M.I.N.D. Institute, Department of Neurology, University of California at Davis, Sacramento, 95817, USA.
Am J Med Genet B Neuropsychiatr Genet. 2011 Jan;156B(1):72-8. doi: 10.1002/ajmg.b.31140. Epub 2010 Nov 17.
Tourette Syndrome (TS) is diagnosed based upon clinical criteria including motor and vocal tics. We hypothesized that differences in exon expression and splicing might be useful for pathophysiology and diagnosis. To demonstrate exon expression and alternatively spliced gene differences in blood of individuals with TS compared to healthy controls (HC), RNA was isolated from the blood of 26 un-medicated TS subjects and 23 HC. Each sample was run on Affymetrix Human Exon 1.0 ST (HuExon) arrays and on 3' biased U133 Plus 2.0 (HuU133) arrays. To investigate the differentially expressed exons and transcripts, analyses of covariance (ANCOVA) were performed, controlling for age, gender, and batch. Differential alternative splicing patterns between TS and HC were identified using analyses of variance (ANOVA) models in Partek. Three hundred and seventy-six exon probe sets were differentially expressed between TS and HC (raw P < 0.005, fold change >|1.2|) that separated TS and HC subjects using hierarchical clustering and Principal Components Analysis. The probe sets predicted TS compared to HC with a >90% sensitivity and specificity using a 10-fold cross-validation. Ninety genes (transcripts) had differential expression of a single exon (raw P < 0.005) and were predicted to be alternatively spliced (raw P < 0.05) in TS compared to HC. These preliminary findings might provide insight into the pathophysiology of TS and potentially provide prognostic and diagnostic biomarkers. However, the findings are tempered by the small sample size and multiple comparisons and require confirmation using PCR or deep RNA sequencing and a much larger patient population.
妥瑞氏症(TS)是基于运动性和发声性抽动等临床标准进行诊断的。我们假设外显子表达和剪接的差异可能有助于病理生理学和诊断。为了证明与健康对照(HC)相比,TS 患者血液中外显子表达和选择性剪接基因的差异,我们从 26 名未经治疗的 TS 患者和 23 名 HC 的血液中分离出 RNA。每个样本均在 Affymetrix Human Exon 1.0 ST(HuExon)芯片和 3'偏向 U133 Plus 2.0(HuU133)芯片上运行。为了研究差异表达的外显子和转录本,我们采用协方差分析(ANCOVA)进行分析,控制年龄、性别和批次。采用 Partek 中的方差分析(ANOVA)模型来识别 TS 和 HC 之间差异表达的选择性剪接模式。376 个外显子探针集在 TS 和 HC 之间存在差异表达(原始 P<0.005,倍数变化> |1.2|),通过层次聚类和主成分分析可以区分 TS 和 HC 患者。使用 10 倍交叉验证,这些探针集对 TS 患者的预测准确率超过 90%。90 个基因(转录本)在外显子水平存在差异表达(原始 P<0.005),并且与 HC 相比,在 TS 中存在选择性剪接(原始 P<0.05)。这些初步发现可能为妥瑞氏症的病理生理学提供深入的见解,并可能提供预后和诊断生物标志物。但是,由于样本量小且进行了多次比较,这些发现需要使用 PCR 或深度 RNA 测序以及更大的患者群体进行验证。