novel genes and cellular pathways related to senataxin function in normal and disease states, and implicate alterations in gene expression as underlying the phenotypic differences between AOA2 and ALS4.
由SETX基因编码的Senataxin蛋白在基因表达的多个方面发挥作用,包括转录和RNA加工。SETX基因突变会导致隐性疾病2型动眼神经失用性共济失调(AOA2)和显性青少年型肌萎缩侧索硬化症(ALS4)。为了评估Senataxin蛋白在疾病中的功能作用,我们检测了AOA2患者成纤维细胞中的差异基因表达,通过微阵列和RNA测序鉴定出一组表达发生改变的核心基因。为了确定AOA2和ALS4突变是否对基因表达有不同影响,我们在Senataxin蛋白单倍体不足的成纤维细胞中过表达疾病特异性的SETX突变,并观察不同基因集的变化。这表明Senataxin蛋白功能的突变特异性改变在疾病发病机制中起作用,并为具有不同基因表达谱的等位基因神经遗传性疾病提供了一个新的例子。加权基因共表达网络分析(WGCNA)表明,这些与Senataxin蛋白相关的基因参与了突变特异性和共享的功能基因网络。为了在体内评估这一点,我们对12个不同AOA2家族成员的外周血进行了基因表达分析,并确定了一个AOA2特异性转录特征。WGCNA在所有研究的AOA2患者外周血中鉴定出两个高度富集该转录特征的基因模块。这些模块是疾病特异性的,并且在患者成纤维细胞和Setx基因敲除小鼠的小脑中得以保留,这表明在包括神经元在内的不同物种和细胞类型中具有保守性。这些结果确定了与正常和疾病状态下Senataxin蛋白功能相关的新基因和细胞途径,并表明基因表达的改变是AOA2和ALS4之间表型差异的基础。